In Situ Modification of Tissue Stem and Progenitor Cell Genomes

• Published in: Cell reports (Cambridge) Vol. 27; no. 4; pp. 1254 - 1264.e7
• Main Authors: Goldstein, Jill M., Tabebordbar, Mohammadsharif, Zhu, Kexian, Wang, Leo D., Messemer, Kathleen A., Peacker, Bryan, Ashrafi Kakhki, Sara, Gonzalez-Celeiro, Meryem, Shwartz, Yulia, Cheng, Jason K.W., Xiao, Ru, Barungi, Trisha, Albright, Charles, Hsu, Ya-Chieh, Vandenberghe, Luk H., Wagers, Amy J.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 23.04.2019
• Subjects: AAV; Animals; Cell Differentiation; Cell Movement; Dependovirus - classification; Dependovirus - genetics; dermal; Female; gene modification; Gene Transfer Techniques; Genetic Therapy; Genome; hematopoietic; Hematopoietic Stem Cells - cytology; Hematopoietic Stem Cells - metabolism; Humans; Male; mesenchymal; Mice, Inbred C57BL; Mice, Inbred mdx; muscle; progenitor; satellite cell; Satellite Cells, Skeletal Muscle - cytology; Satellite Cells, Skeletal Muscle - metabolism; Skin - cytology; Skin - metabolism; stem cell; satellite cell; hematopoietic; stem cell; mesenchymal; AAV; muscle; gene modification; progenitor; dermal
• ISSN: 2211-1247; 2211-1247
• DOI: 10.1016/j.celrep.2019.03.105

In vivo delivery of genome-modifying enzymes holds significant promise for therapeutic applications and functional genetic screening. Delivery to endogenous tissue stem cells, which provide an enduring source of cell replacement during homeostasis and regeneration, is of particular interest. Here, we use a sensitive Cre/lox fluorescent reporter system to test the efficiency of genome modification following in vivo transduction by adeno-associated viruses (AAVs) in tissue stem and progenitor cells. We combine immunophenotypic analyses with in vitro and in vivo assays of stem cell function to reveal effective targeting of skeletal muscle satellite cells, mesenchymal progenitors, hematopoietic stem cells, and dermal cell subsets using multiple AAV serotypes. Genome modification rates achieved through this system reached >60%, and modified cells retained key functional properties. This study establishes a powerful platform to genetically alter tissue progenitors within their physiological niche while preserving their native stem cell properties and regulatory interactions. [Display omitted] •Multiple AAV serotypes transduce tissue stem cells via systemic or local delivery•AAV-delivered Cre recombinase modifies stem cells in multiple anatomical niches•In vivo AAV transduction does not require stem cell isolation or transplantation•AAV-transduced stem cells retain differentiation and engraftment capacities Goldstein et al. demonstrate in vivo transduction of endogenous tissue stem cells in the muscle, blood, and skin by systemic or local administration of adeno-associated viruses (AAVs) encoding genome-modifying enzymes. They report that AAV-transduced and genome-modified stem and progenitor cells maintain their capacity to differentiate and engraft following transplantation.