@article{345341ba857840ab91279baebd293e81,
title = "High-Efficiency, Selection-free Gene Repair in Airway Stem Cells from Cystic Fibrosis Patients Rescues CFTR Function in Differentiated Epithelia",
abstract = "Cystic fibrosis (CF) is a monogenic disorder caused by mutations in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene. Mortality in CF patients is mostly due to respiratory sequelae. Challenges with gene delivery have limited attempts to treat CF using in vivo gene therapy, and low correction levels have hindered ex vivo gene therapy efforts. We have used Cas9 and adeno-associated virus 6 to correct the ΔF508 mutation in readily accessible upper-airway basal stem cells (UABCs) obtained from CF patients. On average, we achieved 30%–50% allelic correction in UABCs and bronchial epithelial cells (HBECs) from 10 CF patients and observed 20%–50% CFTR function relative to non-CF controls in differentiated epithelia. Furthermore, we successfully embedded the corrected UABCs on an FDA-approved porcine small intestinal submucosal membrane (pSIS), and they retained differentiation capacity. This study supports further development of genetically corrected autologous airway stem cell transplant as a treatment for CF.",
keywords = "CF, CFTR, CRISPR, Cas9, F508del, airway stem cells, basal cells, cell therapy, cystic fibrosis, genome editing",
author = "Sriram Vaidyanathan and Salahudeen, {Ameen A.} and Sellers, {Zachary M.} and Bravo, {Dawn T.} and Choi, {Shannon S.} and Arpit Batish and Wei Le and Ron Baik and {de la O}, Sean and Kaushik, {Milan P.} and Noah Galper and Lee, {Ciaran M.} and Teran, {Christopher A.} and Yoo, {Jessica H.} and Gang Bao and Chang, {Eugene H.} and Patel, {Zara M.} and Hwang, {Peter H.} and Wine, {Jeffrey J.} and Milla, {Carlos E.} and Desai, {Tushar J.} and Nayak, {Jayakar V.} and Kuo, {Calvin J.} and Porteus, {Matthew H.}",
note = "Funding Information: We thank Dr. Scott Randell (University of North Carolina) for guidance on Ussing chamber assays and training on ALI cultures. We thank Dr. Jeffrey Beekman and Dr. Gimano Amatngalim for their guidance on Ussing chamber assays. We appreciate the efforts of Ivan T. Lee, Nicole Borchard, Sachi Dholakia, David Zarabanda, Phillip Gall, and Yasuyuki Noyama in prioritizing efforts at human tissue acquisition from the operating theater. We thank the CF Canada Primary Airway Cell Biobank at McGill University for providing primary bronchial airway epithelial cells. CFTR antibodies were obtained from Dr. John Riordan at the University of North Carolina, Chapel Hill through the CF foundation antibody distribution program. The work was funded by grants from the California Institute of Regenerative Medicine, United States ( DISC2-09637 ), Stanford-SPARK MCHRI , and the Cancer Prevention and Research Institute of Texas, United States ( RR14008 and RP170721 ). We thank the Crandall Foundation for a philanthropic gift that supported this work. S.V. and Z.M.S. were supported by the Cystic Fibrosis Foundation, United States (awards: VAIDYA19F0, SELLER16L0). A.A.S. was supported by a fellowship from the A.P. Giannini Foundation , United States; the Eastern Cooperative Group Paul Carbone Award , and an NIDCR Career Development Award 1K08DE027730 . Funding Information: We thank Dr. Scott Randell (University of North Carolina) for guidance on Ussing chamber assays and training on ALI cultures. We thank Dr. Jeffrey Beekman and Dr. Gimano Amatngalim for their guidance on Ussing chamber assays. We appreciate the efforts of Ivan T. Lee, Nicole Borchard, Sachi Dholakia, David Zarabanda, Phillip Gall, and Yasuyuki Noyama in prioritizing efforts at human tissue acquisition from the operating theater. We thank the CF Canada Primary Airway Cell Biobank at McGill University for providing primary bronchial airway epithelial cells. CFTR antibodies were obtained from Dr. John Riordan at the University of North Carolina, Chapel Hill through the CF foundation antibody distribution program. The work was funded by grants from the California Institute of Regenerative Medicine, United States (DISC2-09637), Stanford-SPARK MCHRI, and the Cancer Prevention and Research Institute of Texas, United States (RR14008 and RP170721). We thank the Crandall Foundation for a philanthropic gift that supported this work. S.V. and Z.M.S. were supported by the Cystic Fibrosis Foundation, United States (awards: VAIDYA19F0, SELLER16L0). A.A.S. was supported by a fellowship from the A.P. Giannini Foundation, United States; the Eastern Cooperative Group Paul Carbone Award, and an NIDCR Career Development Award 1K08DE027730. Conceptualization, S.V. A.A.S. D.T.B. J.V.N. T.J.D. M.H.P. and C.J.K.; Investigation, S.V. A.A.S. Z.M.S. and D.T.B.; Writing, S.V. A.A.S. Z.M.S. D.T.B. M.H.P. J.V.N. T.J.D. and C.J.K.; S.S.C. A.B. W.L. R.B. S.d.l.O. M.P.K. N.G. C.M.L. C.A.T. J.H.Y. G.B. E.H.C. Z.M.P. P.H.H. J.J.W. and C.E.M. all participated in the design and conception of experiments and provided editorial feedback on the manuscript. M.H.P. has equity and serves on the scientific advisory board of CRISPR Therapeutics. J.V.N. is a consultant with COOK Medical, which manufactures the pSIS graft. Neither company had any input on the design, execution, interpretation, or publication of the work in this manuscript. Publisher Copyright: {\textcopyright} 2019 Elsevier Inc.",
year = "2020",
month = feb,
day = "6",
doi = "10.1016/j.stem.2019.11.002",
language = "English (US)",
volume = "26",
pages = "161--171.e4",
journal = "Cell Stem Cell",
issn = "1934-5909",
publisher = "Cell Press",
number = "2",
}