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

Research output: Contribution to journalArticlepeer-review

43 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 languageEnglish (US)
Pages (from-to)161-171.e4
JournalCell Stem Cell
Volume26
Issue number2
DOIs
StatePublished - Feb 6 2020
Externally publishedYes

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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