All abstracts are listed here (both oral and poster
presentations) by the alphabetical order of the surnames of the first author.
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Aldape, Kenneth National Cancer Institute The NCI/Bethesda DNA Methylation Classifier: an
Important Diagnostic Tool for CNS Tumors DNA methylation-based classification has become
essential for accurately diagnosing CNS tumor subtypes, by providing a
reproducible and objective tool in addition to existing diagnostic
modalities. While the commonly used Heidelberg classifier is highly accurate
and valuable, we find utility in creating an alternative classifier to 1)
provide a ‘second look’ at the methylation results in addition to the
Heidelberg classifier; and 2) offer the flexibility to add recently described
classes to the classifier. To this end, the NCI/Bethesda classifier version
3.0 was developed, starting with a set of 19,642 samples, representing 201
unique tumor types/subtypes. We then split this reference set into an 80%
training set (n=15,467) and a 20% internal validation set (n=3,800). The
15,467-sample training set served as the foundation for building hierarchical
Support Vector Machine (SVM) classifiers. In the first stage, we classified
samples into 22 major tumor superfamilies. In the second layer of classification,
we further subclassified each superfamily into specific tumor types/subtypes.
We compared the results of this classifier with existing sample labels that
had been derived from a previous version of the NCI/Bethesda classifier, as
well as from the Heidelberg version 12.8 classifier. At the individual class
level, mean accuracies ranged between 97% and 99%. Our first level of model
performance evaluation was based on cross-validation accuracies, and we
extended this to predict superfamily and class for the 20% internal
validation set(n=3800), showing 95.6% accuracy (3,633 correctly classified
out of 3,800 samples), using a 0.9 confidence score threshold. We then
applied both the Heidelberg classifier and the NCI/Bethesda v3.0 classifier
to an external validation set n=1963 samples), which was completely unseen at
the time of model creation. With the 0.90 threshold cutoff, the DKFZ
classifier correctly classified only 1,033 out of 1,963 samples, achieving a
match rate of 52.6%. In contrast, the NCI/Bethesda classifier correctly
classifying 1,383 out of the 1,963 samples, achieving a higher match rate of
70.5%. High-confidence matches were reached in both classifiers for 836/1,963
samples. A high-confidence match to only the Heidlberg
classifier was reached in 197 samples, while 547 samples showed a
high-confidence match to only the NCI/Bethesda classifier, highlighting the
potential utility of using both classifiers in daily practice. Two tumor
types in the current WHO CNS classification, absent from the Heidelberg
classifier (Desmoplastic myxoid tumor of the pineal region, SMARCB1-mutant,
and Intracranial mesenchymal tumor, FET::CREB fusion-positive) are present in
the NCI/Bethesda classifier, enabling their identification in suspected
cases. In addition, newly described classes, including Dural angioleiomyoma
and High‑grade glioma with pleomorphic and pseudopapillary features, as
well as others, are included in the NCI/Bethesda classifier. Use of our
classifier is freely available following simple registration at
https://methylscape.ccr.cancer.gov/. In sum, the NCI/Bethesda provides a tool
that can be used to robustly classify CNS tumors, providing an alternative
machine learning method as well as additional tumor types relative to the
Heidelberg classifier. Our current consultation practice employs both
classifiers, in an effort to fully utilize the merits of each. |
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Andriani, Rini Dharmais National Cancer Center of Indonesia Association of Isocitrate Dehydrogenase (IDH) Mutation Status with Seizure Incidence in Glioma Patients: Insights from the Indonesian Cancer Center in 2023-2024 Background: Seizures are a common clinical manifestation in glioma patients and, if uncontrolled, can significantly contribute to morbidity, cognitive decline, and a diminished quality of life. Previous studies have suggested an association between lower-grade gliomas and a higher incidence of seizures, but the underlying mechanisms remain unclear. Objective: This study aims to identify the roles of isocitrate dehydrogenase (IDH) biomarker mutations for clinical seizure prediction in glioma patients in Indonesia. Methods: The study design is retrospective based on the data collected at Dharmais National Cancer Center Hospital, Jakarta, Indonesia. We examined all glioma patients admitted to our hospital between January 1, 2023, and October 1, 2024. All glioma patients were confirmed by histopathological analysis. We examined the association between IDH mutation status and seizure incidence during hospitalization or admission. We collected data on the demographic and clinical characteristics of patients on age, sex, glioma type (astrocytoma or oligodendroglioma), WHO grade (2-4), tumor location (supratentorial or infratentorial), and treatment history. Statistical analyses were performed using Stata version 17.0 to estimate the odds ratio (OR) to represent the degree of association between seizure and IDH using logistic regression. Multivariable logistic regression was used to control for potential confounders. Result: Seizures were observed in 42 of the 66 patients (63.6%); among these, 27 patients (64.3%) had the IDH R132H mutation with a statistically significant association (p < 0.05). Glioma patients with mutated IDH R132H had 3.58 higher odds of experiencing seizures than those with the IDH wild-type (OR = 3.58, 95% CI: 1.07—12.0, p < 0.05), after adjusting to sex and age variable. All patients with oligodendroglioma experienced seizures (p = 0.05). Patients over 50 years old had a lower risk of seizure occurrence compared to patients under 40 (OR = 0.22, 95% CI: 0.063 — 0.76, p < 0.05), while no statistically significant difference was observed for the 40-50 age group (OR = 2.70, 95% CI: 0.39-- 18.8, p > 0.05). Sex was not a statistically significant predictor of seizure risk in glioma patients (male vs. female OR = 2.20, 95% CI: 0.66 —7.36, p > 0.05). Analysis of tumor grade showed no statistically significant difference in seizure incidence among WHO tumor grades 2, 3, and 4. Grade 3 tumors had a higher but non-significant OR for seizures compared to Grade 2 (OR = 3.85, 95% CI: 0.93—15.86, p > 0.05). Conclusion: IDH mutation status is a significant predictor of seizure incidence in glioma patients, compared to wild-type IDH. The age over 50 has a lower risk of seizures than the age below 40, while the age group 40-50 does not demonstrate significant predictive power. These findings highlight the importance of the clinical prediction roles of IDH mutations and younger age in the treatment of glioma-related epilepsy. Keywords: Indonesia, glioma, seizures, isocitrate dehydrogenase (IDH), epilepsy |
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Chang, Qing Beijing Neurosurgical Institute, Bejing Tiantan Hospital Tumor associated macrophage derived exosomes modulate immunotherapeutic sensitivity of SHH medulloblastoma by m6A modification Background: Medulloblastoma (MB) is the most common malignant brain tumor in children. The infiltration of tumor-associated macrophages (TAMs) in tumor microenvironment (TME) is correlated with the poor prognosis in SHH subtype of MB patients. Epigenetic alterations on RNA m6A modification were identified in this tumor. However, it remains unclear whether the TAMs infiltration in TME was correlated with the m6A modification status during tumor progression. Methods: The expression of m6A modification-related proteins and TAMs specific marker were assessed by immunohistochemistry in 40 SHH MB primary tumors. Methylated RNA immunoprecipitation sequencing (MeRIP-seq) and transcriptomic RNA sequencing (RNA-seq) were used to screen target genes affected by TAM-derived exosomes in SHH MB cells. Mechanisms of m6A-modification on target gene (FOXD1) were demonstrated by WB, qPCR, RNA half-life time and RNA immunoprecipitation assays. Combination treatment with AAV2/9-shFOXD1 and PD-1 inhibitors were performed in the NeuroD2: SmoA1 mouse model. Results: We reported for the first time that significant infiltration of TAMs was associated with decreased expression of METTL14 in primary SHH MBs. Exosomal microRNAs (miR-320e, miR-196b-5p, and miR-628-5p) released by TAMs targeted to the 3’-UTR (3’ Untranslated Region) of METTL14 in MB cells, promoting the gene’s degradation and consequently downregulating the overall m6A level in SHH MB. MeRIP-seq and RNA-seq analyses screened out Forkhead box D1 (FOXD1) as the target gene upregulated by TAMs-derived exosomes through m6A modification in MB cells, which was correlated with poor prognosis in SHH MB patients. Further experiments in vitro verified YTHDF2 can recognize m6A-modified FOXD1 and promote its mRNA degradation. Knockdown of FOXD1 in SHH MB cells significantly promoted the release of the CD8+T cell recruiting chemokines CXCL10/11. Treatment with AAV2/9-shFOXD1 significantly enhanced the anti-tumor effect of PD-1 inhibitor in NeuroD2: SmoA1 SHH MB mice. Conclusion: Our study revealed the m6A modification status of SHH MBs influenced by TAMs derived exosomal miRNAs. FOXD1 promoted the malignant progression of SHH MB through m6A modification. Increased immunotherapeutic sensitivity of the combination treatment with FOXD1 knockdown and anti-PD-1 blockade in vivo shed lights on a novel therapeutic strategy of SHH MB. |
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Chen, Bo The University of Hong Kong SIRT5 Suppresses Tumor Growth by Regulating
Mitochondrial Metabolism and Synaptic Remodeling in Gliomas Sirtuin 5 (SIRT5) is increasingly recognized as a
key regulator of cellular metabolism, which is commonly dysregulated in
cancer cells, resulting in enhanced proliferation and tumor progression. To
investigate the clinicopathologic implications of SIRT5 dysregulation in
glioblastoma, we performed comprehensive analyses of transcriptomic data and
functional verifications using in vitro and in vivo glioblastoma models. We
found that higher SIRT5 expression levels were associated with a favorable
prognosis in glioma patients. Knockdown of SIRT5 significantly enhanced
glioblastoma cell growth. Our data suggest its potential role in regulating
mitochondrial metabolism in gliomas. Furthermore, SIRT5 is also significantly
correlated with synaptic remodeling pathways. Our findings indicate a
tumor-suppressive role for SIRT5 that extends beyond regulating cancer
metabolism by which it may function through modulating neuroplasticity.
Understanding these cellular interactions provides nuanced insights into the
multifaceted role of SIRT5 and the broader therapeutic implications for the
development of novel treatment strategies. |
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Chen, Zhilin The University of Hong Kong Single-nucleus and spatial transcriptomics of
intracranial germinoma Intracranial germinoma is the most common pediatric CNS
germ cell tumour. While the survival rates of
patients following standard treatment with radiotherapy plus carbo-etoposide
are high, the children inevitably suffer from long-term cognitive impairment and
neurological damage due to craniospinal irradiation. Germinomas often exhibit
substantial lymphocyte infiltration; however, as they lack expression of
immune checkpoint proteins, making them unlikely candidates for PD-1 or
CTLA-4 blockage therapy. Indeed, the roles of the immune infiltrates in the tumour microenvironment of germinoma are poorly
understood.
We hope that understanding the immune-tumour interactions will lead to the development of less
cytotoxic treatments for patients with germinoma. This could potentially
de-escalate or supplant craniospinal irradiation, enhancing the quality of
life for the patients, while preserving survival outcomes. |
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Chong, Kyuha Samsung Medical Center The Role of Extra-Domain B
Fibronectin in Glioblastoma Cell Invasion Glioblastoma (GBM) is a highly
aggressive tumor marked by its resistance to current treatments and high
recurrence rates. Extra-domain B Fibronectin (EDB-FN), a glycoprotein absent
in normal tissues but selectively expressed in the tumor microenvironment,
was previously identified as a promising molecular marker for brain tumors,
with potential applications in prognostics and therapeutic targeting. Despite
this, the specific role and mechanism of EDB-FN in GBM cells remain
undefined. In this study, the role and mechanisms of EDB-FN in contributing
to GBM malignancy were investigated through in vitro experiments. The results
demonstrate that silencing EDB-FN in GBM cells significantly reduces cell
migration, invasiveness, extracellular matrix adhesion, and stem cell-like
properties. Moreover, it was identified that the FAK and ERK signaling
pathways are crucial for GBM cell progression in response to EDB-FN. This
study highlights the potential of EDB-FN as a specific biomarker for GBM
treatment, paving the way for future research in this area. |
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Clautrier, Simon GH Saintes-Saint Jean d'Angely Potentiation of treatments for
glioblastoma: Analysis of a repositioning strategy for Metformin, Simvastatin
and Valproic Acid associated with the Stupp protocol Glioblastoma (GBM) is the most
aggressive primary glial brain tumor in adults. Despite the Stupp protocol,
prognosis remains poor, with overall survival of 14.6 months. The mechanisms
of resistance are multifactorial: the importance of epigenetic modulations,
the persistence of stem cells and the adoption by the cell of an aerobic
glycolytic metabolism (Warburg effect) are elements that have been considered
to be major causes of radiochemical resistance and short/medium-term failure
of standard treatment. This study explores the potential
repositioning of Valproic Acid (VPA), Metformin, and Simvastatin in GBM
treatment, evaluating their ability to target resistance mechanisms and
potentiate Temozolomide's (TMZ) effect. Material and Method: A review of the literature analysed the mechanisms of action of these molecules on
the radiochemical resistance mechanisms described Results: An analysis of the literature
reveals that : On GBM epigenetic modulations: VPA, as a histone deacetylase
inhibitor (HDACi), modifies gene expression
patterns. VPA decreases O6-methylguanine-DNA methyltransferase (MGMT)
expression and induces methylation of the MGMT promoter, potentially resensitizing TMZ-resistant GBM cells MicroRNA-21 (miR-21) is
overexpressed in 44-100% of GBM, contributing to chemoresistance. miR-21 is
remarkably down-regulated by Metformin in a dose- and time-dependent manner
in certain cancer cell lines found in the literature On GBM cellular metabolism: w
Metformin and
VPA target the Warburg effect by activating AMP-activated protein kinase
(AMPK), reducing tumor growth and promoting apoptosis. On GBM signaling pathways: w
Liver Kinase
B1 (LKB1)/AMPK/mammalian Target Of Rapamycin (mTOR)
pathway: Metformin activates AMPK, inhibiting mTOR. w
HIPPO/Yes-Associated
Protein (YAP)/Transcriptional co-Activator with PDZ-binding motif (TAZ)
pathway: Simvastatin reduces YAP protein activity by inhibiting mevalonate
synthesis. w
FOS Like 1,
AP-1 Transcription Factor Subunit (FOSL1): VPA could indirectly inhibit FOSL1
by increasing miR-138 expression. On cancer stem cells: All three molecules show potential
for targeting GBM stem cells (GSCs) by modulating stem cell markers and
signaling pathways. On tumor suppressor gene p53: w
Metformin and
VPA could impact p53 reactivation, with potentially superior efficacy on
cells with functional p53. VPA/Metformin: synergistic p53-dependent action w
Simvastatin:
preferential action on mutated p53 w
The triple
combination could be relevant regardless of tumour
heterogeneity The favourable physicochemical
properties (liposolubility), low cost and good
general tolerance of these compounds are additional advantages. Discussion/Conclusion: The combination of VPA,
Simvastatin, and Metformin offers a multi-targeted approach to GBM treatment
by: w
Reversing
epigenetic alterations, including MGMT methylation, and modulating microRNA
expression. w
Disrupting
cancer cell metabolism through the Warburg effect. w
Interfering
with key signaling pathways involved in tumor growth and survival. w
Targeting GBM
stem cells. w
Potentially
reactivating tumor suppressor functions, particularly p53. This strategy could address the
limitations of single-molecule approaches in the face of GBM's heterogeneity.
