SynNotch-CAR T cells overcome challenges of specificity, heterogeneity, and persistence in treating glioblastoma

Abstract

Treatment of solid cancers with chimeric antigen receptor (CAR) T cells is plagued by the lack of ideal target antigens that are both absolutely tumor specific and homogeneously expressed. We show that multi-antigen prime-and-kill recognition circuits provide flexibility and precision to overcome these challenges in the context of glioblastoma. A synNotch receptor that recognizes a specific priming antigen, such as the heterogeneous but tumor-specific glioblastoma neoantigen epidermal growth factor receptor splice variant III (EGFRvIII) or the central nervous system (CNS) tissue-specific antigen myelin oligodendrocyte glycoprotein (MOG), can be used to locally induce expression of a CAR. This enables thorough but controlled tumor cell killing by targeting antigens that are homogeneous but not absolutely tumor specific. Moreover, synNotch-regulated CAR expression averts tonic signaling and exhaustion, maintaining a higher fraction of the T cells in a naïve/stem cell memory state. In immunodeficient mice bearing intracerebral patient-derived xenografts (PDXs) with heterogeneous expression of EGFRvIII, a single intravenous infusion of EGFRvIII synNotch-CAR T cells demonstrated higher antitumor efficacy and T cell durability than conventional constitutively expressed CAR T cells, without off-tumor killing. T cells transduced with a synNotch-CAR circuit primed by the CNS-specific antigen MOG also exhibited precise and potent control of intracerebral PDX without evidence of priming outside of the brain. In summary, by using circuits that integrate recognition of multiple imperfect but complementary antigens, we improve the specificity, completeness, and persistence of T cells directed against glioblastoma, providing a general recognition strategy applicable to other solid tumors.

Fibroblast Activation Protein (FAP)-Targeted CAR-T Cells: Launching an Attack on Tumor Stroma

“…The synNotch-CAR targets multiple antigens in the TME and utilizes prime-and-kill circuits to achieve specific and controlled killing of tumor cells. 30 Tumor-associated antigens (TAAs) are overexpressed by tumor cells and, at low levels, by healthy cells. The three currently approved CAR-T cell therapies, Yescarta, Kymriah and Tecartus, target CD19 on B lymphocytes.…”

Advances in Immunotherapy for Adult Glioblastoma

“…An in-depth review of bispecific antibodies, including BiTEs, was recently presented by Lim et al [ 117 ]. Moreover, investigators recently developed multi-antigen prime-and-kill synNotch-CAR T cells that use a dual receptor circuit, the first of which detects EGFRvIII or a brain-specific myelin oligodendrocyte glycoprotein to induce expression of CARs against EphA2 and IL13Rα2 [ 118 ]. In comparison to constitutively active anti-EGFRvIII/EphA2/IL13Rα2 CAR T cells, synNotch-CAR T cells showed greater anti-tumor efficacy without off-tumor toxicity.…”

Chimeric Antigen Receptor-T Cells: A Pharmaceutical Scope

“…SynNotch receptors recognize one specific tumor antigen and then transcriptional activation domains are released, promoting the local expression of a CAR ( Han et al, 2019 ). Moreover, synNotch-regulated CAR expression prevents constitutive signaling and exhaustion, maintaining a higher fraction of the T cells in a naïve/stem cell memory state ( Choe et al, 2021 ). Synthetic Notch (synNotch) receptors have been applied in CAR-T cells to improve safety.…”