A human tumor-immune organoid model of glioblastoma

Cell Reports · 2026-01-01 · 8 citations

A major obstacle to identifying effective therapies for the aggressive brain tumor glioblastoma is the lack of human-specific, immunocompetent models that reflect the human tumor microenvironment. To address this, we developed the immune-human organoid tumor transplantation (iHOTT) model, an autologous co-culture platform that integrates patient-derived tumor cells and matched peripheral blood mononuclear cells within human cortical organoids to enable the study of patient-specific immune responses and tumor-immune interactions. This platform preserves tumor and immune populations, immune signaling, and cell-cell interactions observed in patient tumors. Treatment of iHOTT with pembrolizumab, a checkpoint inhibitor, mirrors cell-type shifts and cell-cell interactions observed in patients. T cell receptor (TCR) sequencing further reveals pembrolizumab-driven expansion of stem-like CD4 T cell clonotypes exhibiting patient-specific repertoires. These findings establish iHOTT as a physiologically relevant platform for exploring autologous tumor-immune interactions and underscore the need for antigen-targeted strategies to enhance immunotherapy in glioblastoma.

Vaccine therapy for pediatric high-grade glioma: current landscape, challenges, and future directions

Journal of Neuro-Oncology · 2026-01-01

Pediatric high-grade gliomas (pHGG) are among the most aggressive childhood brain tumors, with limited treatment options and poor prognosis. Vaccine-based immunotherapy offers a promising strategy by leveraging tumor-specific or associated antigens to stimulate durable anti-tumor immune responses with minimal toxicity. This review outlines the scientific rationale for vaccine therapies in pHGG, detailing key targets such as glioma-associated antigens (EphA2, IL-13Rα2, survivin), driver mutation–derived neoantigens (H3.3K27M, TP53, IDH1), and viral antigens (CMV pp65). We evaluate current vaccine platforms, including peptide vaccines, dendritic cell vaccines, mRNA-based vaccines, and neoantigen-personalized approaches, highlighting early-phase clinical trial results that demonstrate safety and immunogenicity. Despite encouraging preliminary data, several challenges hinder clinical translation, including the distinct immune environment in the central nervous system, intratumoral heterogeneity, low mutational burden, immunosuppressive microenvironments, steroid use, and logistical hurdles in vaccine production and trial design. Future research must address these barriers through optimized antigen selection, combinatorial therapies, novel delivery systems, and pediatric-specific immune profiling. With continued multidisciplinary collaboration, vaccine therapies may emerge as a meaningful addition to the therapeutic arsenal for children with pHGG.

Immunotherapeutic targeting of NY-ESO-1 in malignant meningiomas with TCR-transduced T-cells

Journal of Neuro-Oncology · 2025-08-13 · 1 citations

EXTH-02. GliomaPDOX – A direct brain-to-brain glioma xenograft library for drug discovery and development

Neuro-Oncology · 2025-11-01

Abstract Cancer drug discovery and development rely on preclinical models that accurately reflect the molecular and functional characteristics of human tumors, while accounting for in vivo factors that influence drug efficacy, such as pharmacokinetics, metabolism and toxicity. Malignant gliomas are highly aggressive brain tumors that develop within the brain parenchyma, where their heterogeneous cellular composition engage in complex interactions with highly specialized brain cells, and a blood brain barrier that restricts drug penetration. When removed from this native environment, such as in culture or heterotopic in vivo environments (e.g., flank), gliomas either lose their molecular diversity or fail to grow altogether. Therefore, there is a critical need for physiologically relevant models that capture both the intra and inter-tumor diversity of glioma, as well as the organismal context required for drug development. Here, we present GliomaPDOX – a direct brain to brain glioma orthotopic xenograft biobank, consisting of more than 200 unique models that faithfully recapitulate the key molecular, histopathological, and proliferative features of their parental tumors. By incorporating a non-invasive, secreted reporter system to monitor tumor burden in real time—including drug-induced changes in intracranial tumor growth—we demonstrate the utility of GliomaPDOX for therapeutic evaluation. Together, this robust platform provides a physiologically relevant system to accelerate drug discovery and development for glioma.

