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| Item Type: | Preprint |
|---|---|
| Title: | Personalized CRISPR knock-in cytokine gene therapy to remodel the tumor microenvironment and enhance CAR T cell therapy in solid tumors |
| Creators Name: | Launspach, M., Macos, J., Afzal, S., Hohmann, J., Beez, S., Pilgram, M., Ohlendorf, E., Yevchuk, A., van der Ven, C.F.T., Lachiheb, C., Andersch, L., Jens, M., Appis, M.L., Zirngibl, F., Kath, J., Stecklum, M., Anders, K., Wagner, D.L., Kühn, R., Eggert, A. and Künkele, A. |
| Abstract: | The immunosuppressive tumor microenvironment (TME) remains a central barrier to effective immunotherapy in solid tumors. To address this, we developed a novel gene therapeutic strategy that enables localized remodeling of the TME via tumor-intrinsic cytokine expression. Central to this approach is CancerPAM, a multi-omics bioinformatics pipeline that identifies and ranks patient-specific, tumor-exclusive CRISPR-Cas9 knock-in sites with high specificity and integration efficiency. Using neuroblastoma—a pediatric solid tumor with a suppressive TME—as a model, we applied CancerPAM to sequencing data from cell lines and patients to identify optimal integration sites for pro-inflammatory cytokines (CXCL10, CXCL11, IFNG). CRISPR-mediated CXCL10 knock-in into tumor cells significantly enhanced CAR T cell infiltration and antitumor efficacy both in vitro and in vivo. In vivo, CXCL10expressing tumors showed significantly increased early CAR T cell infiltration and prolonged survival compared to controls. CancerPAM rankings correlated strongly with target-site specificity and knock-in efficiency, validating its predictive performance. Our findings establish CancerPAM as a powerful tool for safe and effective CRISPR-based interventions and provide a conceptual framework for integrating cytokine-driven TME remodeling with cellular immunotherapies. This personalized strategy holds promise for enhancing CAR T cells and other immunotherapies across immune-refractory solid tumors. |
| Keywords: | Animals, Mice |
| Source: | bioRxiv |
| Publisher: | Cold Spring Harbor Laboratory Press |
| Article Number: | 2025.04.25.650703 |
| Date: | 27 April 2025 |
| Official Publication: | https://doi.org/10.1101/2025.04.25.650703 |
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