bims-malgli 2025-03-02 papers

Issue of 2025–03–02
five papers selected by
Oltea Sampetrean, Keio University

Proc Natl Acad Sci U S A. 2025 Mar 04. 122(9): e2415557122

Radiation-induced cellular plasticity primes glioblastoma for forskolin-mediated differentiation.

Ling He, Daria Azizad, Kruttika Bhat, Angeliki Ioannidis, Carter J Hoffmann, Evelyn Arambula, Mansoureh Eghbali, Aparna Bhaduri, Harley I Kornblum, Frank Pajonk.

Glioblastoma (GBM) is the deadliest brain cancer in adults, and all patients succumb to the tumor. While surgery followed by chemoradiotherapy delays disease progression, these treatments do not lead to tumor control, and targeted therapies or biologics have failed to further improve survival. Utilizing a transient radiation-induced state of multipotency, we used the adenylcyclase activator forskolin to alter the fate of irradiated glioma cells. The effects of the combined treatment on neuronal marker expression, cell cycle distribution, and proliferation were studied. Gene expression profiling was conducted using bulk RNA-seq. Changes in cell populations were investigated using single-cell RNA-seq. Effects on glioma stem cells (GSCs) were studied in extreme limiting dilution assays, and the effects on median survival were studied in both syngeneic and PDOX mouse models of GBM. The combined treatment induced the expression of neuronal markers in glioma cells, reduced proliferation, and led to a distinct gene expression profile. scRNA-seq revealed that the combined treatment forced glioma cells into a microglia- and neuron-like phenotype. In vivo, this treatment led to a loss of GSCs and prolonged median survival. Collectively, our data suggest that revisiting a differentiation therapy with forskolin in combination with radiation could lead to clinical benefit.

Keywords: glioblastoma; glioma stem cells; radiotherapy

DOI: https://doi.org/10.1073/pnas.2415557122

Neuro Oncol. 2025 Feb 25. pii: noaf055. [Epub ahead of print]

Multiplexed epigenetic memory editing using CRISPRoff sensitizes glioblastoma to chemotherapy.

Katie Lin, Christopher Zou, Akane Hubbard, Sasha Sengelmann, Laine Goudy, I-Ching Wang, Rohit Sharma, Joanna Pak, Kyla Foster, Tomoko Ozawa, John F de Groot, Joanna Phillips, Harish N Vasudevan, David R Raleigh, Alexander Marson, Niren Murthy, Luke A Gilbert, Mitchel S Berger, S John Liu.

BACKGROUND: Glioblastoma (GBM) carries a poor prognosis, and new therapeutic strategies are necessary to improve outcomes for patients with this disease. Alkylating chemotherapies including temozolomide (TMZ) and lomustine (CCNU) are critical for treating GBM, but resistance mechanisms, including hypomethylation of O6-methylguanine-DNA methyltransferase (MGMT) promoter, undermine treatment. CRISPRoff is a programmable epigenetic memory editor that can induce stable and heritable gene silencing after transient delivery, and we hypothesize that CRISPRoff could potentiate the activity of TMZ and CCNU through long term suppression of target genes.
METHODS: We transiently delivered CRISPRoff mRNA along with sgRNAs against target genes using both electroporation and lipid nanoparticles (LNPs) into established GBM cell lines, patient-derived primary GBM cultures, and orthotopic GBM xenografts. Gene repression, specificity, and stability was measured by RT-qPCR, Western blot, bisulfite sequencing, and RNA-sequencing. Sensitivity to chemotherapies was measured by cell viability dose response, microscopy, and bioluminescence imaging. Genome-wide mapping of CCNU sensitizers was performed using CRISPRi screens.
RESULTS: CRISPRoff induced complete suppression of MGMT and sensitization to TMZ that were stable for over 8 months of continuous cell propagation. GBM orthotopic tumors treated with CRISPRoff against MGMT demonstrated sensitivity to TMZ in vivo, and CRISPRoff delivery resulted in chemosensitivity in patient-derived primary GBM. Genome-wide CRISPRi screens identified combinatorial genetic vulnerabilities (BRIP1, FANCE) that were targetable by multiplexed CRISPRoff to achieve sensitization to CCNU.
CONCLUSION: Transient delivery of a site-specific epigenetic memory can induce stable, complete, and multiplexed suppression of target genes for therapeutic application in GBM.

Keywords: CCNU; CRISPR; epigenetic editing; glioblastoma; temozolomide

DOI: https://doi.org/10.1093/neuonc/noaf055

bioRxiv. 2025 Feb 15. pii: 2025.02.11.637648. [Epub ahead of print]

CAR-T cells locally delivered in porcine decellularized matrix hydrogels enhance survival in post-resection glioblastoma.

Meghan Logun, Sabrina L Begley, Kelly Hicks, Jungmin Park, Logan Zhang, Zev A Binder, Donald M O'Rourke.

