In Situ Modification of Tissue Stem and Progenitor Cell Genomes

Jill M Goldstein; Mohammadsharif Tabebordbar; Kexian Zhu; Leo D Wang; Kathleen A Messemer; Bryan Peacker; Sara Ashrafi Kakhki; Meryem Gonzalez-Celeiro; Yulia Shwartz; Jason K W Cheng; Ru Xiao; Trisha Barungi; Charles Albright; Ya-Chieh Hsu; Luk H Vandenberghe; Amy J Wagers

doi:10.1016/j.celrep.2019.03.105

In Situ Modification of Tissue Stem and Progenitor Cell Genomes

Cell Rep. 2019 Apr 23;27(4):1254-1264.e7. doi: 10.1016/j.celrep.2019.03.105.

Authors

Jill M Goldstein¹, Mohammadsharif Tabebordbar², Kexian Zhu³, Leo D Wang⁴, Kathleen A Messemer¹, Bryan Peacker¹, Sara Ashrafi Kakhki¹, Meryem Gonzalez-Celeiro⁵, Yulia Shwartz⁶, Jason K W Cheng⁷, Ru Xiao⁸, Trisha Barungi⁸, Charles Albright⁷, Ya-Chieh Hsu⁶, Luk H Vandenberghe⁹, Amy J Wagers¹⁰

Affiliations

¹ Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA; Paul F. Glenn Center for the Biology of Aging, Harvard Medical School, Boston, MA 02115, USA.
² Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
³ Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA; Paul F. Glenn Center for the Biology of Aging, Harvard Medical School, Boston, MA 02115, USA; Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA 02138, USA.
⁴ Joslin Diabetes Center, Boston, MA 02215, USA; Division of Pediatric Hematology/Oncology, Boston Children's Hospital and Dana-Farber Cancer Institute, Boston, MA 02215, USA.
⁵ Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA; Institute of Molecular Health Sciences, ETH Zurich, 8093 Zurich, Switzerland.
⁶ Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA.
⁷ Editas Medicine, Inc., 11 Hurley Street, Cambridge, MA 02142, USA.
⁸ Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, Boston, MA 02114, USA.
⁹ Harvard Stem Cell Institute, Cambridge, MA 02138, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear, Boston, MA 02114, USA; Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, Boston, MA 02114, USA.
¹⁰ Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA; Paul F. Glenn Center for the Biology of Aging, Harvard Medical School, Boston, MA 02115, USA; Joslin Diabetes Center, Boston, MA 02215, USA. Electronic address: amy_wagers@harvard.edu.

Abstract

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.

Keywords: AAV; dermal; gene modification; hematopoietic; mesenchymal; muscle; progenitor; satellite cell; stem cell.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Differentiation*
Cell Movement
Dependovirus / classification
Dependovirus / genetics*
Female
Gene Transfer Techniques
Genetic Therapy
Genome*
Hematopoietic Stem Cells / cytology*
Hematopoietic Stem Cells / metabolism
Humans
Male
Mice, Inbred C57BL
Mice, Inbred mdx
Satellite Cells, Skeletal Muscle / cytology*
Satellite Cells, Skeletal Muscle / metabolism
Skin / cytology*
Skin / metabolism

Abstract

Publication types

MeSH terms

Grants and funding