277 SynNotch-CAR T cells demonstrate potent anti-tumor efficacy in a preclinical immunocompetent mouse model for glioblastoma
Bibliographic record
Abstract
<h3>Background</h3> Glioblastoma multiforme (GBM) is the most common primary malignant brain tumor. Recent immunotherapy studies, including CAR T cells, have shown limited success in GBM due to multiple barriers, such as on-target off-tumor toxicity, the blood-brain barrier, and the immunosuppressive tumor microenvironment (TME). To overcome these challenges, we recently adopted a novel synthetic Notch ‘synNotch’ receptor system and developed innovative T cell (‘synNotch-CART’) circuits based on the ‘prime-and-kill’ strategy.<sup>1</sup> Here, we investigated the GBM-homing and efficacy of SynNotch-CAR T cells using a preclinical immunocompetent mouse model, which, similar to GBM patients, is poorly immunogenic and unresponsive to immune checkpoint blockade.<sup>2</sup> <h3>Methods</h3> Epidermal growth factor receptor (EGFR) amplification and overexpression occur in approximately 40–60% of GBM patients.<sup>3</sup> Therefore, we first expressed the extracellular domain of mouse EGFR in a syngeneic glioma cell line, SB28 (SB28-mEGFR). Next, we developed a novel synNotch-CAR (mBSYNC) circuit in which the brain-specific antigen, brevican (BCAN) primes the T cells to induce the expression of a CAR that recognizes and kills EGFR-positive cells (mBSYNC synNotch-CAR T cells). 8-week-old C57BL/6 mice bearing the SB28-mEGFR cells in the frontal lobe received a single intravenous infusion of mBSYNC synNotch-CAR or conventional CAR T cells with constitutive CAR expression on day 8 following the tumor inoculation. Trafficking of CAR T cells and host immune cells into the brains was evaluated using high-dimensional flow cytometry. <h3>Results</h3> Constitutive a-EGFR CAR and mBSYNC synNotch-CAR T cells mediated antigen-specific cytotoxicity against SB28-mEGFR cells <i>in vitro</i>. In an <i>in vivo</i> prospective study, on day 12 after T cell infusion, mBSYNC synNotch-CAR T cells demonstrated a higher persistence, increased infiltration of endogenous CD86<sup>+</sup> and MHC-II<sup>hi</sup> macrophages and decreased infiltration of CD206<sup>+</sup> macrophages into the GBM TME compared to either untransduced or constitutive a-EGFR CAR T-cells. Lastly<i>,</i> a single intravenous infusion of mBSYNC synNotch-CAR T cells significantly (p < 0.01) prolonged the survival and completely eradicated the aggressive tumor in 3 of 10 mice bearing SB28-mEGFR gliomas. Conclusions mBSYNC synNotch-CAR T cells, but not conventional CAR T cells, effectively home into the brain TME, persist, and eliminate the tumor in an immunocompetent setting. The induced pro-inflammatory activity of endogenous myeloid cells could contribute to the anti-tumor efficacy mediated by synNotch-CAR T cells. Ongoing studies are aimed at the thorough characterization of tumor cells, host immune cells, and synNotch-CAR T cells in the brain TME. <h3>References</h3> Choe JH, Watchmaker PB, Simic MS, Gilbert RD, Li AW, Krasnow NA, Downey KM, Yu W, Carrera DA, Celli A, <i>et al.</i> SynNotch-CAR T cells overcome challenges of specificity, heterogeneity, and persistence in treating glioblastoma. <i>Sci Transl Med</i> 2021;<i>13</i>. 10.1126/scitranslmed.abe7378. Genoud V, Marinari E, Nikolaev SI, Castle JC, Bukur V, Dietrich PY, Okada H, Walker PR. Responsiveness to anti-PD-1 and anti-CTLA-4 immune checkpoint blockade in SB28 and GL261 mouse glioma models. <i>Oncoimmunology</i> 2018;<i>7:</i>e1501137. 10.1080/2162402X.2018.1501137. Brennan CW, Verhaak RG, McKenna A, Campos B, Noushmehr H, Salama SR, Zheng S, Chakravarty D, Sanborn JZ, Berman SH, <i>et al.</i> The somatic genomic landscape of glioblastoma. <i>Cell</i> 2013;<i>155:</i>462–477. 10.1016/j.cell.2013.09.034.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.001 | 0.001 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.001 | 0.000 |
| Bibliometrics | 0.000 | 0.001 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.001 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".