Journal article
Targeted replacement of full-length CFTR in human airway stem cells by CRISPR-Cas9 for pan-mutation correction in the endogenous locus
Molecular therapy, Vol.30(1), pp.223-237
01/05/2022
DOI: 10.1016/j.ymthe.2021.03.023
PMCID: PMC8753290
PMID: 33794364
Abstract
Cystic fibrosis (CF) is a monogenic disease caused by impaired production and/or function of the CF transmembrane conductance regulator (CFTR) protein. Although we have previously shown correction of the most common pathogenic mutation, there are many other pathogenic mutations throughout the CF gene. An autologous airway stem cell therapy in which the CFTR cDNA is precisely inserted into the CFTR locus may enable the development of a durable cure for almost all CF patients, irrespective of the causal mutation. Here, we use CRISPR-Cas9 and two adeno-associated viruses (AAVs) carrying the two halves of the CFTR cDNA to sequentially insert the full CFTR cDNA along with a truncated CD19 (tCD19) enrichment tag in upper airway basal stem cells (UABCs) and human bronchial epithelial cells (HBECs). The modified cells were enriched to obtain 60%-80% tCD19+ UABCs and HBECs from 11 different CF donors with a variety of mutations. Differentiated epithelial monolayers cultured at air-liquid the CFTR function in non-CF controls. Thus, our study enables the development of a therapy for almost all CF patients, including patients who cannot be treated using recently approved modulator therapies.
Details
- Title: Subtitle
- Targeted replacement of full-length CFTR in human airway stem cells by CRISPR-Cas9 for pan-mutation correction in the endogenous locus
- Creators
- Sriram Vaidyanathan - Stanford UniversityRon Baik - Stanford UniversityLu Chen - Stanford UniversityDawn T. Bravo - Dept Otolaryngol Head & Neck Surg, Stanford, CA 94305 USACarlos J. Suarez - Stanford UniversityShayda M. Abazari - Stanford UniversityAmeen A. Salahudeen - Stanford UniversityAmanda M. Dudek - Stanford UniversityChristopher A. Teran - Dept Otolaryngol Head & Neck Surg, Stanford, CA 94305 USATimothy H. Davis - Rice UniversityCiaran M. Lee - Rice UniversityGang Bao - Rice UniversityScott H. Randell - Univ N Carolina, Dept Cell Biol & Physiol, Chapel Hill, NC 27599 USASteven E. Artandi - Stanford UniversityJeffrey J. Wine - Stanford UniversityCalvin J. Kuo - Stanford UniversityTushar J. Desai - Stanford UniversityJayakar Nayak - Dept Otolaryngol Head & Neck Surg, Stanford, CA 94305 USAZachary M. Sellers - Stanford UniversityMatthew H. Porteus - Stanford University
- Resource Type
- Journal article
- Publication Details
- Molecular therapy, Vol.30(1), pp.223-237
- DOI
- 10.1016/j.ymthe.2021.03.023
- PMID
- 33794364
- PMCID
- PMC8753290
- NLM abbreviation
- Mol Ther
- ISSN
- 1525-0016
- eISSN
- 1525-0024
- Publisher
- Elsevier
- Number of pages
- 15
- Grant note
- R35CA197563; U01CA217851 / National Cancer Institute; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Cancer Institute (NCI) R01HL151677 / National Heart Lung and Blood Institute; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Heart Lung & Blood Institute (NHLBI) R01AG056575 / National Institute on Aging; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute on Aging (NIA) BOUCHE19RO / CF Foundation RR14008 / Cancer Prevention and Research Institute of Texas; Cancer Prevention & Research Institute of Texas Stanford-SPARK MCHRI K08DK124684; K08DE027730; U01DK085527; U01CA217851; U01CA176299; U19AI116484; DK065988 / NIH; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA Crandall Foundation; ACEV Foundation DISC2-09637 / California Institute of Regenerative Medicine; California Institute for Regenerative Medicine BAO19XX0; VAIDYA19F0; SELLER16L0 / Cystic Fibrosis Foundation; Italian Cystic Fibrosis Research Foundation
- Language
- English
- Date published
- 01/05/2022
- Academic Unit
- Microbiology and Immunology
- Record Identifier
- 9984822990802771
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