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Cas9-AAV6-engineered human mesenchymal stromal cells improved cutaneous wound healing in diabetic mice

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 mutag...

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Autores principales: Srifa, Waracharee, Kosaric, Nina, Amorin, Alvaro, Jadi, Othmane, Park, Yujin, Mantri, Sruthi, Camarena, Joab, Gurtner, Geoffrey C., Porteus, Matthew
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7235221/
https://www.ncbi.nlm.nih.gov/pubmed/32424320
http://dx.doi.org/10.1038/s41467-020-16065-3
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author Srifa, Waracharee
Kosaric, Nina
Amorin, Alvaro
Jadi, Othmane
Park, Yujin
Mantri, Sruthi
Camarena, Joab
Gurtner, Geoffrey C.
Porteus, Matthew
author_facet Srifa, Waracharee
Kosaric, Nina
Amorin, Alvaro
Jadi, Othmane
Park, Yujin
Mantri, Sruthi
Camarena, Joab
Gurtner, Geoffrey C.
Porteus, Matthew
author_sort Srifa, Waracharee
collection PubMed
description 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.
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spelling pubmed-72352212020-05-20 Cas9-AAV6-engineered human mesenchymal stromal cells improved cutaneous wound healing in diabetic mice Srifa, Waracharee Kosaric, Nina Amorin, Alvaro Jadi, Othmane Park, Yujin Mantri, Sruthi Camarena, Joab Gurtner, Geoffrey C. Porteus, Matthew Nat Commun Article 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. Nature Publishing Group UK 2020-05-18 /pmc/articles/PMC7235221/ /pubmed/32424320 http://dx.doi.org/10.1038/s41467-020-16065-3 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Srifa, Waracharee
Kosaric, Nina
Amorin, Alvaro
Jadi, Othmane
Park, Yujin
Mantri, Sruthi
Camarena, Joab
Gurtner, Geoffrey C.
Porteus, Matthew
Cas9-AAV6-engineered human mesenchymal stromal cells improved cutaneous wound healing in diabetic mice
title Cas9-AAV6-engineered human mesenchymal stromal cells improved cutaneous wound healing in diabetic mice
title_full Cas9-AAV6-engineered human mesenchymal stromal cells improved cutaneous wound healing in diabetic mice
title_fullStr Cas9-AAV6-engineered human mesenchymal stromal cells improved cutaneous wound healing in diabetic mice
title_full_unstemmed Cas9-AAV6-engineered human mesenchymal stromal cells improved cutaneous wound healing in diabetic mice
title_short Cas9-AAV6-engineered human mesenchymal stromal cells improved cutaneous wound healing in diabetic mice
title_sort cas9-aav6-engineered human mesenchymal stromal cells improved cutaneous wound healing in diabetic mice
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7235221/
https://www.ncbi.nlm.nih.gov/pubmed/32424320
http://dx.doi.org/10.1038/s41467-020-16065-3
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