| Popis: |
The hostile tumor microenvironment (TME) is major challenge for cancer immunotherapies. Here, we design and perform TME-targetedin vivoCRISPR 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 encodingantigenpresentation,cytokine, and co-stimulationmolecules (APCM) leads to enhanced anti-tumor immunity. APCM screen in metastatic tumors identifiesCd80, Tnfsf14, Cxcl10, Tnfsf18, Tnfsf9, andIfngas the top immunostimulatory candidates. AAV-mediated delivery of these factors individually or in combination shows anti-tumor efficacy across different cancer models. Further optimization pinpointsIfng+Tnfsf9+Il12b(Il12/Il23)as a potent therapeutic combination, leading to increased IFN-γ+CD8+and tissue-resident memory T cells. APCM therapy synergizes with CAR-T cell therapy against human solid tumorsin 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. |
