Cas9-mediated knockout of Ndrg2 enhances the regenerative potential of dendritic cells for wound healing

• Published in: Nature communications 2023-08, Vol.14 (1), p.4729-16, Article 4729
• Main Authors: Henn, Dominic, Zhao, Dehua, Sivaraj, Dharshan, Trotsyuk, Artem, Bonham, Clark Andrew, Fischer, Katharina S., Kehl, Tim, Fehlmann, Tobias, Greco, Autumn H., Kussie, Hudson C., Moortgat Illouz, Sylvia E., Padmanabhan, Jagannath, Barrera, Janos A., Kneser, Ulrich, Lenhof, Hans-Peter, Januszyk, Michael, Levi, Benjamin, Keller, Andreas, Longaker, Michael T., Chen, Kellen, Qi, Lei S., Gurtner, Geoffrey C.
• Format: Article
• Language: English
• Subjects: 14/19; 38/1; 38/23; 38/39; 38/91; 42/41; 45; 45/62; 631/208/4041/3196; 631/61/490; 692/308/575; Angiogenesis; Animal models; Animals; Clinical trials; Craniocerebral Trauma; CRISPR; CRISPR-Cas Systems; Dendritic Cells; Dendritic structure; Diabetes; Diabetes mellitus; Gene Editing; Gene sequencing; Genes, myc; Genetic modification; Genome editing; Humanities and Social Sciences; Humans; Hydrogels; Immunotherapy; Mice; multidisciplinary; Myc protein; Patients; Progenitor cells; Science; Science (multidisciplinary); Transcriptomics; Wound healing; Wound Healing - genetics
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-023-40519-z

Chronic wounds impose a significant healthcare burden to a broad patient population. Cell-based therapies, while having shown benefits for the treatment of chronic wounds, have not yet achieved widespread adoption into clinical practice. We developed a CRISPR/Cas9 approach to precisely edit murine dendritic cells to enhance their therapeutic potential for healing chronic wounds. Using single-cell RNA sequencing of tolerogenic dendritic cells, we identified N-myc downregulated gene 2 ( Ndrg2 ), which marks a specific population of dendritic cell progenitors, as a promising target for CRISPR knockout. Ndrg2 -knockout alters the transcriptomic profile of dendritic cells and preserves an immature cell state with a strong pro-angiogenic and regenerative capacity. We then incorporated our CRISPR-based cell engineering within a therapeutic hydrogel for in vivo cell delivery and developed an effective translational approach for dendritic cell-based immunotherapy that accelerated healing of full-thickness wounds in both non-diabetic and diabetic mouse models. These findings could open the door to future clinical trials using safe gene editing in dendritic cells for treating various types of chronic wounds. Chronic wounds impose a significant burden to a broad patient population. Here, the authors use CRISPR/Cas9 to enhance the regenerative capacity of dendritic cells by knocking out the gene Ndrg2, and show that seeding these engineered dendritic cells on hydrogels constitutes an effective therapy for chronic wounds in diabetic and non-diabetic conditions.