The synergistic effects of these drugs, combined with their ability to
potentiate TMZ treatment, present a promising avenue for improving GBM
therapy. The study suggests that this
combination could improve the efficacy of standard treatment and potentially
overcome resistance mechanisms. However, further preclinical
research is required to elucidate the interactions between these molecules
and validate their combined effects on GBM cells before proceeding with
clinical trials. |
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Cui, Xiaoteng Tianjin Medical University General
Hospital The basic and clinical research of
targeting EGFR and mevalonate pathways synergize the antitumor effect of
temozolomide in glioblastoma Background: Glioblastoma (GBM) represents the
most malignant primary tumor in the central nervous system. GBM patients
endure rapid progression and poor prognosis, with a median survival time of
15 months, despite surgical resection combined with radio- and
temozolomide-based chemo-therapy since the first diagnosis. Metabolic
reprogramming is a remarkable hallmark of GBM, not only producing enough
energy for highly proliferative tumor cells, but also contributing to
immunosuppressive tumor microenvironment formation. Epidermal growth factor
receptor (EGFR) amplification and EGFR-vIII
constitutively activated mutation are often detected in GBM cells, playing
vital roles in the malignant behavior. Emerging evidences have suggested that
hyper-activated EGFR pathway promotes to lipogenesis and cholesterol uptake.
However, whether the EGFR pathway contributes to fatty acid metabolism and
energy generation remains mystery. Material and Methods: Single-cell RNA sequencing, bulk
RNA sequencing, and untargeted metabolomics analysis of fresh GBM specimens
were performed. Mitochondrial function was detected by seahorse XF analyzer
in GBMs with inhibiting EGFR pathway transductions. Orthotopic GBM models
were constructed in immunodeficient nude mice and immunocompetent C57BL/6J
mice. Results: GBMs with high expression of EGFR
had characteristics of lipid remodeling and maintaining high cholesterol
levels. A metabolism-associated RTK-fatty acid-gene signature (RFA) was
well-constructed to predict the prognosis of GBM patients. Inhibition of the
EGFR pathway and mevalonate pathways could remodel energy metabolism by
repressing the tricarboxylic acid cycle and modulating ATP production.
Mechanistically, hyper-activated EGFR pathway promoted the expressions of
ACSS3, ACSL3, and ELOVL2 in an NF-κB-dependent
manner. Moreover, blockade of mevalonate pathway reduced the EGFR level on
the cell membranes, thereby affecting the signal transduction of the EGFR/AKT
pathway. Targeting EGFR pathway and mevalonate pathways enhanced the
antitumor effect of temozolomide in vitro and in vivo. Furthermore, GBM
patients with the treatment of atorvastatin and temozolomide showed a
prolonged progression free survival (PFS) time and overall survival (OS)
time. Conclusion: This study uncovered a regulatory
mechanism by which EGFR pathway activation promotes energy metabolism by
upregulating ACSS3, ACSL3, and ELOVL2 expressions in GBMs. The combinatorial
therapeutic strategy of temozolomide, auxiliary EGFR-TKI, and statins can
benefit GBM patients with high RFA scores by significantly prolonging their
survival time. Now the phase II clinical trials of temozolomide and
atorvastatin against GBM is in the recruiting status (NCT06327451). |
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Dong, Jun The Second Affiliated Hospital of
Soochow University Repurposing of amlodipine against
glioma stem cells through regulating CD133–EGFR/AKT/mTOR cascade to
downregulate SOX2 expression Background: Glioblastoma is the most aggressive
and lethal primary brain tumor in adults with poor prognosis. Previous
studies attribute high therapeutic resistance and short-term recurrence due
to existence of glioma stem cells (GSCs), a valuable target for specific
effective therapy against glioblastoma. Amlodipine is a classical calcium
channel blocker (CCB) with exact anti-tumor effect independent of CCB
activity，which has
potential of repurposing against glioblastoma. Methods: Cell viability, proliferation,
self-renewal, invasive, apoptosis of GSCs were evaluated with CCK-8, EdU incorporation, tumor sphere formation assay, 3D
spheroid invasion assay after administration of amlodipine. Flow cytometry
was applied to detect apoptosis of GSCs, and Western blot assay was performed
to explore the relevant protein levels involved in molecular mechanisms of
amlodipine against GSCs. The interrelationship between GSC markers and key
proteins of EGFR/Akt/mTOR signaling pathway was investigated by
bioinformatics assay with GEPIA and further verified by Western blot.
Exogenous activation of AKT phosphorylation by adding AKT agonist SC79 to
evaluate the role of Akt signaling in the regulation of glioma stemness. The
effect of amlodipine against GSCs was verified in intracranial orthotopic
GSCs-derived tumor model, bioluminescence imaging was applied to monitor
tumors growth in vivo, Ki67 staining was performed to detect proliferation of
GSCs in vivo, TUNEL staining was used to detect apoptosis and expression of
GSC markers in situ was identified by western blot and immunohistochemistry. Results: The present study disclosed that
amlodipine inhibited cell viability, suppressed self-renewal, down-regulated
stemness, induced apoptosis of GSCs obviously, and significantly reduced
GSCs-derived intracranial tumor growth, prolonged survival of tumor-bearing
mice as well. Mechanistically, amlodipine decreased expression of
GSC-specific marker CD133. CD133 deletion destabilized epidermal growth
factor receptor (EGFR) by enhancing EGFR internalization, thereby suppressing
EGFR/Akt/mTOR signaling, leading to down-regulation of core transcriptional
regulator SOX2 expression, and exogenous activation of AKT phosphorylation by
adding AKT agonist SC79 partially reversed the antitumor effects of
amlodipine. Besides, other dihydropyridine CCBs nicardipine and nifedipine as
well as Ca2+ chelator BAPTA-AM did not reveal inhibitory effect against GSCs,
L-type Ca2 + channel agonist also did not attenuate the inhibitory effect of
amlodipine on GSCs, implying existence of the exact effect of amlodipine
against GSCs. Conclusions: Amlodipine suppressed
GSCs-initiated tumor development via regulating CD133 expression to inhibit
EGFR/Akt/mTOR cascade to downregulate SOX2, indicating amlodipine had novel
potential targeting on GSCs against glioblastoma. |
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Dong, Yijun Peking Union Medical College Single cell analysis of multifocal
GBM reveals the function of ferroptosis induced NUPR1+ BMDM in promoting
natural evolution Glioblastoma (GBM) is an aggressive
brain tumor with high levels of heterogeneity, contributing to its resistance
to treatment and poor prognosis. Multifocal GBM, characterized by the
presence of two or more distinct tumor foci within the same patient, presents
a unique model for studying parallel tumor evolution. In this study, we
investigated the role of bone marrow-derived macrophages (BMDMs) in the
natural evolution of multifocal GBM using single-cell RNA sequencing (scRNA-seq), single-cell ATAC sequencing (scATAC-seq) and spatial transcriptomics from multiple
patient samples. We collected data from four
patients with multifocal GBM, analyzing both older and younger lesions. By
comparing copy number variation (CNV) and performing pseudotime
trajectory analysis, we established two distinct evolutionary trajectories,
defining the older lesions as highly evolved and the younger lesions as less
evolved. Tumor subtypes, including mesenchymal (MES)-like tumor cells, were
found to increase in the highly evolved lesions, which were enriched with a
specific subpopulation of BMDMs expressing NUPR1. These NUPR1+ BMDMs
localized predominantly to necrotic regions of the tumor, areas known to
harbor hypoxia and cell death, and were associated with poor patient
prognosis. Further analysis revealed that the
NUPR1+ BMDM subpopulation was related to ferroptosis, a form of regulated
cell death driven by iron-dependent lipid peroxidation. The NUPR1+ BMDMs
exhibited elevated ferroptosis-related gene expression, indicating that they
may play a role in promoting this form of cell death within the tumor
microenvironment. Spatial transcriptomic analysis also showed that SPP1
signaling between MES tumor cells and NUPR1+ BMDMs was enriched in necrotic
regions, suggesting a crosstalk mechanism that drives tumor progression. To validate these findings, we
conducted in vitro experiments using the THP-1 macrophage cell line. We
knocked down NUPR1 expression in these cells using siRNA and observed a
significant decrease in the proliferation and migration of co-cultured GBM
cells. Additionally, the expression of MES markers in the tumor cells was
reduced, indicating that NUPR1+ BMDMs may promote the mesenchymal-epithelial
transition (MET) phenotype, which is linked to increased malignancy in GBM. Overall, our results suggest that
NUPR1+ BMDMs contribute to the natural evolution of GBM by facilitating
ferroptosis and promoting mesenchymal transition, particularly in necrotic
areas. This subpopulation of macrophages represents a potential target for
therapeutic intervention, as disrupting their function could inhibit tumor
progression and improve patient outcomes. Future studies will further explore
the mechanisms of NUPR1 in ferroptosis and its interactions with other immune
and tumor cell populations in the GBM microenvironment. |
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Duan, Zejun Sanbo Brain Hospital, Capital Medical University, Beijing Rare DICER1-mutant primary
intracranial sarcoma: five cases report and review of literatures Objective: To investigate the clinicopathological
characteristics and differential diagnosis of DICER1-mutant primary intracranial
sarcoma. Methods: Five cases of DICER1-mutant primary
intracranial sarcoma at Sanbo Brain Hospital,
Capital Medical University during May 2013 to December 2024 were collected.
The clinical and imaging data were retrieved. H&E and immunohistochemical
staining, as well as next-generation sequencing, were performed.
Additionally, a literature review was conducted. Results All five sarcomas
were located in the supratentorial regions, with one case involving the basal
ganglia, including three females and two males. The median age at diagnosis
was 25 years (range: 8-33 years). Morphologically, they were characterized by
malignant spindle cell sarcoma, with high mitotic activity and cytoplasmic
eosinophilic globules. Myxoid degeneration, necrosis, and invasion into
surrounding brain tissue were observed in some cases. Immunophenotypically,
tumor cells showed diffuse expression of Vimentin and variable expression of myogenic
marker (Desmin), with or without focal MyoD1 and/or
Mogenin. Four cases of tumors exhibited diffuse
strong positive expression of TLE1 and P53. Three cases displayed loss of
nuclear ATRX expression in the tumor cells. Two cases showed mosaic loss of
H3K27me3 expression in neoplastic cells. The Ki-67 proliferation index was
significantly high (40-80%). Variable neuronal markers, such as Syn, NF,
SOX2, MAP2, were expressed in all tumor samples. Genetically, all tumors
samples harbored two DICER1 alterations, including a hotspot missense
mutation in the RNase IIIb domain within exon 25 on
one allele (p.E1813 or p.D1810) and a copy number deletion or mutation on the
other allele (a germline mutation in one case). In addition, sarcomas showed
alterations in TP53 (80%), ATRX (60%), and the mitogen-activated protein
kinase pathway genes (60%). Finally, all five cases were diagnosed as
DICER1-mutant primary intracranial sarcoma. All patients underwent craniotomy
resulting in complete tumor resection. Patient 2 succumbed to the tumor after
3 months post-surgery due to rapid progression and tumor dissemination. Three
patients accepted radiotherapy and chemotherapy following surgery, with progression-free
survival times of 28, 48, and 50 months, respectively. One patient was lost
to follow-up 3 months after surgery. Conclusion DICER1-mutant primary
intracranial sarcoma, a newly defined tumor entity in the fifth edition of
the WHO classification of central nervous system tumors, commonly occurs in
children and young adults. High-grade malignant spindle cell sarcoma and
eosinophilic cytoplasmic globules are their typical morphological features.
Besides myogenic differentiation, these tumors may also exhibit neuronal
differentiation. Confirming the pathological diagnosis relies on genetic
testing for DICER1 pathogenic alterations or DNA methylation profiling. Key words: DICER1; Intracranial;
Sarcoma; DNA methylation; Eosinophilic globule |
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El Helali,
Aya The University of Hong Kong Combination Talazoparib
- carboplatin for recurrent high-grade glioma with DNA damage repair
deficiency (DDRd) “TAC-GReD”
trial Background: Recurrent high grade gliomas (HGG)
present a significant challenge in neuro-oncology due to their poor prognosis
and lack of effective standard treatment. Emerging evidence indicates
deficiencies in the DNA damage repair (DDR) pathway in a significant
proportion of gliomas, leading to innate or acquired resistance mechanisms
and worse survival outcomes. Somatic alterations in the DDR pathway,
including PTEN mutations and ""BRCA-ness"", have paved
the way for targeted therapy utilizing poly (ADP-ribose) polymerase (PARP)
inhibitors. The distinct mechanisms of action of PARP-trapping inhibitors,
particularly in forming cytotoxic PARP-DNA complexes, have shown promise in
personalized cancer treatment strategies. Methods: The TAC GReD
trial is a prospective phase II, single-arm, open-label study designed to
evaluate the efficacy and safety of the combination of talazoparib
and carboplatin in patients with recurrent HGG harboring pathogenic mutations
in the DDR pathway. The trial aimed to assess the 6-month progression-free
survival (PFS-6) as the primary endpoint, with secondary endpoints including
objective response rate (ORR), overall survival (OS), safety and
tolerability, and health-related quality of life (HRQL) measures. Results: Thirty-three patients, with a
median age of 55 and 70% male, were enrolled in the TAC GReD
trial. The trial met the prespecified response criteria, with a PFS-6 survival
rate of 25% (95% CI 13–48%), median progression-free survival of 3.4 months
(95% CI 2.2–5.7%), and overall survival of 6.5 months (95% CI 4.5–9.8). The
treatment combination was deemed safe, with the most common TRAEs being
hematologic, and no treatment-related deaths were reported. The quality of
life scores remained stable throughout the trial. Conclusion: The unique biomarker-stratified
approach capitalizes on the strong biological rationale for utilizing PARP
inhibitors in HGG, offering a potential avenue for innovative therapeutic
strategies in the management of recurrent HGG. |
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Fang, Deyu Northwestern University A novel therapy with onco-targeting
and immune-boosting dual efficacies in combating a broad spectrum of human
cancers Immunotherapies have revolutionized
the treatment landscape for various solid tumors. However, only a subset of
cancer patients benefit from the current checkpoint blockade therapies.