A Human Tumor-Immune Organoid Model of Glioblastoma

bioRxiv (Cold Spring Harbor Laboratory) · 2025-06-20 · 3 citations

A major obstacle to identifying effective therapies for the aggressive brain tumor glioblastoma is the lack of human-specific, immunocompetent models that reflect the human tumor microenvironment. To address this, we developed the immune-Human Organoid Tumor Transplantation (iHOTT) model. This is an autologous co-culture platform that integrates patient-derived tumor cells and matched peripheral blood mononuclear cells (PBMCs) within human cortical organoids, enabling the study of the patient-specific immune response to the tumor and tumor-immune interactions. This platform preserves tumor and immune populations, immune signaling, and cell-cell interactions observed in patient tumors. Treatment of iHOTT with pembrolizumab, a checkpoint inhibitor, mirrored cell type shifts and cell interactions observed in patients. TCR sequencing further revealed pembrolizumab-driven expansion of stem-like CD4-T-cell clonotypes exhibiting patient-specific repertoires. These findings establish iHOTT as a physiologically relevant platform for exploring autologous tumor-immune interactions and underscore the critical need for antigen-targeted strategies to enhance immunotherapy in glioblastoma.

IMMU-36. iHOTT: A Human Tumor–Immune Organoid Model for Interrogating Immunotherapy Response in Glioblastoma

Neuro-Oncology · 2025-11-01

Abstract Glioblastoma is a highly aggressive brain tumor with a dismal prognosis, in part due to its ability to evade immune detection through an immunosuppressive microenvironment. While immune checkpoint inhibitors have shown promise in other cancers, glioblastoma remains largely resistant to immunotherapy. There is a critical need for physiologically relevant, patient-derived human models to investigate mechanisms of immune resistance and response. Here, we developed and validated iHOTT, a human tumor–immune organoid transplantation system, in which patient glioblastoma cells and matched circulating immune cells (PBMCs) are co-cultured within mature human cortical organoids. This model was evaluated using single-cell RNA sequencing, cytokine profiling, immunofluorescence, and T cell receptor (TCR) sequencing, and was benchmarked against pembrolizumab-treated and untreated patient tumor samples. iHOTT preserved diverse tumor and immune compartments, supported cytokine production in the presence of both tumor and immune cells, and demonstrated robust immune activation at both the transcriptional and protein levels. Upon pembrolizumab treatment, the model mirrored multiple aspects of patient immune responses, including expansion of T cells, B cells, and ILCs. Cell–cell interaction analyses showed increased signaling involving unconventional T cell subsets such as MAIT and γδ T cells, which were mirrored between patient samples and iHOTT. Furthermore, we noted upregulation of immunologically relevant pathways, including complement, CD226, and CD70 in both iHOTT and patients. TCR sequencing revealed increased clonal diversity driven by CD4 T cells in treated iHOTT samples, consistent with patient tumor profiles. Clonotype clustering revealed largely private, patient-specific responses in both iHOTT and patient datasets, reflecting the inter-tumoral heterogeneity that may underlie inconsistent responses to PD-1 blockade in glioblastoma. This autologous human organoid platform provides a scalable, multimodal system to model patient-specific tumor–immune dynamics, dissect immunotherapy mechanisms, and support biomarker discovery and therapeutic development in glioblastoma.

PATH-82. Multivariate survival risk stratification in GBMs lacking necrosis and microvascular proliferation