Patients diagnosed with glioblastoma (GBM) receive a devastating prognosis of less than 15 months, and recurrence of GBM is most often local, suggesting that regional therapies would serve both immediate and long-term needs of patients. Here, we investigate a biomaterials-based approach for local delivery of chimeric antigen receptor (CAR) T cells using a murine model of partial GBM resection that mimics patient recurrence. We demonstrate that hydrogel delivery of CAR T cells directly into the intracranial resection cavity can stably implant cellular immunotherapies against CNS solid tumors, and significantly prolongs survival in recurrent GBM-bearing mice compared to those receiving resection alone.

DOI: https://doi.org/10.1101/2025.02.11.637648

Neuro Oncol. 2025 Feb 25. pii: noaf056. [Epub ahead of print]

Synapsin III promotes neuronal-like transdifferentiation of glioblastoma stem cells by disrupting JAG1-Notch1 interaction.

Yilin Deng, Zheng Yuan, Xiong Jin, Zekun Wang, Rui Gong, Shuai Ren, Jong Bae Park, Bingyang Shi, Jinlong Yin.

BACKGROUND: Glioblastoma (GBM), a formidable and highly aggressive form of brain cancer, is predominantly driven by GBM stem cells (GSCs), which are characterized by their ability for self-renewal and aberrant differentiation. Targeting the terminal differentiation of GSCs represents a promising therapeutic strategy. This study aimed to elucidate the role of synapsin III (SYN3) in driving the differentiation of GSCs into neuron-like cells and its effect on the tumor-suppressive pathways in GBM.
METHOD: Proliferation assays, limited dilution assays, immunocytochemistry, western blot, RT-qPCR, and GSC tumor models were used to determine gene function and assess the role of γ-secretase inhibitors. Co-immunoprecipitation and microscale thermophoresis were conducted to explore the underlying regulatory mechanisms. Intracranial orthotopic injection of adeno-associated virus (AAV) was performed to evaluate therapeutic potential.
RESULTS: We demonstrate that SYN3, uniquely within the synapsin family, acts as a tumor suppressor by steering GSCs toward neuronal-like transdifferentiation. Mechanistically, SYN3 enhances the expression of Neuregulin 3 (NRG3), which serves as a non-canonical antagonist of Notch signaling by competitively binding to specific epitopes within the EGF-like domain of JAG1, a critical site for the canonical engagement of Notch receptors. This critical interaction disrupts the JAG1-Notch1 signaling pathway, a key mechanism driving GSCs toward neuronal-like transdifferentiation, thereby reducing their stemness. Furthermore, SYN3 demonstrated significant antineoplastic activity in a mouse model harboring GSCs. AAV-mediated overexpression of SYN3 markedly impeded GBM progression.
CONCLUSION: Our research reveals the therapeutic potential of SYN3 in regulating GSC fate and offers a novel differentiation-based approach for GBM therapy.

Keywords: Glioblastoma; Glioblastoma stem cells; Neuronal-like transdifferentiation; Notch signaling; Synapsin III

DOI: https://doi.org/10.1093/neuonc/noaf056

Nature. 2025 Feb 26.

Programs, origins and immunomodulatory functions of myeloid cells in glioma.

Tyler E Miller, Chadi A El Farran, Charles P Couturier, Zeyu Chen, Joshua P D'Antonio, Julia Verga, Martin A Villanueva, L Nicolas Gonzalez Castro, Yuzhou Evelyn Tong, Tariq Al Saadi, Andrew N Chiocca, Yuanyuan Zhang, David S Fischer, Dieter Henrik Heiland, Jennifer L Guerriero, Kevin Petrecca, Mario L Suva, Alex K Shalek, Bradley E Bernstein.

Gliomas are incurable malignancies notable for having an immunosuppressive microenvironment with abundant myeloid cells, the immunomodulatory phenotypes of which remain poorly defined1. Here we systematically investigate these phenotypes by integrating single-cell RNA sequencing, chromatin accessibility, spatial transcriptomics and glioma organoid explant systems. We discovered four immunomodulatory expression programs: microglial inflammatory and scavenger immunosuppressive programs, which are both unique to primary brain tumours, and systemic inflammatory and complement immunosuppressive programs, which are also expressed by non-brain tumours. The programs are not contingent on myeloid cell type, developmental origin or tumour mutational state, but instead are driven by microenvironmental cues, including tumour hypoxia, interleukin-1β, TGFβ and standard-of-care dexamethasone treatment. Their relative expression can predict immunotherapy response and overall survival. By associating the respective programs with mediating genomic elements, transcription factors and signalling pathways, we uncover strategies for manipulating myeloid-cell phenotypes. Our study provides a framework to understand immunomodulation by myeloid cells in glioma and a foundation for the development of more-effective immunotherapies.

DOI: https://doi.org/10.1038/s41586-025-08633-8