Cas9-AAV6-engineered human mesenchymal stromal cells improved cutaneous wound healing in diabetic mice.
Animals
CRISPR-Associated Protein 9
/ metabolism
Cell Proliferation
Cell Survival
Cross-Linking Reagents
/ chemistry
Dependovirus
Diabetes Mellitus, Experimental
/ pathology
Gene Editing
Green Fluorescent Proteins
/ metabolism
Humans
Hydrogels
/ chemistry
Kinetics
Mesenchymal Stem Cell Transplantation
Mesenchymal Stem Cells
/ metabolism
Mice
Proto-Oncogene Proteins c-sis
Skin
/ pathology
Vascular Endothelial Growth Factor A
/ metabolism
Wound Healing
Xenograft Model Antitumor Assays
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
18 05 2020
18 05 2020
Historique:
received:
29
07
2019
accepted:
25
02
2020
entrez:
20
5
2020
pubmed:
20
5
2020
medline:
13
8
2020
Statut:
epublish
Résumé
Human mesenchymal stromal cells (hMSCs) are a promising source for engineered cell-based therapies in which genetic engineering could enhance therapeutic efficacy and install novel cellular functions. Here, we describe an optimized Cas9-AAV6-based genome editing tool platform for site-specific mutagenesis and integration of up to more than 3 kilobases of exogenous DNA in the genome of hMSCs derived from the bone marrow, adipose tissue, and umbilical cord blood without altering their ex vivo characteristics. We generate safe harbor-integrated lines of engineered hMSCs and show that engineered luciferase-expressing hMSCs are transiently active in vivo in wound beds of db/db mice. Moreover, we generate PDGF-BB- and VEGFA-hypersecreting hMSC lines as short-term, local wound healing agents with superior therapeutic efficacy over wildtype hMSCs in the diabetic mouse model without replacing resident cells long-term. This study establishes a precise genetic engineering platform for genetic studies of hMSCs and development of engineered hMSC-based therapies.
Identifiants
pubmed: 32424320
doi: 10.1038/s41467-020-16065-3
pii: 10.1038/s41467-020-16065-3
pmc: PMC7235221
doi:
Substances chimiques
Cross-Linking Reagents
0
Hydrogels
0
Proto-Oncogene Proteins c-sis
0
Vascular Endothelial Growth Factor A
0
Green Fluorescent Proteins
147336-22-9
CRISPR-Associated Protein 9
EC 3.1.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2470Subventions
Organisme : NIDDK NIH HHS
ID : R01 DK074095
Pays : United States
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