Biology subjects

Binder, Z. A.

Publications and source records attributed to Binder, Z. A..

10 recordsLinked to original sources

bioRxiv2025

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

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. One Sentence SummaryDuring resection surgeries for brain tumors, delivering cell therapy at time of surgery could improve outcomes for patients.

cancer biology↗

bioRxiv2024

Hydralazine inhibits cysteamine dioxygenase to treat preeclampsia and senesce glioblastoma

The vasodilator hydralazine (HYZ) has been used clinically for [~] 70 years and remains on the World Health Organizations List of Essential Medicines as a therapy for preeclampsia. Despite its longstanding use and the concomitant progress toward a general understanding of vasodilation, the target and mechanism of HYZ have remained unknown. We show that HYZ selectively targets 2-aminoethanethiol dioxygenase (ADO) by chelating its metal cofactor and alkylating one of its ligands. This covalent inactivation slows entry of proteins into the Cys/N-degron pathway that ADO initiates. HYZs capacity to stabilize regulators of G-protein signaling (RGS4/5) normally marked for degradation by ADO explains its effect on blood vessel tension and comports with prior associations of insufficient RGS levels with human preeclampsia and analogous symptoms in mice. The established importance of ADO in glioblastoma led us to test HYZ in these cell types. Indeed, a single treatment induced senescence, suggesting a potential new HYZ-based therapy for this deadly brain cancer.

biochemistry↗

bioRxiv2024

Patient-derived glioblastoma organoids as real-time avatars for assessing responses to clinical CAR-T cell therapy

Patient-derived tumor organoids have been leveraged for disease modeling and preclinical studies, but rarely applied in real-time to aid with interpretation of patient treatment responses in clinics. We recently demonstrated early efficacy signals in a first-in-human, phase 1 study of dual-targeting chimeric antigen receptor T cells (EGFR-IL13R2 CAR-T cells) in patients with recurrent glioblastoma. Here we analyzed six sets of patient-derived glioblastoma organoids (GBOs) treated concurrently with the same autologous CAR-T cell products as patients in our phase 1 study. We found that CAR-T cell treatment led to target antigen reduction and cytolysis of tumor cells in GBOs, the degree of which correlated with CAR-T cell engraftment detected in patients cerebrospinal fluid (CSF). Furthermore, cytokine release patterns in GBOs mirrored patient CSF samples over time. Our findings highlight a unique trial design and GBOs as a valuable platform for real-time assessment of CAR-T cell bioactivity and insights into immunotherapy efficacy.

cancer biology↗

bioRxiv2024

Temporal multi-modal single-cell analyses reveal dynamic interactions of CAR-T cells with glioblastoma and targeting of antigen-negative neoplastic cells

CAR-T therapy is a promising new immunotherapy for cancers, but its efficacy for solid tumors requires improvement. A detailed understanding of the interplay between solid tumors and CAR-T cells is critical. Here we report temporal, multi-modal, single-cell profiling of patient-derived glioblastoma organoids with CAR-T treatment. We found that all tumor cell types responded to CAR-T cell activation and contributed to an initially anti-tumor, but subsequently pro-tumor and immune-inhibitory microenvironment, accompanied by CAR-T cell exhaustion. Unexpectedly, CAR-T treatment attenuated glioma stem-like states of both antigen-positive and antigen-negative neoplastic cells and reduced their proliferation via diffusible factors, including IFN{gamma}. Analysis of samples from additional patients, including those in clinical trials, supported these findings. Our study reveals the dynamic interplay among different tumor cells and T cells in adaptive responses to immunotherapy and identifies previously unappreciated benefits of CAR-T therapy directly on antigen-negative neoplastic cells that may be leveraged to enhance therapeutic efficacy.