Recent research underscores the impact of an immune-suppressive tumor microenvironment,
marked by increased Treg infiltration and limited neoantigen presentation, as
key factors driving resistance to immune checkpoint blockade. Furthermore,
the integration of immune checkpoint inhibitors with standard chemotherapy,
termed chemoimmunotherapy, has emerged as a cornerstone approach for several
cancers. However, the immune-depleting toxicity of chemotherapeutic regimens
often undermines chemoimmunotherapy efficacy. This highlights an urgent need
for innovative precision therapies that effectively target both the tumor and
bolster the immune response to overcome this resistance. Our laboratory has
pioneered research on ubiquitin-specific peptidase 22 (USP22) over the past
decade, establishing it as an oncogene (Molecular Cell, 2012). More recently,
we demonstrated that USP22 promotes Treg-mediated immune suppression (Nature,
2020), identifying it as a dual-action target to both inhibit tumor growth
and enhance antitumor immunity. Building on these insights, we developed the
first USP22-specific small molecule inhibitor, which demonstrates both
oncogenic targeting and immune-boosting activity with minimal toxicity across
multiple solid tumor types (Science Advances, 2022). Our latest research
further shows that USP22 enables tumors to evade immune surveillance by
suppressing neoantigen presentation, providing a mechanistic basis for
targeting USP22 to overcome immunotherapy resistance. With these insights, we
have advanced to the development of a second-generation USP22-specific
inhibitor with markedly improved antitumor efficacy. We are currently
completing IND-enabling studies, with the ultimate goal of conducting the
first-in-human Phase 1 clinical trial of this USP22-specific inhibitor in
solid tumors, including glioblastoma. |
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Fong, Vernon Hospital for Sick Children Defining the involvement of the
perivascular niche in brain tumor metastases Medulloblastomas (MB) are the most
prevalent malignant brain tumors in children, arising in the cerebellum or
dorsal brainstem. These tumors are categorized into four distinct subgroups:
WNT, SHH, Group 3, and Group 4, each characterized by unique gene expression
profiles, metastatic behaviors, and recurrence rates. Importantly, metastatic
progression is often associated with poor prognosis, impacting about 30% of
patients at diagnosis. Despite extensive research on primary tumors, there
are currently no approved treatments specifically targeting MB leptomeningeal
metastases. Consequently, enhancing our understanding of metastatic MB is
essential for developing targeted therapies. The leptomeningeal niche, where
metastatic MB cells reside, is comprised of a complex network of blood
vessels supported by perivascular cells, including pericytes, smooth muscle
cells, and fibroblasts. However, the specific adhesion molecules and
signaling pathways that facilitate tumor cell colonization in this niche
remain largely undefined. In this study, we hypothesize that the perivascular
niche plays a critical role in the colonization and survival of metastatic cells
within the leptomeninges. To investigate this hypothesis, we
utilized an innovative sLP-mCherry niche labeling
system alongside single-cell RNA sequencing to spatially identify and analyze
the perivascular cells surrounding metastatic MB tumor cells. Our analysis
revealed upregulated genes linked to cholesterol export in these
tumor-associated perivascular cells, indicating their role in facilitating
metastatic cell colonization. To further explore this mechanism, we employed
CRISPR and lentiviral shRNA constructs to knock down lipid receptors in MB
cells, thereby reducing cholesterol uptake from the leptomeningeal
microenvironment, which led to significant decrease in metastatic burden.
Additionally, we observed that inhibiting perivascular recruitment to the
tumor site—by decreasing chemokine secretion in MB cells—resulted in a
reduced metastatic burden. In summary, our study underscores
the importance of the perivascular niche in metastatic medulloblastomas and
provides novel insights into the molecular mechanisms driving tumor cell
colonization within the leptomeninges. By exploring lipid transport
mechanisms in the vascular microenvironment, we open new avenues for
developing innovative treatments targeting MB metastases. |
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Giamas, Georgios University of Sussex & Zhejiang
Chinese Medical University Developing an extracellular
vesicles-based liquid biopsy biomarker signature of glioblastoma and
comparison to whole genome sequencing plus tissue proteomics data using
samples from the Genomics England Biobank Background: Despite significant advancements in
the understanding of gliomagenesis, there remains
no glioblastoma subtyping framework that is used in clinical practice.
Additionally, diagnosis requires an invasive tissue biopsy, which limits the
ability to perform longitudinal sampling resulting in a more limited
understanding of the changes that occur in response to treatment and over
time compared to other solid tumours. Extracellular vesicles
(EVs), small lipid membrane bound particles that contain various bioactive
molecules including proteins, RNA and DNA, have been proposed as a potential
liquid biopsy biomarker as their cargo has been demonstrated to reflect their
cell of origin. Through the integration of plasma derived-EVs proteomics,
transcriptomics and Raman spectroscopy signature, whole genome sequencing
data, and tissue proteomics we aim to identify biologically and clinically
relevant glioblastoma subtypes. Methods: Using 1mL glioblastoma plasma
samples from the Genomics England biobank, and sex- and age-matched volunteer
plasma samples, EVs were separated by size exclusion chromatography and characterised by protein quantification, nanoparticle
tracking analysis, transmission electron microscopy and Western blot analysis
to confirm EV enrichment. Differences in yield and particle size were
assessed using an unpaired Welch’s t-test. Subsequently, analysis of the
proteomic and transcriptomic cargo and label free Raman spectroscopy of the
isolated EVs was investigated and bioinformatic analysis performed to
identify a glioblastoma biomarker signature. Further comparisons with the
whole genome sequencing data and tissue proteomics were investigated. Results: Plasma samples from glioblastoma
patients from the Genomics England biobank have been analysed.
Patients were predominantly male (63%) with a median age of 53. EVs have been
separated and characterised from 57 glioblastoma
and 49 healthy volunteer plasma samples. Following EV separation there was no
difference in the mean protein yield (362.4 vs 244.4 μg
protein, p=0.5860) or mean particle yield (1.28 x1011 vs 1.18 x1011
particles, p=0.5645) from 1mL plasma samples between glioblastoma patients
and healthy volunteers. However, there was a significant difference in the
mean (133.6 vs 114.9 nm, p<0.0001) and median (121.5 vs 103.4 nm,
p<0.0001) particle size which is suggestive of altered EV biogenesis. Multi-omic
analysis of ~1000 identified EV protein species and EV micro-RNA cargo
identified significant differences between glioblastoma and healthy volunteer
plasma EV samples. Bioinformatic analysis, using a machine-learning
algorithm, incorporated the identified multi-omic
EV cargo plus Raman spectroscopic signature into a glioblastoma biomarker
signature. The developed EV biomarker
signature will be added to clinicopathological, treatment and whole genome
sequencing data to develop a comprehensive prognostic model. The identified
signature will also be compared to tumour
tissue-based mass-spectrometry proteomics and tissue spatial proteomics to
investigate novel glioblastoma subtypes and the cellular source of the
components of the identified biomarker signature. Conclusion: The development of an EV liquid
biopsy signature for glioblastoma will have numerous applications, including
a role in early diagnosis, development of a prognostic/predictive biomarker
model, and patient monitoring during and following treatment. Additionally,
novel biological insights related to intrinsic glioblastoma subtypes and tumour biology are likely to be identified. |
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Hassuneh, Osama the University of Hong Kong Developing Lipid Nanoparticles for
Combinatory Chemo-Immunotherapy of Glioblastoma Glioblastoma (GB) is an aggressive
type of glioma, treated by a combination of surgery, radiotherapy and
chemotherapy. However, its prognosis is still dismal. In recent decades,
immunotherapy has revolutionised cancer treatment.
Nonetheless, as monotherapy, immune modulation did not significantly improve
treatment outcomes, as multiple clinical trials demonstrated disappointing
results, specifically for GB patients. The GB tumour
microenvironment (TME) is enriched with myeloid-derived suppressor cells, in
addition to GB cells expressing surface proteins, such as CD47, which inhibit
macrophage-mediated phagocytosis. Immunogenic cell death (ICD) is a specific
apoptotic mechanism in which dying cells release stress molecules that
trigger immune cell maturation and activate the adaptive immune response. This research focuses on developing
lipid nanoparticles for co-delivery of small interfering RNA (siCD47) to
downregulate CD47, alongside an ICD inducer to boost the immune response within
the microenvironment of GB. Lipid nanoparticles (LNPs) have
recently attracted attention as efficient carriers of nucleic acids (NA) for
a wide range of applications, from COVID-19 vaccines to gene therapy agents
providing a high encapsulation efficiency (EE) with minimal cytotoxicity.
Here, LNPs co-encapsulating siCD47 and an ICD-inducer drug, including
doxorubicin (DOX), mitoxantrone (MIT) or oxaliplatin (OXA), were prepared
with a biocompatible size of (< 140nm) polydispersity index of (< 0.2)
and a high %EE of siCD47 (> 90%). Drug encapsulation into LNPs was optimised via statistical analysis and mathematical
models using the Design of Experiments (DOE) approach. In vitro cytotoxicity
and ICD induction ability of the optimised LNPs
were evaluated and compared with free drugs against brain cancer stem cells
(BL6) and murine glioma cell line (GL261). BL6 cells were more sensitive to
drug toxicity compared to GL261 cells. Drug toxicity was in the order of
DOX>MIT>OXA. DOX and MIT encapsulation into LNPs showed reduced
toxicity due to delayed drug release. Interestingly, DOX and OXA showed
higher calreticulin (CALR) expression, an ICD marker, in the LNP form
compared to free drugs. LNPsiCD47 reduced CD47 expression in BL6 (>70%) at
doses > 10 nM. GL261 were more resilient to
silencing in vitro. LNPs encapsulating DOX and siCD47
were tested in an orthotopic GL261 mouse model following two intra-tumoral
injections of 495 µg/Kg and 2300 µg/Kg for siCD47 and DOX, respectively.
Groups receiving LNPsiCD47 and LNPsiCD47/Dox showed improved survival from 17
(untreated) to 20 and 19 days, respectively (sham Vs LNPsiCD47 or
LNPsiCD47/Dox P < 0.05, LNPsiCD47 Vs LNPsiCD47/Dox P > 0.05). In conclusion, we successfully
developed LNP formulations with therapeutic potential in the GB mouse model.
CD47 silencing shows promise in GB therapy. Although the combination with DOX
showed no added benefit in this study, future work will explore the potential
of OXA-LNP as a standalone or combination therapy. |
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Hung, Yuet Yi The University of Hong Kong Role of ENO1 in Glioblastoma
Progression Objective: To investigate whether and how ENO1
contributes to glioblastoma (GBM) progression. Method: In vitro studies were conducted
using human GBM cell lines, U87 and U251. Knockdown of ENO1 and inhibition of
ENO1 using an ENO1 inhibitor, ENOBlock, were done
on the cell lines to examine the role of ENO1 in GBM. The effects of ENO1
knockdown and inhibition were evaluated by western blot, immunofluorescence
staining, cell viability and proliferation assay, cell cycle analysis by flow
cytometry, and colony formation. Result: Treatment with ENOBlock
or knockdown of ENO1 decreases the cell viability and cell proliferation of
GBM, but increases its cell death and sensitivity to temozolomide (TMZ).
Treatment with ENOBlock also decreases
mitochondrial membrane stability in GBM. Knockdown of ENO1 reduces colony
formation of GBM and decreases the expression of mitochondrial proteins. Conclusion: ENO1 contributes to the progression
of GBM and can potentially be a therapeutic target. Coadministration of TMZ
and agents inhibiting the ENO1 signalling pathway
can potentially produce synergistic therapeutic effects. |
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Jiang, Yuanhao University of Newcastle Integrative Multi-Omics and
Histopathological Imaging for Enhanced Subtyping and Clinical Management of
Diffuse Midline Gliomas Diffuse midline glioma (DMG) is an
incurable and highly aggressive brain tumour with a
poor prognosis and limited treatment options. Currently, early diagnosis DMG
relies on the tumour’s location, histology-based
morphology, and the staining for the hallmark loss of H3K27me3 which is
inadequate for assigning molecularly distinct subtypes indictive of treatment
recommendation. High-throughput profiling of DMG specimens, has identified
genomic subtypes and potentially targetable recurring molecular features. However,
the extended time required for these multi-omics assays to be performed
limits immediate clinical application of potentially beneficial therapies,
and hence interventions used later in the clinical journey of a patient have
not yielded significant improvements in outcomes. Here, we present a comprehensive
overview of the molecular subgroups of DMG identified through advanced
multi-omics approaches – including genomics, epigenetics – as well as imaging
modalities based on histological sections. Systematically, published studies
employing molecular profiling coupled with histopathological imaging were analysed employing a novel pipeline to integrate several
distinct morphological feature analysis software’s we molecular signatures
identifying tumour heterogeneity and patient
outcomes. Our findings indicate that
integrated and matched multi-omics data and imaging offers a powerful
strategy for identifying novel subgroups and actionable drivers, as has been
demonstrated in the case of Medulloblastoma. Furthermore, the application of
deep learning in imaging data analysis shows great promise in identifying
specific features, enabling the diagnosis of molecular subgroups, such as IDH
mutations in high-grade gliomas. This positions histopathological imaging as
a rapid and informative alternative to traditional molecular diagnostics. Our work emphasises
the potential of combining multi-omics data with imaging biomarkers to
enhance brain tumours subtyping, improve diagnostic
accuracy and enable personalised treatment plans.
The development of fine-tuned and robust deep learning models applied to
matched multi-omics and histopathological imaging could revolutionise
DMG management by facilitating real-time, precise subtyping and targeted
therapeutic interventions tailored to each tumour
entity. This integrated approach holds the promise of transforming clinical
outcomes for patients affected by this devastating disease. |
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Khera, Sudeep All India Institute of Medical
Sciences, Jodhpur Expression of SSTR-2 and HER-2-neu
in Meningiomas and its correlation with clinico-pathological
parameters Meningiomas are one of the most
common tumours of the central nervous system.
Although commonly benign, atypical and anaplastic meningiomas also constitute
part of the spectrum. Options for management are limited to surgery and
radiotherapy. SSTR-2 has shown great promise as a target for therapy as well
as intra-operative imaging of meningiomas. The expression of HER2/neu status
has been substantial in various other malignancies for treatment purpose.
However, it has shown variable results in meningiomas. The present study was aimed to
evaluate the expression of SSTR-2 and HER2/neu in meningiomas and to
establish their correlation with various clinico-pathological
parameters. This was an ambispective
observational cross-sectional study carried out in a tertiary care hospital
in Western Rajasthan. This study included 120 cases of meningioma. On
application of immunohistochemistry for SSTR-2 and HER2/neu on these cases,
75.83% cases showed positivity for SSTR-2 and none of the cases were positive
for HER2/neu. SSTR-2 expression showed significant association with location
of meningioma in brain and with histological subtypes. No significant
association was found with age, gender, specific histo-morphological
features and tumour grade. None of the cases in this study
were positive for HER2/neu. Previous studies also have showed a highly
variable expression of HER2/neu in meningiomas. Hence, more studies are
needed to investigate the role of HER2/neu in meningiomas as a diagnostic and
prognostic marker as well as its role in targeted therapy. Expression of SSTR-2 has been
recommended by the 2021 WHO classification of CNS tumours
as a desirable criteria for diagnosis of meningioma. In the present
study,75.83% cases were positive for SSTR-2, however, a significant
association with the grade and other clinical parameters could not be
established. Since survival analysis was not performed, correlation with prognosis
could not be assessed. Therefore, further studies are recommended to
establish a significant clinico-pathological
correlation as somatostatin receptor may have a significant role in
confirmation of diagnosis of meningioma on imaging and in targeted therapy-based
management in such cases. |
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Killilea, Joshua Imperial College From Innovation to Treatment:
Bridging the Gap Between Engineering and Clinical Practice in Brain Tumor
Therapy Brain tumours
are a group of malignancies characterised by high
morbidity and mortality. The most common and aggressive primary brain tumour in adults is glioblastoma multiforme (GBM). For
the past two decades, survival outcomes for GBM have remained stagnant with
traditional avenues of surgery, chemotherapy, and radiotherapy being largely
ineffective with the median survival time being 15 months. There is therefore
an urgent need to develop improved treatment options for GBM patients.
Developing novel pharmacological agents for GBM, however, represents a
challenging and unrewarding prospect. As such, only 0.1% of novel anticancer
agents for GBM progress from Phase 1 trials to registration. For other
cancers, this number is 9%. Furthermore, developing novel pharmacological
agents is hugely resource intensive from a time and financial perspective.
