# Development of novel synNotch CART cell therapy in adult patients with recurrent EGFRvIII+ glioblastoma

> **NIH NIH U19** · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · 2022 · $253,654

## Abstract

Development of safe and effective (CAR)-transduced T cell (CART) therapy for glioblastoma (GBM) needs to
overcome multiple challenges, including on-target off-tumor toxicity, heterogeneity of antigen expression, and
exhaustion of CART cells. There are no GBM-specific surface antigens that are uniformly present in tumor
tissue. Mutant epidermal growth factor receptor (EGFRvIII), for example, is GBM-specific but its expression is
heterogenous within the tumor. On the other hand, while non-mutant GBM-associated antigens (GAAs), such
as EphA2 and IL-13Rα2, are more uniformly expressed in GBM, their expression in other organs outside the
central nervous system (CNS) raises concern for off-tumor toxicity. As a way to safely and effectively target
GAAs, we have adopted a novel synthetic Notch “synNotch” receptor system and developed innovative T cells.
In this system, the first antigen, which is expressed exclusively on brain or GBM cells (e.g. EGFRvIII), primes
the T cells to induce expression of a CAR that recognizes IL-13Rα2 and EphA2, thereby eradicating GBM cells
expressing either EphA2 or IL-13α2. Our data show that EGFRvIII-synNotch primed IL-13Rα2/EphA2 CAR are
effectively but restrictedly activated by EGFRvIII as the GBM-specific signal, leading to complete eradication of
patient-derived xenografts with heterogeneous EGFRvIII expression but without attacking IL-13Rα2/EphA2-
positive cells outside of CNS. Furthermore, these synNotch-CAR T cells were significantly more efficacious
than conventional, constitutively expressed IL-13Rα2/EphA2 CAR T cells, and were associated with excellent
persistence (>100 days in vivo). Taken together, our data indicate that synNotch CAR T cells can revolutionize
the CAR T therapy for solid cancers by overcoming the off-tumor toxicity, antigen heterogeneity and lack of
persistence. Our goals are to establish the process for development of Good Manufacturing Practice
products of EGFRvIII-primed IL-13Rα2/EphA2 CAR T cells, to obtain an Investigational New Drug (IND)
approval, and to develop and conduct a phase I trial in patients with recurrent GBM. In Aim 1, we will
generate GMP-grade lentiviral vector and establish standard procedures for GMP-grade manufacturing of the
EGFRvIII-primed IL-13Rα2/ EphA2 CAR T cells. In Aim 2, we will complete both in vitro and in vivo studies that
are required for submission of an IND. In vitro studies include confirmation of antigen-specific priming, specific
cytotoxicity, and absence of replication competent lentivirus. In vivo, we will confirm preclinical efficacy,
toxicology, tissue biodistribution, and engraftment as well as persistence of synNotch CART cells. In Aim 3, in
a first-in-human phase 1 clinical trial, we will determine safety and toxicity of the synNotch CART cells in
patients with recurrent EGFRvIII+ GBM. In a subsequent expansion cohort, in patients with recurrent EGFRvIII+
GBM, we will infuse synNotch CART cells intravenously prior to surgery, and then evaluate inf...

## Key facts

- **NIH application ID:** 10487528
- **Project number:** 5U19CA264338-02
- **Recipient organization:** UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
- **Principal Investigator:** Hideho Okada
- **Activity code:** U19 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $253,654
- **Award type:** 5
- **Project period:** 2021-09-10 → 2026-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10487528

## Citation

> US National Institutes of Health, RePORTER application 10487528, Development of novel synNotch CART cell therapy in adult patients with recurrent EGFRvIII+ glioblastoma (5U19CA264338-02). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10487528. Licensed CC0.

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