High-Efficiency, Selection-free Gene Repair in Airway Stem Cells from Cystic Fibrosis Patients Rescues CFTR Function in Differentiated Epithelia

Sriram Vaidyanathan; Ameen A. Salahudeen; Zachary M. Sellers; Dawn T. Bravo; Shannon S. Choi; Arpit Batish; Wei Le; Ron Baik; Sean de la O; Milan P. Kaushik; Noah Galper; Ciaran M. Lee; Christopher A. Teran; Jessica H. Yoo; Gang Bao; Eugene H. Chang; Zara M. Patel; Peter H. Hwang; Jeffrey J. Wine; Carlos E. Milla; Tushar J. Desai; Jayakar V. Nayak; Calvin J. Kuo; Matthew H. Porteus

doi:10.1016/j.stem.2019.11.002

High-Efficiency, Selection-free Gene Repair in Airway Stem Cells from Cystic Fibrosis Patients Rescues CFTR Function in Differentiated Epithelia

Sriram Vaidyanathan, Ameen A. Salahudeen, Zachary M. Sellers, Dawn T. Bravo, Shannon S. Choi, Arpit Batish, Wei Le, Ron Baik, Sean de la O, Milan P. Kaushik, Noah Galper, Ciaran M. Lee, Christopher A. Teran, Jessica H. Yoo, Gang Bao, Eugene H. Chang, Zara M. Patel, Peter H. Hwang, Jeffrey J. Wine, Carlos E. MillaTushar J. Desai, Jayakar V. Nayak, Calvin J. Kuo, Matthew H. Porteus

Otolaryngology

Research output: Contribution to journal › Article › peer-review

102 Scopus citations

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.

Original language	English (US)
Pages (from-to)	161-171.e4
Journal	Cell Stem Cell
Volume	26
Issue number	2
DOIs	https://doi.org/10.1016/j.stem.2019.11.002
State	Published - Feb 6 2020

Keywords

CF
CFTR
CRISPR
Cas9
F508del
airway stem cells
basal cells
cell therapy
cystic fibrosis
genome editing

ASJC Scopus subject areas

Molecular Medicine
Genetics
Cell Biology

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10.1016/j.stem.2019.11.002

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Vaidyanathan, S., Salahudeen, A. A., Sellers, Z. M., Bravo, D. T., Choi, S. S., Batish, A., Le, W., Baik, R., de la O, S., Kaushik, M. P., Galper, N., Lee, C. M., Teran, C. A., Yoo, J. H., Bao, G., Chang, E. H., Patel, Z. M., Hwang, P. H., Wine, J. J., ... Porteus, M. H. (2020). High-Efficiency, Selection-free Gene Repair in Airway Stem Cells from Cystic Fibrosis Patients Rescues CFTR Function in Differentiated Epithelia. Cell Stem Cell, 26(2), 161-171.e4. https://doi.org/10.1016/j.stem.2019.11.002

Vaidyanathan, S, Salahudeen, AA, Sellers, ZM, Bravo, DT, Choi, SS, Batish, A, Le, W, Baik, R, de la O, S, Kaushik, MP, Galper, N, Lee, CM, Teran, CA, Yoo, JH, Bao, G, Chang, EH, Patel, ZM, Hwang, PH, Wine, JJ, Milla, CE, Desai, TJ, Nayak, JV, Kuo, CJ & Porteus, MH 2020, 'High-Efficiency, Selection-free Gene Repair in Airway Stem Cells from Cystic Fibrosis Patients Rescues CFTR Function in Differentiated Epithelia', Cell Stem Cell, vol. 26, no. 2, pp. 161-171.e4. https://doi.org/10.1016/j.stem.2019.11.002

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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.}",

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AU - Vaidyanathan, Sriram

AU - Salahudeen, Ameen A.

AU - Sellers, Zachary M.

AU - Bravo, Dawn T.

AU - Choi, Shannon S.

AU - Batish, Arpit

AU - Le, Wei

AU - Baik, Ron

AU - de la O, Sean

AU - Kaushik, Milan P.

AU - Galper, Noah

AU - Lee, Ciaran M.

AU - Teran, Christopher A.

AU - Yoo, Jessica H.

AU - Bao, Gang

AU - Chang, Eugene H.

AU - Patel, Zara M.

AU - Hwang, Peter H.

AU - Wine, Jeffrey J.

AU - Milla, Carlos E.

AU - Desai, Tushar J.

AU - Nayak, Jayakar V.

AU - Kuo, Calvin J.

AU - Porteus, Matthew H.

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N2 - 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.

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High-Efficiency, Selection-free Gene Repair in Airway Stem Cells from Cystic Fibrosis Patients Rescues CFTR Function in Differentiated Epithelia

Abstract

Keywords

ASJC Scopus subject areas

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