Attempts to improve treatment outcomes in GBM, as well as other brain tumours, will therefore benefit from input from engineers
to develop strategies to overcome current treatment limitation in GBM. For
example, it is possible to develop stimuli-responsive vehicles for local drug
delivery using ‘smart’ biomaterials, focussed
ultrasound to transiently open the BBB and tumour
treating fields. While such promising treatment options are currently being
explored for GBM, critical to their successful translation will be effective
interdisciplinary collaborations between engineers and clinicians. Such
interdisciplinary collaboration is not always effective and can be
challenging due to a significant gap in the mutual understanding of
terminology as well as different ideas on which research questions are
clinically important to address. We are conducting a global survey directed
at clinicians and engineers to elucidate crucial insights regarding the
quality of communication, comprehension of technical requirements and
constraints, and the alignment of clinical needs with engineering solutions.
Overall, the results demonstrate that clinicians are largely supportive and
aware of the need for engineering advances to address challenging questions
in cancer research, with all respondents agreeing with the statement that
engineering input will be critical in addressing questions in cancer
research. Furthermore, useful strategies for more effective inter-disciplinary
collaborations are presented. Common strategies suggested included the need
for a ‘common language’ between clinicians and engineers as well as shared
reading material to help create shared research goals. Another key point was
that clinicians need to continue to network with engineers and provide
succinct summaries for them on current clinical challenges to set the scene
for where engineers can help. To help facilitate the translation of such
developed technologies, clinicians also think it would be useful to attend
their clinical practice to see better the clinical workflow to ensure that
engineers design something that is not also effective but clinically
feasible. This survey highlights the imperative for a standardized framework
to enhance engineer-clinician collaboration, thereby maximising
the clinical impact of technological advancements in the treatment of brain tumours. |
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Lan, Yanjie Beijing Tiantan
Hospital, Capital Medical University SYHA1813, a dual-target inhibitor
of VEGFR and CSF1R, exerts antitumor activity in meningioma in vitro and in
vivo Meningioma is common tumors of the
central nervous system, arise from the dura mater and the outer layer of the
arachnoid. Malignant meningiomas are rare subtype, but they are generally
more aggressive with high recurrence rate. Malignant meningiomas patients
still lack effective therapy currently. Therefore, developing a novel regent
is urgently needed. SYHA1813 is a novel dual-target inhibitor, which targets
CSF1R and VEGFR. Our previous study suggested SYHA1813 exhibits potent
preclinical antitumor effects in glioblastoma. It’s also reported that
SYHA1813 has potentiality in treating B-cell lymphoma. However, the actual
functional role of SYHA1813 in meningiomas remains unknown. Herein, we
investigated the tumor inhibition effect of SYHA1813 on meningioma cells in
vitro and in vivo. To explore the cell toxicity of
SYHA1813 in meningioma cells in vitro, we evaluated the anti-proliferation
effect of SYAH1813 on two human meningioma cell lines. CCK8 assay results
showed that SYHA1813 reduced the cell viability of IOMM-Lee and CH157 cells
in a dose-dependent manner. Consistent with CCK8 assay, EdU
assays showed SYHA1813 treatment resulted in a marked decrease in the
fraction of EdU-positive cells. In addition,
SYHA1813 suppressed colony formation of IOMM-Lee and CH157 cells, indicating
that SYHA1813 could affect the long-term survival of meningioma cells. Wound
healing assay and trans-well cell invasion assay revealed that SYHA1813
remarkably inhibited the migration and invasion of IOMM-Lee and CH157 cells.
These results demonstrated SYHA1813 exhibited significant anti-proliferation
activity in meningioma cells. To determine whether the inhibitory
effects of SYHA1813 on proliferation were due to cell cycle arrest or
apoptosis. Meningioma cells were treated with different dose of SYHA1813 for
48h and analyzed the cell cycle and apoptosis by flow cytometry. We found
SYHA1813 arrested cell cycle at G2/M phase and promoted cell apoptosis. Interestingly,
enlarged and flattened IOMM-lee and CH157 cells following exposure to
SYHA1813, which appealed us to explore whether SYAH1813 could induce cell
senescence. SA-β-Gal staining assays showed that the augmentation of
senescent cells, indicating that SYHA1813 could promote cell senescence in
meningioma cells. Taken together, these findings suggested that SYHA183
treatment could block cell cycle, promote cell apoptosis and senescence,
which leads to an inhibition of the proliferation in meningioma cells in
vitro. To further evaluate the functional
role of SYHA1813 in meningioma in vivo, IOMM-Lee cells were inoculated into balb/c nude mice to monitor subcutaneous xenograft mouse
model. We found tumor volume and weight of mice orally administered with SYHA1813
was effectively reduced compared to mice in vehicle group. Tumor cell
proliferation was assessed by Ki67 immunohistochemistry (IHC) staining. Less
Ki67-positive cells were observed in tumors from mice treated with SYHA1813.
Collectively, consistent with in vitro studies, these results explained the
role of SYHA1813 in inhibiting meningioma cell growth in vivo. In summary, our study revealed the
antitumorigenic role of SYHA1813 in meningioma cells in vitro and in vivo,
proving the great potential of SYHA1813 in the treatment of malignant
meningioma. |
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Lv, Jing Beijing Normal University Loss of temporally regulated
differentiation of neural stem cells in IDH-wildtype glioblastoma Background: Isocitrate dehydrogenase
(IDH)-wildtype glioblastoma (GBM) is highly heterogeneous and aggressive
brain tumor in adults. Though understanding of molecular pathogenesis of
IDH-wildtype GBM has been advanced, patients´ survival outcomes have not been
significantly improved. Mapping GBM into neural lineages and differentiation
stages may uncover hitherto inadequately characterized therapeutic targets.
Based on the expression profile of the EGFR or PDGFRA co-expression module
(the EM and PM, respectively), we proposed an EM/PM classification scheme and
assigned adult gliomas into EM or PM molecular subtypes that are distinct in
clinical phenotypes, genomic alterations, and resemblance to neural lineages.
EM subtype gliomas correspond to IDH-wildtype GBM; PM subtype gliomas contain
IDH-mutant astrocytoma and oligodendroglioma. EM and PM subtypes are stable
during progression of individual patients and across the regions and single
cells of individual gliomas. Glioma progression is not driven by a switch of
molecular subtypes but by genomic instability and microenvironment. Here we
mapped the aberrant neural developmental program of IDH-wildtype GBM. Methods: Using bulk transcriptomes,
single-cell transcriptomes and proteomes, GBMs were mapped into neural
lineages. Expression patterns of signatures of neural stem cells (NSC) and
neural progenitor cells (NPC) in individual IDH-wildtype GBM cells were
assessed using single-cell RNAseq analysis and RNAscope on clinical samples. We assessed the expression
patterns of key regulators in neuronal and oligodendrocyte lineage in GBM
transcriptomes. Conversely, we also assessed the expression pattern of
IDH-wildtype GBM-enriched genes in the transcriptomes of brain development. Results: Signatures of NSC and neural NPC
are enriched in the transcriptomes and proteomes of IDH-wildtype GBMs. In
contrast, signatures of neuronal and oligodendrocyte lineage are
significantly down-regulated in IDH-wildtype GBMs. IDH-wildtype GBM cells are
correlated to NSCs and NPCs. Canonical markers of NSC and NPC are enriched in
distinct cell populations of IDH-wildtype GBM cells. Regulators of fate
decision and development of neuronal and oligodendrocyte lineages are
downregulated in IDH-wildtype GBMs. Expression patterns of IDH-wildtype
GBM-enriched genes during human brain development indicate loss of temporally
regulated differentiation of neural stem cells. Expression pattern of those
temporally dysregulated genes in mouse brain development indicates precocious
astrocyte genesis in IDH-wildtype gliomas, which coincides with abnormal
activation of Notch and STAT signaling. Conclusions: Our studies show that IDH-wildtype
GBMs resemble NSC and NPC and are blocked from differentiation into mature
neuron or oligodendrocyte. Abnormal activation of Notch and STAT signaling in
NSC may induce precocious astrocyte genesis and inhibit neuronal and
oligodendrocyte differentiation, eventually cause IDH-wildtype GBM genesis.
These findings indicate that restoring differentiation could be an
alternative therapeutic approach for IDH-wildtype GBM. Key words: IDH-wildtype GBM; EM/PM
classification; brain development; neural differentiation |
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Maas, Sybren Leiden University Medical Center IDH-mutant astrocytomas with primitive
neuronal component have a distinct methylation profile and a higher risk of
leptomeningeal spread IDH-mutant astrocytomas are diffuse
brain tumours that are defined by characteristic
mutations in IDH1 or IDH2 and do not have a 1p/19q co-deletion. The
established grading criteria include histological features of brisk mitotic
activity (grade 3), necrosis or microvascular proliferation (grade 4). In
addition, a homozygous deletion of the CDKN2A/B locus is implemented as a
molecular marker for grade 4 IDH-mutant astrocytomas.
We describe a subgroup of IDH-mutant astrocytomas defined
by primitive neuronal morphology based on histology and a distinct DNA
methylation profile (n=51, ASTRO PNC). Misinterpretation as carcinoma
metastasis was common, since GFAP expression was absent in the primitive
neuronal component, whereas TTF1 expression was detected in n=14/18 (78%) of
cases based on immunohistochemistry. Targeted DNA panel sequencing revealed
enrichment for alterations in RB1 (n= 19/51, 37%) and MYCN (n= 11/51, 22%)
besides the astrocytoma-characteristic mutations in IDH1, TP53 and ATRX.
Homozygous CDKN2A/B deletions (n= 1/51, 2%) and CDK4 amplifications (n= 3/51,
6%) were relatively rare events in our cohort. The clinical (n=31) and survival (n=23) data we collected
indicate a biological behaviour similar to WHO
grade 4 IDH-mutant astrocytomas (p= 0,2446) with an
increased risk for leptomeningeal (n=7) and extra-CNS (n=2) spread. Taken
together, ASTRO PNC is defined by a distinct molecular and histological
appearance that can mimic metastatic disease, featuring an aggressive clinical
course. |
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Mahdi, Haridini
Intan Setiawati Indonesian National Cancer Center-Dharmais Cancer Hospital, Jakarta, Indonesia Clinical Characteristics of
Pediatric Brain Tumors : A Single Institution Experience in NCC Dharmais Cancer Hospital, Indonesia Background: Pediatric brain tumors are the most
common solid tumors among children aged 0–14 years and remain a leading cause
of childhood cancer mortality. The clinical characteristics of these tumors
are diverse, encompassing variations in developmental origin, genomic
profiles, therapeutic approaches, and outcomes. While advancements in
surgical techniques, neuro-oncology, neuroradiology, and radiation oncology
have significantly improved survival rates, certain pediatric brain tumors
continue to have poor prognoses. Understanding these clinical characteristics
is essential for enhancing diagnosis, management, and prognosis of pediatric
brain tumors. Objective: To describe the clinical
characteristics of pediatric brain tumor patients treated at the National
Cancer Center, Dharmais Cancer Hospital, Jakarta,
Indonesia. Methods: This cross-sectional descriptive
study analyzed medical records from Dharmais Cancer
Hospital between July 2021 and June 2024. Using consecutive sampling, 79
pediatric brain tumor cases were reviewed. Results: Among the 79 cases that met the
study criteria, most patients were male (64.5%) and were predominantly
diagnosed at ages 0-9 years (56.9%). Headache was the most common presenting
symptom, leading to hospital admission (39.2%). Tumor location was frequently
found in the cerebellum and fourth ventricle (36.7%), with gliomas (30.3%),
medulloblastomas (26.5%), and ependymomas (15.1%) identified as the most
common histological types. The majority of patients (37.9%) received a full
combination of therapies, including surgery, chemotherapy, and radiation.
Despite comprehensive treatment, outcomes were sobering, with 46.8% of
patients succumbing to the disease, 43.0% surviving, and 10.1% lost to
follow-up. The median time to diagnosis was 3 months, and the median
treatment initiation time was 1 month. Conclusion: This study highlights a high
prevalence of brain tumors among young pediatric males presenting with
headaches as a primary symptom. Tumors predominantly affected the cerebellum
and fourth ventricle, with glioma being the most frequent histological type.
While multi-modal treatments are widely utilized, the high mortality rate
underscores an urgent need for improved diagnostic and treatment strategies.
Enhanced understanding of these clinical characteristics can guide future
research and therapeutic innovations, ultimately aiming to improve survival and
quality of life for pediatric brain tumor patients in Indonesia. |
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Mai, Hoang Vu Tam Anh General Hospital Evaluation of results in
intracranial gliomas surgery by using synaptive
Modus V robotic system at Tam Anh General Hospital Ho Chi Minh City in
Vietnam Background: Glioma is one of the common primary
intra-axial tumors in CNS. The age-standardized incidence rate of gliomas was
4.7 per 100,000 person-years. Glioma tumor surgery has two goals: removed
tumor and preserved neurological functions. The Modus V Synaptive
robot includes an exoscope system with the robotic
arm that integrates a modern navigation system in neurosurgery. This
navigation system can regenerate the nerve fiber bundles (DTI) during
operation, allowing the neurosurgeons to access the lesions without affecting
the nerve conduction bundles for surgery. Objective: To evaluate the results of gliomas
surgery using the Modus V Synaptive robot system at
Tam Anh General Hospital HCMC. Methods: A prospective descriptive study of
a series of cases with a diagnosis of gliomas operated by the Modus V Synaptive robot system at Tam Anh General Hospital HCMC
from January 2023 to July 2024. Results: Among 43 cases were operated by
Modus V Synaptive Robot system at Tam Anh General
Hospital HCMC from 01/2023 - 06/2024. Median age’s 43.6 years. The ratio of
male: female is 1:1. Common symtomes are headache
(93.0%), followed by nausea (81.4%), weakness (65.1%), seizure (23.3%), reduce
vision (18.6%), ataxia (16.3%). Tumor locations varied, in supratentorial
with frontal (23.3%), temporal (13.9%), ventricular and thalamic involvement
9.3% of cases, followed by basal ganglia and caudate nuclei (7.0%), parietal
(7.0%), occipital (7.0%), optic pathway (7.0%), hippocampus (4.7%), and
pineal region (4.7%) in- volvement. For
infratentorial, the tumors were located in cerebellar hemisphere, vermis and
brainstem (9.3%, 4.7% and 2.3% respectively). On the preoperative MRI- DTI,
the change of tracts with tumors were divided into 4 types: normal (14%),
displacement or edematous (48.8%), infiltration (27.9%), discruption
or destroyed (9.3%). Transsulcal parafascicular using tubular retractors (Vycor, NICO) was performed in 21 cases (48.8%). The
histopathology of tumors were classified with grade I involvement 15 cases
(34.9%), followed be grade II (32.6%), grade III (14.0%) and grade IV
(18.5%). Gross total resection (GTR) was achieved in (60.5%) of cases,
near-total resection (NTR) in (23.2%), and subtotal resection/partial
resection (STR/PR) in (16.3%). The recovery of neurological functions
significantly different between GTR and STR/NTR/PR (p<0.0001).