cancer biology↗

bioRxiv2024

Brain-wide neuronal circuit connectome of human glioblastoma

Glioblastoma (GBM), a universally fatal brain cancer, infiltrates the brain and can be synaptically innervated by neurons, which drives tumor progression1-6. Synaptic inputs onto GBM cells identified so far are largely short-range and glutamatergic7-9. The extent of integration of GBM cells into brain-wide neuronal circuitry is not well understood. Here we applied a rabies virus-mediated retrograde monosynaptic tracing approach10-12 to systematically investigate circuit integration of human GBM organoids transplanted into adult mice. We found that GBM cells from multiple patients rapidly integrated into brain-wide neuronal circuits and exhibited diverse local and long-range connectivity. Beyond glutamatergic inputs, we identified a variety of neuromodulatory inputs across the brain, including cholinergic inputs from the basal forebrain. Acute acetylcholine stimulation induced sustained calcium oscillations and long-lasting transcriptional reprogramming of GBM cells into a more invasive state via the metabotropic CHRM3 receptor. CHRM3 downregulation suppressed GBM cell invasion, proliferation, and survival in vitro and in vivo. Together, these results reveal the capacity of human GBM cells to rapidly and robustly integrate into anatomically and molecularly diverse neuronal circuitry in the adult brain and support a model wherein rapid synapse formation onto GBM cells and transient activation of upstream neurons may lead to a long-lasting increase in fitness to promote tumor infiltration and progression.

cancer biology↗

bioRxiv2024

Armored Bicistronic CAR T Cells with Dominant-negative TGF-β Receptor II to Overcome Resistance in Glioblastoma.

Chimeric antigen receptor (CAR) T cells have shown significant efficacy in hematological diseases. However, CAR T therapy has demonstrated limited efficacy in solid tumors, including glioblastoma (GBM). One of the most important reasons is the immunosuppressive tumor microenvironment (TME), which promotes tumor growth and suppresses immune cells to eliminate tumor cells. The human transforming growth factor-beta (TGF-{beta}) plays a crucial role in forming the suppressive GBM TME and driving the suppression of the anti-GBM response. In order to mitigate TGF-{beta} mediated suppressive activity, we combined a dominant-negative TGF-{beta} receptor II (dnTGF{beta}RII) with our previous bicistronic CART-EGFR-IL13R2 construct, currently being evaluated in a clinical trial, to generate CART-EGFR-IL13R2-dnTGF{beta}RII, a tri-modular construct we are developing for clinical application. We hypothesized that this approach would more effectively subvert resistance mechanisms observed with GBM. Our data suggests that CART-EGFR-IL13R2-dnTGF{beta}RII significantly augmented T cell proliferation and enhanced functional responses, particularly in a TGF{beta}-rich tumor environment. Additionally, in vivo studies validated the safety and efficacy of the dnTGF{beta}RII cooperating with CARs in targeting and eradicating GBM in a NSG mouse model.

cancer biology↗

bioRxiv2022

The Radiogenomic and Spatiogenomic Landscapes of Glioblastoma, and Their Relationship to Oncogenic Drivers

Glioblastoma (GBM) is well-known for its molecular and spatial heterogeneity, which poses a challenge for precision therapies and clinical trial stratification. Here, in a comprehensive radiogenomics study of 358 GBMs, we investigated the associations between the imaging and spatial characteristics of the tumors with their cancer gene mutation status, as well as with the cross-sectionally inferred likely order of mutational events. We show that cross-validated machine learning analysis of multi-parametric MRI scans results in distinctive in vivo imaging signatures of several mutations, which are relatively more distinctive in homogeneous tumors which harbor only one of these mutations. These imaging signatures offer mechanistic insights into how various mutations influence the phenotype of the tumor and its surrounding infiltrated brain tissue via neovascularization and vascular leakage, increased cell density, invasion and migration, and other characteristics captured by respective imaging features. Furthermore, we found that spatial location and tumor distribution vary, depending on the GBMs molecular characteristics. Finally, distinct imaging and spatial characteristics were associated with cross-sectionally estimated evolutionary trajectories of the tumors. Collectively, our study establishes a panel of in vivo and clinically accessible imaging-AI biomarkers of GBM that reflect their molecular composition and oncogenic drivers.