Neuro-Oncology · 2025-11-01

Abstract IDH-wildtype diffuse astrocytic gliomas with lower-grade histological features alongside specific genomic alterations (e.g., TERT promoter mutation, EGFR amplification, gain Chr7/loss Chr10) are presently diagnosed as glioblastoma, IDH-wildtype (GBM) under WHO 2021 criteria. Previous analyses categorized these into “early/evolving” or “surgically undersampled” GBMs. However, it remains unclear whether these tumors share identical clinical and prognostic features with histologic GBM. Data from IDH-wt molecular GBMs across six institutions were aggregated. Astrocytic IDH-wt tumors of any grade that lacked necrosis or microvascular proliferation on initial histopathology yet exhibited molecular alterations consistent with WHO 2021 GBM diagnosis, were included. Recursive partitioning analysis (RPA) was used to categorize patients into risk groups based on overall survival (OS). Among the 264 patients included, the median age was 60 years (IQR: 51-67), with 162 (61%) being male. Additionally, 210 (80%) were diagnosed since 2016, and 216 (82%) received chemoradiation plus adjuvant temozolomide (GBM-SOC). The median follow-up period was 3.9 years (95% CI: 3.4, 4.6). Identified univariate risk factors for worse survival included older age, biopsy-only status, EGFR amplification, absence of initial adjuvant treatment, low post-operative KPS, and tumor histologic grade of 3. RPA divided patients into four distinct survival groups (p<0.0001). The “best” risk group comprised lower grade (<=2) tumors (n=58, median OS (mOS): 40 months (95% CI: 33, NA)). The “second best” group included Grade 3 patients receiving GBM-SOC without steroids during initial RT (n=139, mOS: 25 months (95% CI: 22, 28)). The “second worst” group encompassed Grade 3 patients who were on steroids during initial RT while receiving GBM-SOC (n=38, mOS: 15 months (95% CI: 13, 20)). The “worst” group contained Grade 3 tumors that had not undergone SOC (n=29, mOS: 9 months (95% CI: 4, 26)). These data show that histologic features and treatment are strong indicators of patient outcomes.

Characterizing Intra- and Peritumoral Connectivity in Human Gliomas using Pseudo-Resting State Functional MRI Derived from DSC Perfusion MRI

Proceedings on CD-ROM - International Society for Magnetic Resonance in Medicine. Scientific Meeting and Exhibition/Proceedings of the International Society for Magnetic Resonance in Medicine, Scientific Meeting and Exhibition · 2025-09-16

Motivation: Clinical use of resting-state fMRI (rs-fMRI) to study brain tumor biology is not widespread, due in part by time and cost constraints. Goal(s): Finding an alternative to traditional rs-fMRI for studying brain tumor biology and functional connectivity. Approach: Deriving "pseudo" rs-fMRI from routine clinical DSC perfusion MRI, and using intra-tumoral connectivity (ITC) and dynamic time warping (DTW) distance to characterize tumor infiltration and heterogeneity. Results: "Pseudo" rs-fMRI derived from clinical DSC perfusion MRI may be useful for monitoring brain tumor patients. ITC and peritumoral connectivity patterns are unique to different glioma molecular subtypes, suggesting differences in the underlying biology. Impact: "Pseudo" rs-fMRI derived from clinical DSC perfusion MRI may be useful for monitoring brain tumor patients. Tumor infiltration and heterogeneity associated with different glioma molecular subtypes can be revealed by connectivity patterns within the tumor and with adjacent, normal-appearing tissue.

Advanced imaging characterization of post-chemoradiation glioblastoma stratified by diffusion MRI phenotypes known to predict favorable anti-VEGF response