Postoperative complications included hemiparesis or weakness (11.6%), CFS
leak (9.3%), infections (7.0%), visual deficits (4.7%), seizure (4.7%). The
GOS score at discharge from the hospital was at 5 :
34 cases (79%), followed by GOS3-4 with 6 cases (14.0%), GOS1-2 with 3 cases
(7.0%). Conclusion: The neurotransmission recovery
after surgery is related to the patient's discharge GOS score. The GTR has
been showed good results and clinical patient improved. Robotic surgery
achieves the positive effect in brain tumor surgery when nerve fiber bundles
are preserved as much as possible. |
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Miceli, Giovanni Carlo Imperial College London Overcoming the Blood-Brain Barrier:
A Polymeric Bioelectronic Implant for Targeted, Voltage-Driven Drug Delivery Brain tumours
are characterised by high morbidity and mortality
owing to their localisation and often locally
invasive growth. Glioblastoma multiforme (GBM) represents the most common
malignant tumour in the central nervous system
(CNS), accounting for 14.2% of all tumours and
50.9% of all malignant brain tumours. Despite
advances in neurosurgery, chemotherapy, and radiotherapy, it remains one of
the most treatment-resistant CNS malignancies, and the tumour
inevitably recurs. Survival rates for GBM have remained unchanged since the
introduction of the Stupp protocol almost 20 years ago. The prognosis is only
15 months, with less than 5% of patients surviving beyond 5 years even with
aggressive treatment. Attempts to design pharmacological agents capable of
treating glioblastoma are hampered by tumour
heterogeneity, immune privilege of the central nervous system and the
blood-brain barrier. The primary chemotherapeutic agent used to treat these tumours is temozolomide due to its ability to cross the
blood-brain barrier (BBB). The BBB is a pivotal obstacle to GBM treatment
since it regulates the entry of molecules into the brain. Local drug delivery
platforms offer the ability to get drugs to target cells while minimising systemic delivery and adverse events
associated with treatment. Some of the recent devices developed for tissue localised clinical administration of drugs have been
focused on using pressure to inject drug solutions into the diseased tissue
area. This method relies on pressure differentials causing increased
intracranial pressure and drug reflux. To address this challenge, we have
developed a polymeric bioelectronic implant to control the drug concentration
in the tumour microenvironment by leveraging the
voltage application. This strategy aims to overcome chemoresistance and unnecessary
exposure to healthy tissues by circumventing the BBB for high-precision
chemotherapeutic targeted drug delivery. This bioelectronic implant consists
of a polyurethane matrix with a conductive polymeric filler for
voltage-controlled dry release of drug molecules. The material enables the
encapsulation and stabilising of small molecule
drugs and on-demand electrically controlled offloading of the drugs.
Proof-of-concept has been demonstrated using anionic and cationic dyes,
demonstrating that their release can be enhanced at repulsive and inhibited
at attractive voltages. This device has the potential to load several charged
chemotherapeutics and precisely release them within the tumour.
Preliminary experiments have been performed using doxorubicin, paclitaxel and
cisplatin, using voltages well within the range of safe human levels,
demonstrating that we are indeed able to control the release of anionic and
cationic chemotherapeutic drugs. Given the tumour
heterogeneity of GBM, this feature is crucial to target multiple tumourigenic molecular pathways to reduce chemoresistance
and recurrence. The implant uses electrophoretic release to control the
unloading of charged chemotherapeutic agents and can improve tissue
penetration over passive local delivery approaches. Moreover, electrical
stimulation through the drug-releasing electrode may allow modulation of
proliferative potential and drug sensitivity. Multimodal treatment through
locally targeted chemotherapy and electrical modulation of cancer cell biology
may yield unexpected synergies that can finally make a significant step
towards curing patients. |
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Nguyen, Nhu Children's Hospital 2 Clinical Characteristics and
Treatment Outcomes of Intracranial Germ Cell Tumor at Children's Hospital 2
in the South of Vietnam Objective: Intracranial germ cell tumors
(IGCTs) are an uncommon neoplasm that predominantly impacts children and
young adults. Pediatric IGCTs represent a significant challenge in low-income
nations due to restricted access to specialized care and treatment resources
may be limited. This study aims to provide essential information regarding
Characteristics, Clinical Features, Treatment Approaches, and Outcomes of
Intracranial Germ Cell Tumor at Children's Hospital 2. Method: A retrospective analysis of
twenty-seven pediatric patients diagnosed IGCTs at Children’s Hospital 2 in
Ho Chi Minh City, Vietnam, from May 2020 to November 2022, with follow-up
extending until May 2023. Results: The study found a male-to-female
ratio of 4.4:1 among participants, with a mean age of diagnosis at ten years.
The median interval from symptom start to diagnosis was two months. The most
commonly reported symptoms were headache and nausea or vomiting, impacting
approximately 52% of the patients. Subsequently, localized neurological
deficits manifested in around 33% of cases, whereas endocrine disturbances
were observed in 22% of subjects. Magnetic resonance imaging indicated that
the most common locations of tumors were the pineal and pituitary glands,
accounting for 44% and 33% of cases, respectively. No cases of metastasis
were detected at the time of diagnosis. The number of patients with increased
AFP and beta HCG in blood and cerebrospinal fluid and without increased AFP
and beta HCG was almost equal, at 49% and 51%, respectively. Endocrine
disorders, especially hypopituitarism, were common in patients with pituitary
gland tumors, impacting 78% of this group. Following the guidelines and
protocols of the Société Internationale d'Oncologie Pédiatrique Central
Nervous System Germ Cell Tumor II (SIOP CNS GCT II), the majority of patients
received surgical intervention, with nine patients undergoing partial
resection (33%) and six patients undergoing total resection (23%). In
addition to surgery, all patients received chemotherapy, and twenty patients
(74%) experienced radiotherapy as part of a multimodal treatment approach.
The 12-month survival rate was 76%, with a median survival duration of 23
months. Notably, no tumor recurrences were observed by the end of the study
period. Conclusion: This study offers significant
insights into the clinical presentation, treatment, and outcomes of pediatric
IGCTs at Children's Hospital 2, Vietnam. Despite constrained resources in a
low-income environment, the findings have the potential for favorable
short-term outcomes using a multimodal treatment strategy comprising surgery,
chemotherapy, and radiotherapy. |
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Pitchaimani, Arunkumar Vellore Institute of Technology
(VIT) Mitochondrial transplantation: an organelle-mediated
drug delivery system for potential glioblastoma multiforme therapy Organelle-mediated drug delivery is
a novel bioengineering approach exploiting the usage of the mammalian
cellular organelle to deliver therapeutic drugs/genes for various biomedical
applications. Among them, mitochondria(MT), a
powerhouse of the cell play a crucial role in activating the apoptosis of the
mammalian cells. With the advent of MT transplantation technology, MT has
been exploited to overcome the current clinical challenges in many
mitochondrial diseases. However, in cancer therapy, most of the
drugs/therapeutics are more focused on only targeting MT in cancer cells.
Based on the clinical layouts and the biopotential of MT-transplantation, we
hypothesized that MT-transplantation in the presence of the anticancer drug
will enhance apoptotic signatures in tumor cells. The utilization of MT as a
drug delivery vehicle has not been explored so far. In this study, we
investigated the drug-loading strategies of functional MT and studied its
anti-cancer efficacy against glioblastoma multiform model in mice models. For
this, a functional MT from Mesenchymal stromal cells was isolated and
characterized for its stability through size, morphology, membrane potential,
ATP production, etc. Further, MT was loaded with the model chemotherapeutic
drug, doxorubicin. Using bioengineering approaches, we developed novel
strategies to load higher concentrations of drug molecules into mitochondria
without affecting their viability and membrane integrity. The novel
strategies include (1) 37⸰C incubation; (2) Saponin treatment (3) Freeze-Thaw (4)
Electroporation. With DOX-loaded-MT, we investigated its
in vitro anti-tumor potential in U87MG cell lines and also evaluated its
PK-PD parameters, brain accumulation potential, and in vivo anti-glioblastoma
efficacy in preclinical mice models. The results showed that the functional
mitochondria were successfully isolated from the MSC cells and characterized
for their size, yield, and protein concentrations. With our experimental
setup, we also found the exogenous cell-free mitochondria can also be taken
up by the cell through the process of endocytosis. Further results show
enhanced cellular uptake and intracellular distribution in U87MG after 6h
incubation. It may be envisioned that supplying energy to the cancerous cells
in the form of MT transplantation can worsen the condition, but our
preliminary in vitro results showed that donor mitochondria induce
significant apoptosis in a U87MG cell. A dose-dependent cytotoxicity and
significant apoptosis induction were observed in U87MG cells after 24h
incubation. In vivo, biodistribution shows enhanced brain accumulation
through intranasal administration compared to intravenous. The in
vivo-antitumor efficacy of the DOX-loaded-MT study is ongoing, and the
results will be compiled soon. Thus, the bioengineered mitochondria with DOX
payload will play a huge role in future glioblastoma research with enhanced
bioaccumulation in the brain for advanced glioblastoma therapy and can be
used as an effective therapeutic tool for personalized GBM therapy. |
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Plate, Karl H. Goethe University Identification and functional phenotyping of lymphoid
aggregations in brain metastases Lymphoid aggregations (LA), such as tertiary lymphoid structures
(TLS), have been shown to predict overall survival and response to immune
checkpoint therapy in non-CNS cancers. Data on tumors of the CNS, however,
are sparse. Here, we applied CD20 bright-field immunohistochemistry as a
screening tool to identify LA in a retrospective cohort of 461 FFPE-brain
metastasis samples, derived from the archives of the University Cancer Center
(UCT) Frankfurt, Germany, between 2015 and 2022. Stereotaxic biopsies as well
as spinal/vertebral metastases were excluded from the study. LA-positive
tumors were further analyzed by multispectral whole slide
TSA-immunofluorescence (IF) using antibodies for von Willebrand factor, CD3,
CD20, CD163, Lamp3, pan-cytokerarins, and DAPI as
counterstain. TSL-positive tumors were further characterized by whole exome
sequencing, bulk RNA sequencing, spatial transcriptomics and customized highplex-sequential IF panels that included > 40 antibodies
for the analysis of tumor immunology and microenvironment.
|
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Puri, Sameer All India Institute of Medical Sciences
(AIIMS), New Delhi SPECC1L-NTRK2 Fusion in High-Grade
Glioma in a Young Adult: A Rare Case Study and Diagnostic Implications Background: Neurotrophic tyrosine receptor
kinase (NTRK) gene fusions are identified in less than 2% of gliomas, with a prevalence
of less than 1% across all tumors. NTRK encodes for tropomyosin receptor
kinase (TRK), which is highly expressed in neural tissue and regulates
neuronal developmental pathways through the MAPK, PI3K/AKT and PLC-γ signalling cascades. NTRK fusions, particularly
SPECC1L-NTRK2 fusions, have been identified across various pediatric and
adult tumors, including gliomas. The clinical identification of NTRK-fusion
is important due to the availability and clinical efficacy of the newly
approved TRK inhibitors in the treatment of NTRK-fusion tumors and shows good
prognosis irrespective of the tumor type, histology or grade. The detection
of these fusions via whole-genome sequencing (WGS) and whole transcriptome
RNA sequencing (RNAseq) has opened new avenues for
precision medicine, offering targeted therapies in cases where standard
treatments may be ineffective. Case History: We report the case of a 26 year old
male presenting with progressive headaches and neurological deficits. MRI
revealed a mass in the left insular region with aggressive features including
irregular thick margins, central necrosis, marked mass effect, hemorrhage,
intense irregular heterogeneous enhancement of margins with nodular
components and elevated perfusion. Observations: Surgical resection was done and
tumor tissue send for histopathological examination. Sections examined showed
features of a high grade glioma with endothelial proliferation and areas of
necrosis. On immunohistochemistry, tumor cells were immunonegative
for IDH1 R132H and p53 while ATRX was retained. MIB-1 labelling index was
10-15% in highest proliferating area. Thus, a histopathological diagnosis of
glioblastoma, WHO Grade 4 was given initially.Further
molecular profiling confirmed the presence of a SPECC1L-NTRK2 fusion,
refining the diagnosis to NTRK fused high grade glioma. Despite the
aggressive tumor characteristics, the patient showed stable disease 20 months
post-surgery, without evidence of progression. Conclusion: This case highlights the
significance of identifying SPECC1L-NTRK2 fusions in gliomas, as it opens the
door for targeted and highly effective therapies like TRK inhibitors. With
the increasing detection of NTRK fusions in various cancers, incorporating
advanced molecular techniques such as WGS and RNAseq
can significantly impact clinical outcomes, offering more personalized
treatment options. |
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Ratna, Sylvanie Dharmais NCC Hospital Serial cases : primary
neuroendocrine carcinoma (NEC) of the brain: diagnostic approach and
treatment Introduction: Primary neuroendocrine carcinoma
(NEC) of the brain is a rare case with challenging diagnostic and treatment
approach. Many cases report distinct characteristics of the patient,
treatment approach and prognosis. Our case report two different group of age with
identical immunohistochemistry result of NEC. Case Report: First case is a 70-year old patient
who visited our outpatient clinic with progressive right-sided weakness three
months before admission. He also has focal to generalized with impaired
awareness seizure. Brain MRI showed solitary brain lesion within left parietal
lobe. Patient underwent craniotomy. The tissue immunohistochemistry (IHC) was
positive for cytokeratin (AE1/3), synaptophysin, chromogranin, CD56, and
negative GFAP. CT thorax, abdomen, and pelvis showed no other lesion. Surgery
was followed by radiation therapy, 54 Gy in 30
fractions with planned brain MRI 12 weeks after radiotherapy. We preferred
not to give chemotherapy because of his age. Second case is a 21-year old woman
with increased intracranial pressure symptoms and seizure over one month ago.
Brain MRI showed solitary lesion within left frontal lobe. Tumor removal was
done and histopathology accorded with malignant tumors with papillary
pattern. IHC was positive for EMA, AE1/3, CD56, NSE, and negative for
synaptophysin, chromrogranin, GFAP, vimentin, AFP,
GATA 3, thyroglobulin, TTF-1, S100. There was no evidence of another
extracranial lesion by associating PET/CTs. A total dose of 54 Gy in 30 fractions was successfully done two months after
surgery. Chemotherapy was not performed because of poor patient compliance.
Eight months later the tumor became progressive, then tumor removal followed
by radiation therapy was repeated without chemotherapy. One year later, the
tumor progressed to double in size. Discussion: Neuroendocrine carcinoma (NEC) of
the brain is a rare case categorized as functional and non-functional. Gender
and age specific has not been reported, so do our cases that show different
patients characteristics and histopathology. Both of our patient have
identical IHC, positive AE1/3 and C56 which can be found in NEC. Tumor
removal and radiation therapy is the main treatment. Chemotherapy with
cisplatin and etoposide is recommended for high grade NEC. We suggest to
consider patient performance status, residual tumor and Ki67-positive cell,
otherwise observe until progression occurs. First patient’s Karnofsky Performance Status (KPS) is 80 and has
Ki67-positive cell >75%. Second patient’s KPS is 70 and has Ki67-positive
cell >10% but with poor treatment compliance. We decided to suspend
chemotherapy for first patient yet offer it to the second patient. Conclusion: Requirement of tumor removal and
radiation therapy of primary brain NEC is an evidence-based treatment.
Chemotherapy should be considered for high risk of recurrence cases.
