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bioRxiv · 10.1101/2023.03.14.532665

Rational design of immune gene therapy combinations via in vivo CRISPR activation screen of tumor microenvironment modulators

Wang, G.·Zhang, F.·Chow, R. D.·He, E.·Zhu, L.·Han, Q.·Chen, S.
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Abstract

The hostile tumor microenvironment (TME) is major challenge for cancer immunotherapies. Here, we design and perform TME-targeted in vivo CRISPR activation (CRISPRa) screens to uncover factors that promote anti-tumor immunity, culminating in rationally designed immune gene therapy combinations. Through adeno-associated virus (AAV) delivery, multiplexed activation of pooled immunoregulatory genes encoding antigen presentation, cytokine, and co-stimulation molecules (APCM) leads to enhanced anti-tumor immunity. APCM screen in metastatic tumors identifies Cd80, Tnfsf14, Cxcl10, Tnfsf18, Tnfsf9, and Ifng as the top immunostimulatory candidates. AAV-mediated delivery of these factors individually or in combination shows anti-tumor efficacy across different cancer models. Further optimization pinpoints Ifng+Tnfsf9+Il12b(Il12/Il23) as a potent therapeutic combination, leading to increased IFN-{gamma}+CD8+ and tissue-resident memory T cells. APCM therapy synergizes with CAR-T cell therapy against human solid tumors in vivo. APCM-based CRISPRa screen and gene activation systems can thus be leveraged for the rapid generation of off-the-shelf immune gene therapies against solid tumors.

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Wang, G., Zhang, F., Chow, R. D., He, E., Zhu, L., Han, Q., Chen, S.. 2023-03-15. Rational design of immune gene therapy combinations via in vivo CRISPR activation screen of tumor microenvironment modulators. https://doi.org/10.1101/2023.03.14.532665

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