Journal of Neuro-Oncology · 2025-04-14

Abstract Purpose Recurrent glioblastomas showing a survival benefit from anti-VEGF agents are known to exhibit a distinct diffusion MRI phenotype. We aim to characterize advanced imaging features of this glioblastoma subset. Methods MRI scans from 87 patients with IDH-wildtype glioblastoma were analyzed. All patients had completed standard chemoradiation and were anti-VEGF-naïve. Contrast-enhancing tumor segmentations were used to extract: the lowest peak of the double gaussian distribution of apparent diffusion coefficient values (ADC L ) calculated from diffusion MRI, relative cerebral blood flow (rCBV) values from perfusion MRI, MTR asym @ 3ppm from pH-weighted amine CEST MRI, quantitative T 2 and T 2 * relaxation times (qT 2 and qT 2 *), T 1 w subtraction map values, and contrast-enhancing tumor volume. Lesions were categorized as high- or low-ADC L using a cutoff of 1240 µm 2 /s, according to previous studies. Results High-ADC L lesions showed significantly lower rCBV (1.02 vs. 1.28, p = 0.0057), higher MTR asym @ 3ppm (2.36% vs. 2.10%, p = 0.0043), and higher qT 2 (114.8 ms vs. 100.9 ms, p = 0.0094), compared to low-ADC L lesions. No group differences were seen in contrast-enhancing tumor volume, T 1 w subtraction map values, and qT 2 *, nor in clinical variables such as sex category, MGMT status, and EGFR status. Finally, no clear group-specific preferential locations were seen. Conclusion Post-chemoradiation glioblastomas with a diffusion MRI phenotype that is known to predict a favorable response to anti-VEGF (ADC L ≥1240 µm 2 /s) have distinct biological features, with different perfusion and metabolic characteristics, and T 2 relaxation times.

A single-institution retrospective study of multicentric gliomas stratified by <i>IDH</i> mutational status.

Journal of Clinical Oncology · 2025-05-28

2078 Background: Multicentric glioma (MCG) is a subset of diffuse glioma that can be synchronous or metachronous and is defined as the occurrence of two or more tumor foci, with separation of FLAIR (Fluid-attenuated Inversion Recovery) hyperintensity on MRI. MCG has not been extensively studied in studies stratifying IDH wild-type and mutant gliomas. This large single-institution study investigates the prevalence of MCG, examines the prognostic implications of MCG, and characterizes metachronous MCG (mMCG) in a cohort that has been stratified by IDH mutational status. Methods: In this IRB approved UCLA study, we identified diffuse glioma patients with known IDH mutational status and adequate MRI studies. Patients with multiple lesions on the MRI study pre-surgery or up to 3 months post-surgery were considered synchronous MCG (sMCG). Patients who developed a new independent lesion at least 6 months after initial surgery were considered mMCG. To qualify as MCG, we identified additional tumors on MRI that had no overlapping FLAIR borders and met one or more of the following: pathologically confirmed with biopsy, exhibited growth and thickening over time, and developed or increased in enhancement. Difference in prevalence was compared using Student’s t-test. Kaplan-Meier and Cox-multivariate analyses were used to analyze OS and time to metachronous (TtM) appearance. Results: We identified 911 consecutive IDH wild-type, high-grade diffuse glioma patients from 2013-2023 and 515 consecutive IDH mutant patients from 2007-2024 with pre-surgical MRI or MRI within three months of initial surgery. From the examined cohort, we found 39 IDH mutants with 21 sMCG and 18 mMCG and 153 IDH wild types with 95 sMCG and 63 mMCG. In eight IDH wild-type cases but no IDH mutant cases, mMCG arose from sMCG patients. We found that MCG had higher prevalence in IDH wild-types than in mutants (WT = 16%, Mut = 7%, p &lt; 0.0001), and IDH mutant MCG showed more male predominance than IDH wild-type MCG (Mut = 73%, WT = 58%, p &lt; 0.0001). In IDH mutant patients, mMCG, but not sMCG, was associated with lower OS (mMCG: HR = 2.476, p = 0.0115; sMCG: HR = 0.6437, p = 0.5027). However, in IDH wild types both sMCG and mMCG and were associated with lower OS (mMCG: HR = 1.589, p = 0.0025; sMCG: HR = 1.347, p = 0.0332). There was no difference in TtM between the two groups (HR = 0.5738, p = 0.5318). Amongst patients with multiple biopsied lesions, IDH wild types had consistent pathologies between lesions in all examined patients (29/29), but 71% of IDH mutants exhibited different pathologies between lesions (5/7). Conclusions: Our study examined a cohort of adult diffuse gliomas stratified by IDH mutational status and shows MCG is less common in IDH mutant gliomas and sMCG is not associated with worse prognosis. Further studies to identify molecular features underlying MCG will be valuable. Notes: For mMCG, FLAIR overlap might have occurred had the new lesion been observed synchronously.