Prognosis differs due to different patient characteristics. |
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Shahi, Mehdi H. Aligarh Muslim University Potential novel molecule ARSH-Q for
the treatement of most malignant brain tumor
glioblastoma Glioblastoma is still an aggressive
brain tumor and causes high mortality, despite advanced neurosurgery and
chemoradiation therapy. The relapses are common, and the main culprit is
glioma-initiating stem cells (GISCs). Targeting these stem cells is crucial
for the patient's survival and enhancement. Our lab screened multiple
compounds to better target stem cells. Fortunately, we hit a molecule ARSH-Q
which has the potential to kill glioma cells efficiently and regulate the
reduction of stem cell markers BMI1, NANOG, OCT4, SHH-GLI1 cell signaling
pathway expression and inhibited the growth of glioblastoma in vitro study.
In addition, this novel molecular/compound also has a very low LD50 compared
to the most common drug Temozolomide for glioblastoma growth inhibition. This
potential compound may be used as a therapeutic target for the treatment of
brain tumor glioblastoma. |
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Tang, Wanjun The University of Hong Kong Impaired degradation of IDH1
through Chaperone-mediated autophagy promotes glioblastoma cell cycle
progression Chaperone-mediated autophagy (CMA)
selectively targets individual proteins containing KFERQ-like motifs for
lysosomal degradation, primarily facilitated by the chaperone heat shock
cognate protein of 71 kDa (HSC70) and the receptor
lysosome-associated membrane protein type 2A (LAMP2A). While several CMA
substrates have been identified in various cancers, the specific roles and
substrates of CMA in glioblastoma (GBM) remain poorly understood. Notably,
isocitrate dehydrogenase 1 (IDH1), a key metabolic enzyme and prominent GBM
marker, has been identified as a CMA substrate in mouse embryonic stem cells.
In this study, we investigated the role of CMA in GBM and its impact on IDH1
turnover. Our results demonstrated that inhibiting CMA through LAMP2A
knockdown significantly promoted GBM cell growth both in vitro and in vivo,
while LAMP2A overexpression suppressed cell proliferation. CMA inhibition
disrupted the RB1 pathway and enhanced cell cycle progression, as indicated
by increased levels of positive regulators such as Cyclin D1 and CDK4.
Additionally, IDH1, which contains a conserved CMA motif, accumulated
following LAMP2A knockdown or lysosomal inhibition and interacted with HSC70
in GBM cells. Functionally, suppressing IDH1 reversed the tumor-promoting
effects of CMA inhibition and restored the dysregulated RB1 pathway, while
IDH1 overexpression promoted cell cycle progression. Mechanistically, we
showed that the metabolic product of IDH1, -ketoglutarate, partially stimulated Cyclin D1
expression. In conclusion, our findings suggest that CMA is essential for
suppressing GBM progression by facilitating IDH1 degradation and inhibiting
cell cycle progression. |
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Tang, Wanjun The University of Hong Kong IDH1 persistence due to compromised
chaperone-mediated autophagy accelerates cell cycle progression in
glioblastoma Chaperone-mediated autophagy (CMA)
is a selective protein degradation pathway crucial for cellular homeostasis.
However, its role in glioblastoma, a highly aggressive brain tumor, is not
well understood. In this study, we elucidate a tumor-suppressive role of CMA
in glioblastoma through the degradation of isocitrate dehydrogenase 1 (IDH1),
a pivotal metabolic enzyme and prominent molecular marker frequently
upregulated in glioblastoma. Inhibiting CMA through knockdown of its
receptor, lysosome-associated membrane protein type 2A (LAMP2A),
significantly enhances glioblastoma cell growth both in vitro and in vivo,
associated with dysregulation of the RB1 cell cycle pathway. We reveal that
IDH1 possesses a conserved CMA targeting motif and interacts with the CMA
chaperone heat shock cognate 71 kDa protein
(HSC70). Intriguingly, knockdown of LAMP2A or lysosomal inhibition leads to
IDH1 accumulation, suggesting that CMA deficiency promotes tumor growth by
scavenging IDH1 protein. Consistent with these findings, IDH1 suppression
counteracts the tumor-promoting effects of CMA inhibition and restores cell
cycle regulation in CMA-deficient cells. In contrast, upregulation of IDH1
fuels cell cycle progression, partly mediated through its metabolic product, -ketoglutarate
(-KG), which
stimulates Cyclin D1 expression. These findings reveal a previously unrecognized
role of CMA in scavenging IDH1 and regulating cell cycle progression to suppress
glioblastoma cell growth. |
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Tang, Wenshu Hong Kong Genome Institute Single-cell long-read sequencing
uncovers novel RNA isoforms and splicing variants in glioblastoma Glioblastoma is a highly aggressive
brain cancer with limited effective treatment options, driven by complex
cellular heterogeneity. Alternative splicing plays a critical role in
cellular differentiation and function, potentially driving tumor
aggressiveness and treatment responsiveness. Notable examples include
constitutive EGFR activation due to EGFRvIII
variants and exon skipping in MET, which affects sensitivity to MET
inhibitors. While short-read single-cell RNA sequencing (scRNA-seq)
is commonly used to profile cellular populations, it often lacks the capacity
to fully capture complete RNA isoforms. Long-read bulk RNA sequencing
identifies isoforms comprehensively but lacks single-cell specificity. In this study, we employ
single-cell nanopore sequencing of full-length mRNAs across 27 glioblastoma
patients, yielding approximately 210,000 single-cell transcriptomes.
Integrating these results with paired short-read scRNA-seq
allowed precise cell-type identification and detection of cell-type-specific
isoforms. This approach enabled the discovery of cell-type-specific novel
splicing events, including new combinations of known splice sites and
undiscovered donor and acceptor sites, revealing a remarkable level of
transcriptomic diversity within glioblastoma. This extensive single-cell
analysis provides unprecedented insights into the transcriptional landscape
of glioblastoma, identifying novel markers of transcriptional differentiation
and potential therapeutic targets at single-cell resolution, advancing our
understanding of splicing-driven glioblastoma heterogeneity and tumor
progression. |
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Tay, Wan Jing National University Hospital
(Singapore) Clinical and Molecular Divergences
in Diffuse Midline Glioma, H3 K27-Altered: A Comparative Study of Survival Outcomes
in Adults and Children Background: Diffuse midline glioma, H3
K27-altered (DMG), affects both adults and children. Previous studies
reported conflicting data regarding survival differences between adults and
children with DMG. This study compared clinical characteristics, molecular
profiles, and survival outcomes between adult and paediatric
patients with DMG. Methods: A pooled analysis of 1,498 patients
(521 adults, 977 children) diagnosed with DMG was conducted using published
data. The individual patient time-to-event data for overall survival (OS),
defined as the time from diagnosis to death from any cause, were
reconstructed from published Kaplan-Meier curves. Descriptive statistics were
used to summarize the characteristics and molecular profiles. Cox regression
was performed to compare OS between adults and children. Subgroup analyses
were performed to evaluate potential factors that could modify the effect on
OS between the two groups. Results: Brainstem involvement was less
frequent in adults than in children (26% vs. 79%). Adults had higher rates of
PPM1D (37% vs. 13%), FGFR1 mutations (17% vs. 6%), and NF1 mutations (35% vs.
12%) and lower rates of ACVR1 mutations (5% vs. 24%) than children. Adults
had longer OS (median 16.2 months vs. 11.4 months, Hazard Ratio (HR) 0.52
(95% CI 0.46 – 0.59), p < 0.0001) than children. Subgroup analyses showed
that brainstem involvement was a potential modifier on the effect of OS
between adults and children (Brainstem: HR 0.47 (95% CI 0.33 – 0.66) vs No
brainstem: HR 0.91 (95% CI 0.67 – 1.25), interaction P = 0.006). ATRX, NF1,
TP53, and PPM1D mutations were not effect modifiers. Conclusion: The involvement of the brainstem,
mutations in PPM1D, FGFR1, NF1, and ACVR1 genes differed between adults and
children with DMG. Overall, adults had improved OS compared to children,
especially in those with brainstem involvement. These findings highlight the
need to further investigate the underlying biological differences in DMG
between age groups. |
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Varlet, Pascale GHU Paris-Psychiatrie
et Neurosciences, Sainte-Anne Hospital Methylation-based RTK2A and RTK2B
diffuse pediatric high-grade gliomas present distinct radiological and histomolecular features Diffuse pediatric-type high-grade
gliomas (pedHGG), H3- and IDH-wildtype, encompass
three main DNA-methylation-based subtypes: pedHGG-MYCN,
pedHGG-RTK1A/B/C, and pedHGG-RTK2A/B. Since their first description in 2017
tumors of pedHGG-RTK2A/B have not been comprehensively characterized and
their clinical significance is unknown. In a recent series of pedHGG with a gliomatosis cerebri (GC) growth pattern, an
increased incidence of pedHGG-RTK2A/B (n=18) was observed. We added 14
epigenetically defined pedHGG-RTK2A/B tumors to this GC series and provided
centrally reviewed radiological, histological, and molecular
characterization. The final cohort of 32 pedHGG-RTK2A/B tumors consisted of
25 pedHGG-RTK2A (78%) and seven pedHGG-RTK2B (22%). The median age was 11.6
years (range, 4–17) with a median overall survival of 16.0 months (range
10.9–28.2). Seven of 11 added cases with imaging available showed a GC
phenotype at diagnosis or follow-up. PedHGG-RTK2B tumors exhibited frequent bithalamic involvement (6/7, 86%). Central neuropathology
review confirmed a diffuse glial neoplasm in all tumors with prominent
angiocentric features in both subclasses. Most tumors (24/27 with available
data, 89%) diffusely expressed EGFR with focal angiocentric enhancement.
PedHGG-RTK2A tumors lacked OLIG2 expression, whereas 43% (3/7) of
pedHGG-RTK2B expressed this glial transcription factor. ATRX loss occurred in
3/6 pedHGG-RTK2B samples with available data (50%). DNA sequencing
(pedHGG-RTK2A: n=18, pedHGG-RTK2B: n=5) found EGFR alterations (15/23, 65%;
predominantly point mutations) in both subclasses. Mutations in BCOR (14/18,
78%), SETD2 (7/18, 39%), and hTERT promoter (7/18, 39%) occurred exclusively
in pedHGG-RTK2A tumors, while pedHGG-RTK2B tumors were enriched for TP53
alterations (4/5, 80%). In conclusion, pedHGG-RTK2A/B tumors revealed a
highly diffuse-infiltrating tumor with specific radiological and histomolecular features. By comprehensive characterizing
specific methylation-based tumors, the chance to facilitate specific and
effective therapy concepts for these so far detrimental tumors increases. |
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Vella, Viviana University of Sussex Kinome-Wide Synthetic Lethal Screen Identifies PANK4 as a
Modulator of Temozolomide Resistance in Glioblastoma Background and Aims: Temozolomide (TMZ) represents the
cornerstone of glioblastoma (GBM) therapy. However, acquisition of resistance
limits its therapeutic potential. The human kinome
is an undisputable source of druggable targets, still, current knowledge
remains confined to a limited fraction of it, with a multitude of
under-investigated proteins yet to be characterised.
In this study, we aimed to uncover synthetic lethal partners of TMZ in GBM
that could enhance the effect of TMZ, thus improving TMZ therapy response. Methods: A kinome-wide
RNAi screen was performed in a TMZ-resistant GBM cell model. A panel of
several TMZ-resistant and patient-derived GBM cell lines was implemented for
in vitro validation of our findings through several phenotypic assays. In
vivo TMZ-resistant GBM xenografts and immunohistochemical (IHC) analysis of
IDH-wildtype GBM specimens were employed to assess the clinical relevance of
our findings. Tandem Mass Tag (TMT)-based quantitative proteomic studies were
undertaken to elucidate the underlying mechanisms. Results: Following a kinome-wide
RNAi screen, we identified pantothenate kinase 4 (PANK4) as a modulator of
TMZ resistance in GBM and demonstrated that its depletion enhances the effect
of TMZ, therefore improving the response to TMZ therapy. Validation of PANK4
across various TMZ-resistant GBM cell models and patient-derived GBM cell
lines, immunohistochemical (IHC) analysis of IDH-wildtype GBM specimens, as
well as in vivo studies, corroborated the potential translational
significance of these findings. We also provided evidence that PANK4
expression is induced in response to TMZ treatment and increased PANK4 levels
are associated with a worse clinical outcome. Moreover, by employing a Tandem
Mass Tag (TMT)-based quantitative proteomic approach we revealed that PANK4
abrogation leads to a significant downregulation of numerous proteins with
central roles in cellular detoxification and cellular response to oxidative
stress. Furthermore, we showed that the ability of PANK4 to modulate
sensitivity to TMZ treatment and intracellular reactive oxygen species (ROS)
accumulation is dependent on its phosphatase activity. Mechanistically, as
cells undergo genotoxic stress during TMZ exposure, PANK4 depletion
represents a crucial event that can lead to accumulation of intracellular ROS
and subsequent cell death. Conclusion: Collectively, our study provides
novel insights into chemoresistance in GBM and unveils a previously
unreported role for PANK4 in mediating therapeutic resistance to TMZ in GBM. |
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Wadekar, Ashish All India Institute of Medical Sciences, New Delhi Evaluating the efficacy of Hip1R, Vimentin, and H3K27Me3 as
surrogate markers for 1p/19q codeletion in oligodendrogliomas Assessment of the chromosomal 1p/19q status is essential to
distinguish between IDH mutant astrocytoma and oligodendroglioma. Genetic
analyses, however, are expensive, time-consuming, and not widely accessible.
