Base Editor Correction of COL7A1 in Recessive Dystrophic Epidermolysis Bullosa Patient-Derived Fibroblasts and iPSCs

Mark J. Osborn, Gregory A. Newby, Amber N. McElroy, Friederike Knipping, Sarah C. Nielsen, Megan J. Riddle, Lily Xia, Weili Chen, Cindy R. Eide, Beau R. Webber, Hans H. Wandall, Sally Dabelsteen, Bruce R. Blazar, David R. Liu, Jakub Tolar

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

Abstract

Genome editing represents a promising strategy for the therapeutic correction of COL7A1 mutations that cause recessive dystrophic epidermolysis bullosa (RDEB). DNA cleavage followed by homology-directed repair (HDR) using an exogenous template has previously been used to correct COL7A1 mutations. HDR rates can be modest, and the double-strand DNA breaks that initiate HDR commonly result in accompanying undesired insertions and deletions (indels). To overcome these limitations, we applied an A•T→G•C adenine base editor (ABE) to correct two different COL7A1 mutations in primary fibroblasts derived from RDEB patients. ABE enabled higher COL7A1 correction efficiencies than previously reported HDR efforts. Moreover, ABE obviated the need for a repair template, and minimal indels or editing at off-target sites was detected. Base editing restored the endogenous type VII collagen expression and function in vitro. We also treated induced pluripotent stem cells (iPSCs) derived from RDEB fibroblasts with ABE. The edited iPSCs were differentiated into mesenchymal stromal cells, a cell population with therapeutic potential for RDEB. In a mouse teratoma model, the skin derived from ABE-treated iPSCs showed the proper deposition of C7 at the dermal–epidermal junction in vivo. These demonstrate that base editing provides an efficient and precise genome editing method for autologous cell engineering for RDEB.

Original languageEnglish (US)
Pages (from-to)338-347.e5
JournalJournal of Investigative Dermatology
Volume140
Issue number2
DOIs
StatePublished - Feb 2020

Bibliographical note

Funding Information:
This work was supported by funding from the Mighty Mimi St. Baldrick's Foundation Scholar Award, the Richard M. Schulze Family Foundation, US NIH R01 AR063070, R01 HL56067, DebRA International, Jackson Gabriel Silver Fund, Epidermolysis Bullosa Medical Research Fund, Kidz1stFund, U.S. NIH U01 AI142756, RM1 HG009490, and R35 GM118062, the St. Jude's Collaborative Research Consortium, the Helen Hay Whitney Foundation, and the Howard Hughes Medical Institute. The authors are grateful to Drs Mei Chen & David Woodley for the C7 antibody and Dr James Rheinwald for organotypic culture keratinocytes. Conceptualization: MJO, GAN, BRB, DRL, JT; Methodology: MJO, GAN, FK, ANM, SCN, MJR, LX, WC, CRE, BRW, SD.; Formal Analysis: MJO, GAN, FK; Funding Acquisition: MJO, GAN, HHW, SD, BRB, DRL, JT; Investigation: MJO, GAN, FK, ANM, SCN, MJR, LX, WC, SD; Resources: MJO, GAN, HHW, BRB, SD, DRL, JT.; Writing: MJO, GAN, ANM, FK, SCN, MJR, LX, WC, CRE, BRW, HHW, SD, BRB, DRL, and JT.

Funding Information:
This work was supported by funding from the Mighty Mimi St. Baldrick’s Foundation Scholar Award, the Richard M. Schulze Family Foundation , US NIH R01 AR063070 , R01 HL56067 , DebRA International, Jackson Gabriel Silver Fund, Epidermolysis Bullosa Medical Research Fund, Kidz1stFund , U.S. NIH U01 AI142756 , RM1 HG009490 , and R35 GM118062 , the St. Jude’s Collaborative Research Consortium , the Helen Hay Whitney Foundation , and the Howard Hughes Medical Institute . The authors are grateful to Drs Mei Chen & David Woodley for the C7 antibody and Dr James Rheinwald for organotypic culture keratinocytes.

Publisher Copyright:
© 2019 The Authors

PubMed: MeSH publication types

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

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