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| Item Type: | Article |
|---|---|
| Title: | Personalized CRISPR knock-in cytokine gene therapy to remodel the tumor microenvironment and enhance CAR T cell therapy in solid tumors |
| Creators: |
Launspach, Michael  ORCID: https://orcid.org/0000-0001-5337-5605, Macos, Julia ORCID: https://orcid.org/0009-0008-9196-3975, Afzal, Shoaib ORCID: https://orcid.org/0009-0009-5490-6098, Hohmann, Janik ORCID: https://orcid.org/0009-0001-5374-1350, Appis, Marc L. ORCID: https://orcid.org/0000-0002-0155-1918, Pilgram, Maximilian ORCID: https://orcid.org/0009-0002-3423-5249, Beez, Stefanie, Ohlendorf, Emily, van der Ven, Casper F.T. ORCID: https://orcid.org/0000-0001-7947-6105, Lachiheb, Chahrazad, Töws, Karin, Andersch, Lena, Jens, Marvin ORCID: https://orcid.org/0000-0002-6660-8743, Zirngibl, Felix ORCID: https://orcid.org/0000-0003-1764-4841, Kath, Jonas ORCID: https://orcid.org/0000-0001-7721-7978, Stecklum, Maria ORCID: https://orcid.org/0009-0001-7951-5730, Rodriguez-Fos, Elias ORCID: https://orcid.org/0000-0002-2555-0178, Anders, Kathleen ORCID: https://orcid.org/0000-0002-0429-200X, Wagner, Dimitrios L. ORCID: https://orcid.org/0000-0002-2189-3579, Henssen, Anton G. ORCID: https://orcid.org/0000-0003-1534-778X, Kühn, Ralf ORCID: https://orcid.org/0000-0003-1694-9803, Eggert, Angelika and Künkele, Annette ORCID: https://orcid.org/0000-0002-8406-5412
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| Abstract: | The immunosuppressive tumour microenvironment (TME) remains a central barrier to effective immunotherapy in solid tumours. We present a gene-therapeutic strategy that enables localized remodelling of the TME via tumour-intrinsic cytokine expression. Central to this approach is CancerPAM, a multi-omics bioinformatics pipeline that identifies and ranks patient-specific, tumour-exclusive CRISPR-Cas9 knock-in sites with high specificity and integration efficiency. Using neuroblastoma as a model, CancerPAM analysis of tumour sequencing data identifies optimal knock-in sites for pro-inflammatory cytokines (CXCL10, CXCL11, IFNG), and CancerPAM rankings correlate strongly with target-site specificity and knock-in efficiency, validating its predictive performance. CRISPR-mediated CXCL10 knock-in enhances CAR T cell infiltration and antitumour efficacy in vitro and in vivo, including humanized CD34⁺ HuNOG mice, where CXCL10-expressing tumours show stronger immune infiltration and prolonged tumour control within a reconstituted human immune microenvironment. Our findings establish a framework for safe and effective CRISPR-based cytokine delivery, integrating localized TME remodelling with cellular immunotherapies to enhance CAR T cells and other treatments in immune-refractory solid tumours. |
| Keywords: | Adoptive Immunotherapy, CRISPR-Cas Systems, Chemokine CXCL10, Chemokine CXCL11, Chimeric Antigen Receptors, Clustered Regularly Interspaced Short Palindromic Repeats, Cytokines, Gene Knock-In Techniques, Genetic Therapy, Interferon-Gamma, Neoplasms, Neuroblastoma, Precision Medicine, T-Lymphocytes, Tumor Cell Line, Tumor Microenvironment, Animals, Mice |
| Source: | Nature Communications |
| ISSN: | 2041-1723 |
| Publisher: | Nature Publishing Group |
| Volume: | 16 |
| Number: | 1 |
| Page Range: | 10987 |
| Date: | 9 December 2025 |
| Official Publication: | https://doi.org/10.1038/s41467-025-67328-w |
| PubMed: | View item in PubMed |
| Related to: |
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