Immunohistochemical loss of ATRX is currently the only established surrogate
marker for a non-1p/19q-codeleted genotype. To find cost-effective approaches
and improve risk stratification, we aimed to investigate markers such as
Vimentin, H3K27me3, and HIP1R. We aimed to assess the immunohistochemical expression HIP1R,
Vimentin, and H3K27me3 as surrogate markers in predicting 1p/19q status in
oligodendrogliomas. A total of 182 adult-type diffuse glioma patients were analyzed
for IDH 1 R132H, ATRX, P 53, MIB1-LI, HIP1R, Vimentin, and H3K27me3
expression using immunohistochemistry. 1p/19q co-deletion was assessed by
FISH assay. IDH sequencing was performed in IDH 1 R132H negative cases. Histomorphological and molecular classification of these
gliomas was performed according to WHO 2021 CNS5 classification. In this study 102 IDH-mutant oligodendrogliomas, 44 IDH-mutant astrocytomas, and 36 IDH-wild-type glioblastomas,
exhibited distinct patterns of the IHC markers. In oligodendrogliomas, HIP1R
showed either homogeneous or homogenous and mosaic staining, Vimentin was
negative and H3K27me3 was lost in all cases. In IDH mutant astrocytomas and glioblastomas, HIP1R was predominantly
mosaic, Vimentin was widely positive, and H3K27me3 was variable. Combining
these markers, especially positivity of HIP1R, negative Vimentin, and
complete loss of H3K27me3, achieved perfect diagnostic accuracy, making them
highly reliable for differentiating oligodendrogliomas from astrocytomas, and glioblastomas. The study demonstrates that the combination of the three
immunohistochemical markers HIP1R, Vimentin, and H3K27me3 can accurately
predict 1p/19q co-deletion status in IDH-mutant gliomas. This method offers a
reliable, robust and cost-effective alternative to complex techniques like
FISH. |
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Wang, Da-Peng Ruijin Hospital, Shanghai JiaoTong
University The m6A reader HNRNPC mediates BRD4
stabilization enhancing cabergoline efficacy in prolactinomas Objective: Pituitary adenomas (PAs), common
intracranial benign tumors, include hormone-secreting functioning tumors and
non-functioning pituitary adenomas. Prolactinomas (PRLomas)
are usually treated with dopamine agonists (DAs), including cabergoline
(CAB). Unfortunately, drug resistance occurs in approximately 20% of
patients. Bromodomain protein 4 (BRD4), a transcriptional regulator that can
be methylated by N6-methyladenosine (m6A), is involved in tumor growth and
drug resistance, but its role in PRLomas is still
unclear. Methods: Herein, the expression of BRD4 mRNA
and protein was examined in PRLoma samples. The
effect of silencing RNA against BRD4 (SiRNA-BRD4), treatment with CAB and
combined treatment with both CAB and SiRNA-BRD4 on cell growth, apoptosis and
PRL secretion were tested in MMQ and CH3 cells. Post-treatment mitochondrial
oxidative stress and untargeted metabolomics were also evaluated. HNRNPC,
BRD4, Scg2, DRD2 and apoptosis proteins were assessed by western blot. BRD4
methylation was investigated using m6A IP (MeRIP),
dual-luciferase reporter and Actinomycin D assays. JQ-1, a BRD4 inhibitor,
and CAB were used in eutopic and ectopic PRLoma animals. Results: BRD4 was highly expressed in PRLomas especially in patients with DA resistance, and it
is positively related to drug resistance related genes, including TP53,
SSTR2, PXN and KBTBD6. Inhibition of BRD4 promoted mitochondrial oxidative
stress and dysbolism, apoptosis and PRL decline in
cells. Compared to monotherapy, polytherapy had superior effects on
anti-tumor growth and PRL suppression. SiRNA-BRD4 inhibited Scg2 expression
but activated the DRD2 protein. Co-IP experiments confirmed the interactions
between Scg2 and BRD4, Scg2 and DRD2. Meanwhile, the m6A reader HNRNPC
methylated the 3’-untranslated region (UTR) but not the coding sequence (CDS)
of BRD4, regulating its transcription. Conclusion: Inhibition of BRD4 can enhance the
anti-tumor effects of CAB with a synergistic effect against PRLomas, which may be a viable therapeutic approach for
DA-resistant patients. |
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Wang, Da-Peng Fudan University NKX2-5 as a promising prognostic
indicator linked to tumor immune microenvironment in gliomas Background: NK2 homeobox 5 (NKX2-5) is a
homeobox-containing transcription factor that exhibits differential
expression in various cancers. However, its prognostic significance,
influence on tumor immune infiltration, and potential mechanisms in glioma
development remain elusive. Methods: We first conducted an analysis
using data from The Cancer Genome Atlas (TCGA) and corroborated our findings
with data from the Chinese Glioma Genome Atlas (CGGA). Univariate and
multivariate evaluation of NKX2-5 in glioma prognosis. The UALCAN and MethHC databases were used to analyze the correlation
between NKX2-5 and methylation, as well as methylation prognosis. The ssGSEA algorithm was further applied to analyze the
association of NKX2-5 with immune cell infiltration, immune checkpoints, and
drug sensitivity. GO and KEGG enrichment analyses were conducted to explore
the potential biological functions and signaling pathways of the top 50
co-expressed genes with NKX2-5 in the TCGA database. RT-qPCR and
immunohistochemistry experiments were used to detect the mRNA and protein
expression levels of NKX2-5 in clinical tissue samples. MTS, EDU, Transwell, and wound healing assays were performed to
evaluate the effects of NKX2-5 knockdown on the proliferation, invasion, and
migration of glioma cells. Hoechst 33342 staining and flow cytometry were
used to assess the effect of NKX2-5 deficiency on TMZ sensitivity in glioma
cells, and ELISA was employed to measure IL6 and TNF expression levels. Results: Our univariate and multivariate Cox
analyses confirmed that increased NKX2-5 mRNA levels are an independent
predictor of poor prognosis. Glioma progression and stage were negatively
associated with NKX2-5 promoter methylation. Interestingly, different
methylation sites on the NKX2-5 gene were associated with distinct prognostic
outcomes of glioma patients. Furthermore, High expression of NKX2-5 in glioma
is positively correlated with macrophage infiltration and negatively
correlated with CD8+ T cell infiltration. In addition, NKX2-5 may influence
the tumor immune microenvironment and subsequently impact immune checkpoint
activity, immune checkpoint blockade (ICB) therapy, and chemotherapy
responses. Moreover, co-expression genes associated with NKX2-5 primarily
function as extracellular matrix structural constituents and enzyme inhibitors,
the signaling pathway involved in cell cycle, lysosome, and phagosome
processes. Clinically, we confirmed the NKX2-5 is highly expressed in glioma
and knockdown of NKX2-5 significantly reduced tumor proliferation and
aggressive behavior. Additionally, NKX2-5 knockdown enhanced the sensitivity
of glioma cells to the chemotherapeutic drug-temozolomide. Conclusion Our findings suggest for
the first time that NKX2-5 is a promising prognostic indicator for gliomas
and a critical player in tumor immune microenvironment. These findings
highlight its potential as a novel therapeutic target for future
immunotherapy interventions. |
|
Wei, Yanfei Henan Academy of Sciences Stalled oligodendrocyte
differentiation in IDH‑mutant
gliomas Roughly 50% of adult gliomas harbor
isocitrate dehydrogenase (IDH) mutations. According to the 2021 WHO
classification guideline, these gliomas are diagnosed as astrocytomas,
harboring no 1p19q co-deletion, or oligodendrogliomas, harboring 1p19q
co-deletion. Recent studies report that IDH-mutant gliomas share a common
developmental hierarchy. However, the neural lineages and differentiation
stages in IDH-mutant gliomas remain inadequately characterized. Using bulk
transcriptomes and single-cell transcriptomes, we identified genes enriched
in IDH-mutant gliomas with or without 1p19q co-deletion, we also assessed the
expression pattern of stage-specific signatures and key regulators of
oligodendrocyte lineage differentiation. We compared the expression of oligodendrocyte
lineage stage-specific markers between quiescent and proliferating malignant
single cells. The gene expression profiles were validated using RNAscope analysis and myelin staining and were further
substantiated using data of DNA methylation and single-cell ATAC-seq. As a
control, we assessed the expression pattern of astrocyte lineage markers.
Genes concordantly enriched in both subtypes of IDH-mutant gliomas are
upregulated in oligodendrocyte progenitor cells (OPC). Signatures of early
stages of oligodendrocyte lineage and key regulators of OPC specification and
maintenance are enriched in all IDH-mutant gliomas. In contrast, signature of
myelin-forming oligodendrocytes, myelination regulators, and myelin
components are significantly down-regulated or absent in IDH-mutant gliomas.
Further, single-cell transcriptomes of IDH-mutant gliomas are similar to OPC
and differentiation-committed oligodendrocyte progenitors, but not to
myelinating oligodendrocyte. Most IDH-mutant glioma cells are quiescent; quiescent
cells and proliferating cells resemble the same differentiation stage of
oligodendrocyte lineage. Mirroring the gene expression profiles along the
oligodendrocyte lineage, analyses of DNA methylation and single-cell ATAC-seq
data demonstrate that genes of myelination regulators and myelin components
are hypermethylated and show inaccessible chromatin status, whereas
regulators of OPC specification and maintenance are hypomethylated and show
open chromatin status. Markers of astrocyte precursors are not enriched in
IDH-mutant gliomas. These findings show that despite differences in clinical
manifestation and genomic alterations, all IDH- mutant gliomas resemble early
stages of oligodendrocyte lineage and are stalled in oligodendrocyte
differentiation due to blocked myelination program. These findings provide a
framework to accommodate biological features and therapy development for
IDH-mutant gliomas (Genome Medicine (2023) 15:24). |
|
Wong, Cheuk Lun The University of Hong Kong Yes-associated protein (YAP)
modulation in targeting temozolomide chemoresistance in glioblastoma The Hippo signaling pathway is
often dysregulated in cancer. The study aims to investigate whether the Hippo
signaling pathway contributes to temozolomide chemoresistance in glioblastoma,
and to validate whether the combinational treatment of temozolomide and YAP
inhibitor is effective against glioblastoma. Temozolomide-sensitive and
temozolomide-resistant U87 and U251 GBM human cell lines were previously
established. Temozolomide-resistant cells were maintained in low-dose
temozolomide. Here, we investigate whether Hippo signaling contributes to
temozolomide chemoresistance. By modulating the Hippo signaling pathway
through short-hairpin YAP gene knockdown, the outcome on GBM cancer
phenotypes was studied by cell assays (in vitro) and mouse orthotopic
xenograft (in vivo). Finally, the therapeutic effect of YAP inhibitor is
studied in vitro and in vivo respectively. Our data showed that YAP is
overexpressed in temozolomide-resistant glioblastoma cells. Nuclear
translocation of YAP (which becomes its active state) is increased in
temozolomide-resistant cells. In fact, YAP inhibition demonstrates reduced
cell viability and higher susceptibility to temozolomide. Mice with tumor
injection of the YAP knockdown cells have significantly reduced tumor size.
Similarly, YAP inhibitor resensitizes
temozolomide-resistant cells to temozolomide. Mechanistically, YAP knockdown
reduces cell stemness, which in turn attenuates temozolomide chemoresistance.
In conclusion, our data suggests dysregulated Hippo signaling pathway
contributes to temozolomide chemoresistance, thus targeting the Hippo
signaling pathway resensitizes GBM cells to
temozolomide. This research provides pre-clinical evidence on using YAP
inhibitor for combinational therapy with temozolomide for the treatment of
glioblastoma. |
|
Wong, Ka Ho LKS Faculty of Medicine, The
University of Hong Kong Immunogenicity Rescue of Tumor
Microenvironment through the Manipulation of Low-density Lipoprotein
Receptor-related Protein 1 in Tumor-Associated Macrophages in Glioblastoma Purpose: Glioblastoma (GBM) is the most
common malignant primary brain tumor. The tumor microenvironment (TME) of GBM
possesses a myeloid-dominant nature with T-cell scarcity, rendering
traditional lymphoid-targeting immunotherapy ineffective. Myeloid-modifying
treatment targeting tumor-associated macrophages (TAMs), historically
described as immunosuppressive, is emerging as a promising alternative. In
this study, we discovered a novel subpopulation of Low-density lipoprotein
receptor-related protein 1 (LRP1)-high TAMs that exhibit anti-tumoral
characteristics and showcased the therapeutic potential of modifying the LRP1
eat-me pathway in glioblastoma. Methods: Single-cell RNA sequencing (scRNA-seq) and spatial transcriptomic analysis was
performed on clinical samples of glioblastoma patients. CX3CR1+/GFP and
myeloid-depleted (CD11b-DTR) xenografts were utilized to assess in vivo
macrophage characteristics and monitor tumor growth. Specifically, TAMs were
subjected to in vivo LRP1 knockdown/overexpression through
macrophage-specific AAV. Primary bone marrow-derived macrophages (BMDM) were
generated from mouse bone marrow stromal cells, subjected to lentiviral
knockdown/overexpression of LRP1, and subsequently used to reconstitute the
macrophage environment of CD11b-DTR mice after myeloid depletion using
diphtheria toxin. Tumors were collected from the xenografts for downstream
analysis, including scRNA-seq, flow cytometry,
immunofluorescence and immunohistochemical imaging, and protein/mRNA
expression analysis. For in vitro experiments, knockdown of LRP1 and
co-culture experiments with glioblastoma cells were performed with BMDM,
human monocytic cell lines (U937, THP1), mouse macrophage cell line (RAW
246.7) and primary peripheral blood macrophages (PbMC)
to assess in vitro phagocytosis and migration abilities. Results: scRNA-seq of glioblastoma patient samples reveals that LRP1
is highly expressed in pro-inflammatory (anti-tumoral) TAMs compared to
anti-inflammatory (pro-tumoral) TAMs. LRP1-high TAMs demonstrate highly
enriched anti-tumoral signaling pathways including phagocytosis and
migration. Moreover, higher LRP1 expression in the TAMs is correlated with
better patient survival. Spatial transcriptomic analysis identified the
LRP1-calreticulin (one of the ""eat-me"" signaling
pathways) as the major mode of interaction between LRP1-high TAMs and GBM
tumor cells. In vivo xenografts demonstrated that AAV-mediated modification
of LRP1 on TAMs leads to a stepwise impediment of tumor growth and
enhancement of survival from LRP1 knockdown to control to overexpression. scRNA-seq analysis of tumors CD11b-DTR myeloid-depleted
mice with BMDM reconstitution shows a more immunosuppressive TME landscape
with LRP1-knockdown BMDM, and a more immune-activating TME landscape with
LRP1-overexpressed BMDM, compared to control. In vitro co-culture experiments
show that phagocytic rate is significantly reduced after LRP1 knockdown in
macrophages. In addition, we identified vitamin D receptor (VDR) possesses
transcriptomic control over LRP1 promoter through chromatin
immunoprecipitation analysis, with LRP1 expression showing calcitriol (active
vitamin D) dose-dependent increase on both mRNA and protein levels. Oral
calcitriol treatment effectively promotes macrophage infiltration and
phagocytosis in xenograft models. Conclusions: We hereby uncovered a novel
LRP1-high subpopulation of TAMs showcasing strong anti-tumoral potentials,
which is in stark contrast to the traditional notion of TAMs being
immunosuppressive. We demonstrate the pivotal role of LRP1 in governing TAMs' anti-tumoral abilities in GBM. We also illustrated
the therapeutic potential of myeloid-modulating treatments in GBM through
LRP1 pathway enhancement using calcitriol treatment. |
|
Xi, Shaoyan Sun Yat-sen
University Cancer Center Comprehensive Analysis of
Supratentorial Ependymoma, ZFTA-Fusion Positive with non-RELA Partners The definition of ependymoma has
moved from purely histopathological features to integrated diagnosis, relying
heavily on molecular information. The 2021 World Health Organization (WHO)
classification of central nervous system (CNS) tumors defines two molecular
subtypes of supratentorial ependymoma (ST-EPN): one with zinc finger
translocation-associated protein (ZFTA) fusion and another with
yes-associated protein 1 (YAP1) fusion. The former has been previously named
“ST-EPN, RELA-fusion positive” because ZFTA-RELA fusion was detected in most cases.
