# Genetic Approaches to Optimize CAR T cells for Glioblastoma Therapy

> **NIH NIH R01** · NORTHWESTERN UNIVERSITY · 2020 · $527,915

## Abstract

Glioblastoma (GBM), the most frequently occurring and aggressive primary brain tumor, remains virtually
incurable. Thus, there is an urgent need to develop new therapies. Genetically modified T cells expressing
chimeric antigen receptors (CARs) have the potential to serve as a unique cytotoxic tool to specifically target
GBM. CAR T cell therapy has been successful for hematological malignancies, but multiple challenges posed
by the brain tumor environment require a multifaceted approach for CAR T cells to succeed for GBM. To study
this, we have developed a single-chain variable fragment (scFv) specific for IL13Rα2, a GBM-associated tumor
antigen, and have generated an IL13Rα2-CAR. IL13Rα2-CAR T cells only recognize IL13Rα2-positive glioma
cells and had anti-glioma activity in preclinical xenograft and immune-competent animal models. However,
tumors eventually recurred, paralleling the situation in humans. Major causes of treatment failure include (i) the
inability of CAR T cells to persist within an immunosuppressive tumor environment, (ii) antigen-loss variants
when a single antigen is targeted, and (iii) the inability of CAR T cells to efficiently traffic to tumor sites due to a
mismatch between chemokines produced by the tumor and chemokine receptors expressed by CAR T cells. In
mechanistic studies, we have demonstrated limited IL13Rα2-CAR T cell persistence and the development of
antigen-loss variants. In addition, we showed in xenograft models that transgenic expression of IL15 in CAR T
cells enhances their persistence and anti-glioma activity. However, these xenograft studies are limited; the goal
of this R01 is to perform mechanistic studies in immune-competent animal models and evaluate genetic
approaches to enhance the anti-glioma activity of IL13Rα2-CAR T cells. Thus, we now hypothesize that
IL13Rα2-CAR T cells can be further genetically engineered to optimize their anti-GBM activity by
enhancing their persistence, targeting multiple tumor antigens, and improving their trafficking to tumor
sites. Aim 1 investigates whether IL15-expressing CAR T cells can resist the immunosuppressive tumor
environment in syngeneic GBM models. Aim 2 optimizes CAR T cells to target both IL13Rα2 and EphA2, two
glioma-associated antigens. Aim 3 investigates if trafficking of CAR T cells to GBMs can be improved by the
transgenic expression of CCR2, a chemokine receptor that recognizes CCL2, a chemokine produced by GBMs.
At the conclusion of the grant, we will have addressed three major hurdles of CAR T cell therapy for GBM. While
we will use our data to justify the development of a future clinical study utilizing optimized IL13Rα2-CAR T cells
for patients with GBMs; our modified approach to T cell therapy should be applicable to a broad range of solid
tumors.

## Key facts

- **NIH application ID:** 9999697
- **Project number:** 5R01NS106379-03
- **Recipient organization:** NORTHWESTERN UNIVERSITY
- **Principal Investigator:** Irina V Balyasnikova
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $527,915
- **Award type:** 5
- **Project period:** 2018-09-30 → 2023-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9999697

## Citation

> US National Institutes of Health, RePORTER application 9999697, Genetic Approaches to Optimize CAR T cells for Glioblastoma Therapy (5R01NS106379-03). Retrieved via AI Analytics 2026-05-21 from https://api.ai-analytics.org/grant/nih/9999697. Licensed CC0.

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