bioengineering↗

bioRxiv2021

Locally secreted BiTEs complement CAR T cells by enhancing solid tumor killing

Bispecific T-cell engagers (BiTEs) are bispecific antibodies that redirect T cells to target antigen-expressing tumors. BiTEs can be secreted by T cells through genetic engineering and perform anti-tumor activity. We hypothesized that BiTE-secreting T cells could be a valuable T cell-directed therapy in solid tumors, with distinct properties in mono- or multi-valent strategies incorporating chimeric antigen receptor (CAR) T cells. Glioblastomas represent a good model for solid tumor heterogeneity and represent a significant therapeutic challenge. We detected expression of tumor-associated epidermal growth factor receptor (EGFR), EGFR variant III (EGFRvIII), and interleukin-13 receptor alpha 2 (IL13R2) on glioma tissues and glioma cancer stem cells. These antigens formed the basis of a multivalent approach, using a conformation-specific tumor-related EGFR targeting antibody (806) and Hu08, an IL13R2-targeting antibody, as the scFvs to generate new BiTE molecules. Compared with 806CAR T cells and Hu08CAR T cells, BiTE T cells demonstrated prominent activation, cytokine production, and cytotoxicity in response to target-positive gliomas. Superior response activity was also demonstrated in BiTE secreting bivalent targeting T cells compared with bivalent targeting CAR T cells, which significantly delayed tumor growth in a glioma mouse model. In summary, BiTEs secreted by mono- or multi- valent targeting T cells have potent anti-tumor activity in vitro and in vivo with significant sensitivity and specificity, demonstrating a promising strategy in solid tumor therapy.

cancer biology↗

bioRxiv2021

High Affinity Chimeric Antigen Receptor with Cross-Reactive scFv to Clinically Relevant EGFR Oncogenic Isoforms

Tumor heterogeneity is a key reason for therapeutic failure and tumor recurrence in glioblastoma (GBM). Our chimeric antigen receptor (CAR) T cell (2173 CAR T cells) clinical trial (NCT02209376) against Epidermal growth factor receptor (EGFR) variant III (EGFRvIII) demonstrated successful trafficking of T cells across the blood brain barrier into GBM active tumor sites. However, CAR T cell infiltration was associated only with a selective loss of EGFRvIII+ tumor, demonstrating little to no effect on EGFRvIII- tumor cells. Post-CAR T treated tumor specimens showed continued presence of EGFR amplification and oncogenic EGFR extracellular domain (ECD) missense mutations, despite loss of EGFRvIII. To address tumor escape, we generated an EGFR-specific CAR by fusing monoclonal antibody (mAb) 806 to a 4-1BB co-stimulatory domain. The resulting construct was compared to 2173 CAR T cells in GBM, using in vitro and in vivo models. 806 CAR T cells specifically lysed tumor cells and secreted cytokines in response to amplified EGFR, EGFRvIII, and EGFR-ECD mutations in U87MG cells, GBM neurosphere-derived cell lines, and patient-derived GBM organoids. 806 CAR T cells did not lyse fetal brain astrocytes or primary keratinocytes to a significant degree. They also exhibited superior antitumor activity in vivo when compared to 2173 CAR T cells. The broad specificity of 806 CAR T cells to EGFR alterations gives us the potential to target multiple clones within a tumor and reduce opportunities for tumor escape via antigen loss.

cancer biology↗

bioRxiv2019

The role of integrin αv and CD44 in GBM migration in human brain

Glioblastoma (GBM) is the most common primary adult malignant brain tumor. Recurrence is driven invading tumor cells that escape surgical resection and demonstrate resistance to standard-of-care chemotherapy and radiotherapy. A large body of research has been conducted on tumor cell motility. However, typical in vitro models make use of polystyrene culture dishes, which exhibit significantly different physical parameters than brain tissue. Here we report on the use of human organotypic brain slices as an ex vivo approach for the dynamic study of GBM cell motility. Temporal lobectomy tissue from epilepsy patients was obtained and cut into 350{micro}m thick slices. After the tissue slices had a weeks incubation for recovery, fluorescently labeled tumor cells were seeded. We then tracked individual tumor cells using time-lapse fluorescent confocal microscopy. Quantification of motility characteristics, including mean squared displacement, total path length, and consistency, allowed for comparison of different conditions, including knockdown of cell surface proteins integrin v (ITGAV) and CD44. Human organotypics demonstrated minimal variability across specimen in terms of motility parameters, including total path length, averaged instantaneous velocity, and consistency. Knockdown of the traditional motility protein ITGAV showed little effect on overall motility while knockdown of CD44 resulted in a significant reduction in both averaged instantaneous velocity and total path length. When the same parameters were examined using Matrigel, ITGAV and CD44 both showed decreased motility, highlighting the impact of the physical environment on cell behavior. Finally, cell motility in mouse organotypic slices was decreased when compared to human organotypic slices. Here we demonstrate the use of human organotypic brain slices in the study of GBM cell invasion. This model system offers a physiologically-relevant environment in which to examine the dynamic process of cell motility.

cancer biology↗