However, ZTFA is now considered more representative than RELA because it may
be fused with other partners. Here, we present ten additional cases of
ST-EPN, ZFTA-fusion positive with non-RELA partners such as mastermind-like
protein 2 (MAML2) (n=2), nuclear receptor coactivator 2 (NCOA2) (n=4),
contactin-5 (CNTN5) (n=1), terminal uridylyl transferase 1 (TUT1) (n=1), synaptotagmin 12 (SYT12) (n=1), and transmembrane
O-methyltransferase (LRTOMT) (n=1). Various histological characteristics were
observed, including classic EPN-like, sarcoma-like, metastatic cancer-like,
embryonal tumor-like, and even chordoma-like. These histological patterns
brought out many differential diagnoses without next generational sequencing
(NGS). DNA methylation profiling clustered the cases with the reference
“EPN_ZFTA”. Together with the existing literature, we summarized 30 cases of
ST-EPN, ZFTA-fusion positive with non-RELA partners. This tumor predominantly
affected children and young adults without sexual predilection. Compared to
ST-EPN with typical ZTFA-RELA fusion, tumors carrying non-RELA fusion
partners showed a significantly better prognosis. Our study further expanded
the morphologic and molecular genetic spectrum of ST-EPN, ZFTA-Fusion
positive with non-RELA partners. |
|
Yang, Ming Cancer Hospital China Academy of
Medical Sciences, National Cancer Centre China The Safety and Efficacy of a
cocktail therapy of Zanubrutinib, rituximab and High Dose- Methotrexate in
treating Patients with Newly Diagnosed Primary Central Nervous System Lymphoma Primary central nervous system
lymphoma (PCNSL) is an aggressive non-Hodgkin lymphoma presenting with
disease limited to the brain, spinal cord, leptomeninges, cerebrospinal fluid
(CSF) and eyes. High-dose methotrexate (HD-MTX) chemotherapy is the preferred
standard induction treatment regimen. We retrospectively studied the safety
and efficacy of a combined cocktail therapy of Zanubrutinib, rituximab and
HD-MTX as the induction therapy for PCNSL. The study enrolled a total of 99
patients with PCNSL treated in the Cancer Hospital Chinese Academy of Medical
Science between 1st January 2021 and 31st December 2023. Among them, 18
patients received the cocktail therapy with rituximab, methotrexate and
Zanubrutinib as the induction therapy. The safety and efficacy data were
collected, including treatment response with follow up data, side effects,
and prognosis. It has shown a promising result of the combined cocktail
therapy as an induction treatment for PCNSL, which showed a favorable
curative efficiency and milder adverse reactions. Moreover, a genetic
analysis was performed. We analyzed the genetic profile associated with poor
response and poor prognosis to the therapy. A few genetic footprints were
identified associated with the poor response to the cocktail therapy. Thus,
we propose the combined cocktail therapy of Zanubrutinib, rituximab and
HD-MTX as the induction therapy for PCNSL is potentially a safe and effective
option in treatment. And genetic screen may help to select the potential poor
responders. |
|
Yang, Ming Cancer Hospital China Academy of
Medical Sciences, National Cancer Centre China Spinal Cord High Grade Glioma: A
case series and review of the literature In this paper, we present 2 cases
of spinal HGG, one with diffuse midline glioma (DMG) and another patient with
high grade astrocytoma, looking specifically into their histology subtypes,
progress and treatment outcomes, along with a literature review. Case study 1, a 35-year-old female
presented with one month of neck pain after carrying heavy items. On
examination she had left hemiparesis. MRI cervical spine revealed a large
solitary enhancing intramedullary lesion from C2 to C5. She underwent C2-C6
laminectomy and debulking of the lesion. Histology showed high grade diffuse
glioma, IDH wild type, with loss of staining for H3K27me3 and diffusely
positive staining for H3K27M. Post operatively, she underwent fractionated
radiotherapy (RT) with concurrent temozolomide (TMZ). Lomustine
and bevacizumab was initiated, for which she is currently still receiving. Case Study 2 A 49-year-old male presented with
bilateral lower limb numbness, proprioceptive loss and urinary incontinence.
MRI spine showed a T10–12 intramedullary lesion and he underwent a T11
laminectomy and biopsy. Histology returned as high grade astrocytoma,
IDH1R132H negative, H3K27M negative. He underwent concurrent TMZ and RT with
bevacizumab for 3 months, followed by adjuvant TMZ and Bevacizumab (BEV) for
1 year. However he developed worsening lower limb numbness and weakness and
was switched to Lomustine/Bevacizumab for a year.
Then his chemotherapy was changed to carboplatin and etoposide.This
patient was alive at the 4 year follow up, however he developed extensive
leptomeningeal disease with intracranial lesions and is currently undergoing
palliative RT. Literature Review Evidence for the treatment of
spinal HGG remains scarce and limited to mainly case reports. Most therapies
are extrapolated from intracranial HGG studies, the standard of care being
maximal safe resection followed by concurrent RT and TMZ for 6 weeks, then
adjuvant TMZ for 6 months. In one of the largest case series
on primary spinal HGG of 29 patients, Upadhyay et al found no impact of
concurrent chemotherapy on survival, with Temozolomide having limited
evidence of benefit. However Temozolomide and Bevacizumab in particular have
been shown to improve survival in retrospective series in recurrent HGGs,
with Bevacizumab thought to decrease peritumoral oedema and mass effect. Outside
of TMZ, there are three drugs and one device which are FDA- approved for the
treatment of recurrent HGGs, namely Lomustine, Carmustine, Bevacizumab and tumour
treating fields. Conversely, adjuvant RT has been shown to improve survival
in patient with WHO grade 2-4 infiltrative astrocytoma, with the rate of
local failure in spinal HGG of only 55% compared to over 80% for intracranial
glioblastomas in the Stupp trial Recent trials have shown that convection
enhanced delivery, blood-brain barrier opening, and immunotherapy are
tolerated well by patients with H3K27M-mutated diffuse midline glioma, paving
the way for future trials with novel agents and targeted therapies. Conclusion Prognosis of spinal HGG patient
remains poor despite current standard of care with maximal safe resection and
adjuvant chemoradiation. A better understanding of the molecular drivers of
spinal HGGs and more clinical trials are needed to develop more effective
treatment options. |
|
Yao, Shun The First Affiliated Hospital, Sun
Yat-sen University Leveraging Generative AI for
MRI-to-CT Translation: Advancements and Applications in Neurosurgical
Planning Objective: Computed tomography (CT) and
magnetic resonance imaging (MRI) provide complementary insights for neurosurgical
planning. While both modalities are valuable, obtaining both scans increases
costs and exposes patients to additional ionizing radiation following an MRI.
Cross-modal generation of high-resolution CT images from MRI data offers a
promising alternative. This study aimed to develop deep learning-based
frameworks, including a modified conditional generative adversarial network (cGAN) and a novel mask-guided dual network based on GAN
architecture (MGDGAN), to synthesize CT (sCT)
images from MRI scans. Methods: Pre-operative paired CT and
contrast-enhanced MRI scans were collected from patients with conditions
including meningiomas, pituitary tumors, vestibular schwannomas, and
cerebrovascular diseases. The CT and MRI images were processed for denoising,
field correction, and co-registration. MRI data were then used as input for
the generative-AI frameworks, which generated sCT
images. The accuracy of these synthetic images was assessed through
quantitative similarity metrics and by evaluating clinical features,
including measurements of the sella, internal
auditory canal (IAC), and aeration patterns in the mastoid, clinoid, and
sphenoid regions. Results: A total of 92,981 paired CT/MRI
images from 80 patients were used for training and testing, with an
additional 10,068 paired images from 10 patients serving as an external
validation set. The sCT images successfully
reconstructed the bony skull base and convexity with high accuracy. The
median relative error in sella and IAC measurements
between synthetic and real CT (rCT) scans was 6%,
though there was greater variability in IAC reconstructions. Aeration in the
mastoid, clinoid, and sphenoid regions was generally well-represented,
although finer air cell septations showed some heterogeneity. Performance
varied by pathology, with the most significant challenges in cases of
meningiomas with intratumoral calcifications or calvarial invasion. Additionally, experimental results
indicated that MGDGAN achieved high accuracy in generating fine bone
structures, brain tissues, and cerebral lesions, producing images visually
closer to rCT scans. Quantitative evaluations
showed that MGDGAN outperformed other state-of-the-art methods, including CycleGAN, Pix2Pix, ECNN, cGAN9, APS, and ResViT across multiple datasets. Conclusions: The proposed generative AI
frameworks, cGAN and MGDGAN, for MRI-to-CT
translation have demonstrated significant potential for various neurosurgical
applications, particularly as an adjunct tool for pre-operative planning of
skull base surgeries. Further refinement of GAN variants performance for
specific anatomical regions may further enhance its clinical utility. |
|
"Yao, Shun The First Affiliated Hospital, Sun
Yat-sen University MRI Atlas of Meningioma Spatial
Distribution across Molecularly Integrated Grades Background: It is recognized that the site of
meningioma origins across the meninges correlates with distinct symptoms,
tissue histopathology subtypes, and surgical risks. However, pinpointing the
exact location of the tumor's origin can be challenging, particularly in
large tumors or those that span different brain areas. We employed an
unbiased lesion mapping method to identify where meningiomas originate and
how this relates to their biological characteristics and grading. Methods: We analyzed MRI scans, WHO grades,
and a novel grading system called Integrated Grade (IG), which is based on
genetic information, from 881 adult patients with confirmed intracranial
meningiomas at a tertiary academic medical center. We utilized a semi-automated
method to contour the tumors on pre-operative MRI scans and subsequently
created a frequency atlas of meningioma locations using voxel-based lesion
mapping (VBLM). This atlas illustrated the frequency of tumor occurrence in
different areas and was compared to the tumors' origins. Results: Among the 881 patients (average age
57, 68.8% female), 589 had WHO grade 1 tumors (66.8%), 265 had WHO grade 2
(30.1%), and 27 had WHO grade 3 (3.1%). After reclassification based on
molecular criteria, 585 were IG-1 (66.3%), 160 were IG-2 (18.2%), and 136
were IG-3 (15.5%). Less aggressive tumors were predominantly found in the
middle of skull base and its surroundings. More aggressive meningiomas were
more frequently observed in the falcine/parasagittal
area and the right sphenoid region. When examining the molecular Integrated
Grade, the clustering of tumors in certain areas was even more pronounced
than when considering the WHO grade. WHO grade 2 tumors were distributed
across IG 1-3, and their locations also varied accordingly. Conclusion: By employing the unbiased VBLM
technique, we demonstrated that meningiomas exhibit a preference for specific
areas, with certain areas being associated with more aggressive tumors.
Different tumor distribution patterns were observed when considering both
histopathological type and molecular grade. The molecular grade was
associated with more distinct clusters of meningiomas, supporting the
hypothesis that there is an association between the phenotype and genotypes
of meningiomas. |
|
Zhang, Qp The 6th Affiliated Hospital of
Shenzhen University Medical School Brain cancer stem cells exosomes
derived NamiRNA-enhancer network of miR-151a-3p
activates PDE4D to promote Progression of glioma Purpose: Glioblastoma (GBM) is the most
aggressive form of brain cancer, with a median survival time of less than 15
months despite multimodal treatment approaches. Brain tumor stem cells
(BTSCs) contribute significantly to tumor recurrence and resistance to
therapy. The molecular mechanisms by which BTSCs-derived exosomes influence
glioma invasion and migration, particularly under hypoxic conditions, remain
elusive. How miRNAs in the hypoxic BTSCs (H-BTSCs)-derived exosomes promoting
the progression of glioma is currently unclear. Thus, studying these in depth
would be beneficial in understanding their roles and mechanisms in different
diseases. Methods: We conducted a comprehensive study
involving high-throughput sequencing to identify differentially expressed
miRNAs in exosomes derived from hypoxic BTSCs (H-BTSCs). We then employed
Chip, dual luciferase reporter assays et al found that a NamiRNA-enhancer
network of miR-151a-3p have the direct interaction between the
phosphodiesterase 4D (PDE4D) protein. The study included in vitro assays
using glioma cell lines, patient-derived xenograft (PDX) models, and
biochemical analyses to assess the functional role of miR-151a-3p and PDE4D
in glioma progression. Results: Our findings revealed a significant
increase in miR-151a-3p levels under hypoxic conditions, which directly
interacted with the PDE4D enhancer to activate the focal adhesion signaling
pathway. miR-151a-3p upregulated PDE4D expression in glioma cells by
enhancing the PDE4D promoter activity, forming a positive feedback loop that
amplified glioma development. In vivo studies confirmed that H-BTSCs-derived
exosomes rich in miR-151a-3p significantly enhanced tumor growth, invasion,
and migration in orthotopic xenograft models. Conclusion: This study identifies a novel
NamiR-151a-3p/PDE4D/focal adhesion kinase signaling axis that plays a
critical role in the progression of glioblastoma. Our findings not only
reveal a key mechanism by which hypoxic BTSCs-derived exosomes promote glioma
invasion and migration but also provide a potential therapeutic target for
the treatment of GBM. The identification of miR-151a-3p and PDE4D as central
players in this process offers a new avenue for diagnostic and therapeutic
intervention in glioblastoma. |
|
Zheng, Danfeng Third Hospital, School of Basic Medical Sciences, Peking
University Health Science Center A preliminary study on the clinical and pathological factors
related to postoperative recurrence in patients with meningioma Background: Meningiomas are the most common primary tumor of the central
nervous system, which are usually benign tumors that can be cured by surgical
resection, but some patients relapse even after total surgical resection. How
to solve the recurrence of meningiomas has always been a difficult problem in
clinical work. Therefore, this study focused on exploring the clinical and
pathological characteristics associated with meningioma recurrence after
surgery, and screened the candidate gene KCNMA1 through the database to
predict meningioma recurrence, and initially explored its protein expression
in meningiomas and the molecular mechanism of its down-regulation in
recurrent meningiomas.
The clinical and pathological baseline data of 175 meningioma
patients were collected in this study. Univariate and multivariate binary
logistic regression were used to analyze the factors that influence patient
recurrence. The differentially expressed gene KCNMA1 was screen out related
to meningioma recurrence, and then KCNMA1 immunohistochemical staining was
performed in 82 meningioma samples to explore its protein expression level in
meningioma. Bisulfite PCR based on next-generation sequencing was used to
detect specific DNA point mutations and promoter methylation levels of the
KCNMA1 gene in paraffin-embedded meningioma tissue specimens. Results: 1) recurrence occurred in 39 of 175 patients with meningiomas
(recurrence rate 22.3%), including 23 of 144 patients with WHO grade 1
meningiomas (recurrence rate 16.0%). In the multivariate regression analysis,
mitotic count ≥ 3/10 HPF (equivalent to 1.6mm2), WHO high grade and
maximum tumor diameter ≥ 5 cm were prognostic factors for meningioma
recurrence (P < 0.05). Moreover, in patients with WHO grade 1 meningiomas,
the risk of recurrence of tumor with mitotic count ≥ 3/10 HPF and
maximum diameter ≥ 5 cm is higher than that of overall meningiomas. 2)
The expression level of KCNMA1 protein was down-regulated in recurrent
meningiomas, and was an independent prognostic factor for recurrence of
meningiomas included in multivariate analysis(overall: OR, 0.678, 95%CI,
0.570-0.807, P < 0.001; WHO grade 1: OR, 0.668, 95%CI, 0.530-0.843,
P=0.001), and the expression level of KCNMA1 was negatively correlated with
mitotic count (P=0.021). 3) Targeted sequencing of DNA samples showed that
point mutations at major KCNMA1 sites (p.N999S, p.N536H, p.D434G) were not
detected in recurrent meningiomas. However, the methylation levels of
CpG_133, CpG_711, CpG_1859 and CpG_2526 in KCNMA1 promoter in recurrent
meningiomas were higher than those in non-recurrent meningiomas, and the
difference was statistically significant (P < 0.05). Conclusions: Mitotic count ≥ 3/10 HPF, high WHO grade and maximum tumor
diameter ≥ 5 cm are prognostic factors for recurrence of meningiomas,
while down-regulation of KCNMA1 protein expression is an independent
prognostic factor for recurrence of meningiomas, and its molecular mechanism
may be related to the hypermethylation of KCNMA1 promoter. |