Targeted correction and restored function of the CFTR gene in cystic fibrosis induced pluripotent stem cells

Ana M. Crane; Philipp Kramer; Jacquelin H. Bui; Wook Joon Chung; Xuan Shirley Li; Manuel L. Gonzalez-Garay; Finn Hawkins; Wei Liao; Daniela Mora; Sangbum Choi; Jianbin Wang; Helena C. Sun; David E. Paschon; Dmitry Y. Guschin; Philip D. Gregory; Darrell N. Kotton; Michael C. Holmes; Eric J. Sorscher; Brian R. Davis

doi:10.1016/j.stemcr.2015.02.005

Targeted correction and restored function of the CFTR gene in cystic fibrosis induced pluripotent stem cells

Ana M. Crane
, Philipp Kramer
, Jacquelin H. Bui
, Wook Joon Chung
, Xuan Shirley Li
, Manuel L. Gonzalez-Garay
, Finn Hawkins
, Wei Liao
, Daniela Mora
, Sangbum Choi
, Jianbin Wang
, Helena C. Sun
, David E. Paschon
, Dmitry Y. Guschin
, Philip D. Gregory
, Darrell N. Kotton
, Michael C. Holmes
, Eric J. Sorscher
, Brian R. Davis^*

^*Corresponding author for this work

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

175 Citations (Scopus)

Abstract

Recently developed reprogramming and genome editing technologies make poßible the derivation of corrected patient-specific pluripotent stem cell sources-potentially useful for the development of new therapeutic approaches. Starting with skin fibroblasts from patients diagnosed with cystic fibrosis, we derived and characterized induced pluripotent stem cell (iPSC) lines.We then utilized zinc-finger nucleases (ZFNs), designed to target the endogenous CFTR gene, to mediate correction of the inherited genetic mutation in these patientderived lines via homology-directed repair (HDR). We observed an exquisitely sensitive, homology-dependent preference for targeting one CFTR allele versus the other. The corrected cystic fibrosis iPSCs, when induced to differentiate in vitro, expreßed the corrected CFTR gene; importantly, CFTR correction resulted in restored expreßion of the mature CFTR glycoprotein and restoration of CFTR chloride channel function in iPSC-derived epithelial cells.

Original language	English
Pages (from-to)	569-577
Number of pages	9
Journal	Stem Cell Reports
Volume	4
Issue number	4
DOIs	https://doi.org/10.1016/j.stemcr.2015.02.005
Publication status	Published - 2015
Externally published	Yes

Bibliographical note

Funding Information:
We thank Naoki Nakayama (Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center) for helpful discussions; Veronica K. Quiceno for editorial assistance; Kathryn Plath, Connie Cepko, Brian Sauer, and Jeffrey Beekman for providing DNA constructs either directly or through Addgene; and Fyodor Urnov (Sangamo BioSciences, Inc.) for ZFN development through funding from the Cystic Fibrosis Foundation (CFF) Folding Consortium. This study was supported by NIH RC1HL099559 (B.R.D. and R. Wetsel, primary investigators [PIs]); CFF DAVIS12GO (B.R.D., PI); NIH P30 DK072482 (E.J.S., PI); and CFF R464 (E.J.S., PI). J.W., H.C.S., D.E.P., D.Y.G, P.D.G., and M.C.H. are employees of Sangamo BioSciences, Inc.

ASJC Scopus subject areas

Biochemistry
Genetics
Developmental Biology
Cell Biology

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10.1016/j.stemcr.2015.02.005

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Crane, A. M., Kramer, P., Bui, J. H., Chung, W. J., Li, X. S., Gonzalez-Garay, M. L., Hawkins, F., Liao, W., Mora, D., Choi, S., Wang, J., Sun, H. C., Paschon, D. E., Guschin, D. Y., Gregory, P. D., Kotton, D. N., Holmes, M. C., Sorscher, E. J., & Davis, B. R. (2015). Targeted correction and restored function of the CFTR gene in cystic fibrosis induced pluripotent stem cells. Stem Cell Reports, 4(4), 569-577. https://doi.org/10.1016/j.stemcr.2015.02.005

Crane, AM, Kramer, P, Bui, JH, Chung, WJ, Li, XS, Gonzalez-Garay, ML, Hawkins, F, Liao, W, Mora, D, Choi, S, Wang, J, Sun, HC, Paschon, DE, Guschin, DY, Gregory, PD, Kotton, DN, Holmes, MC, Sorscher, EJ & Davis, BR 2015, 'Targeted correction and restored function of the CFTR gene in cystic fibrosis induced pluripotent stem cells', Stem Cell Reports, vol. 4, no. 4, pp. 569-577. https://doi.org/10.1016/j.stemcr.2015.02.005

@article{0328e170ba13441fb443c6753f5e429a,

title = "Targeted correction and restored function of the CFTR gene in cystic fibrosis induced pluripotent stem cells",

abstract = "Recently developed reprogramming and genome editing technologies make po{\ss}ible the derivation of corrected patient-specific pluripotent stem cell sources-potentially useful for the development of new therapeutic approaches. Starting with skin fibroblasts from patients diagnosed with cystic fibrosis, we derived and characterized induced pluripotent stem cell (iPSC) lines.We then utilized zinc-finger nucleases (ZFNs), designed to target the endogenous CFTR gene, to mediate correction of the inherited genetic mutation in these patientderived lines via homology-directed repair (HDR). We observed an exquisitely sensitive, homology-dependent preference for targeting one CFTR allele versus the other. The corrected cystic fibrosis iPSCs, when induced to differentiate in vitro, expre{\ss}ed the corrected CFTR gene; importantly, CFTR correction resulted in restored expre{\ss}ion of the mature CFTR glycoprotein and restoration of CFTR chloride channel function in iPSC-derived epithelial cells.",

author = "Crane, \{Ana M.\} and Philipp Kramer and Bui, \{Jacquelin H.\} and Chung, \{Wook Joon\} and Li, \{Xuan Shirley\} and Gonzalez-Garay, \{Manuel L.\} and Finn Hawkins and Wei Liao and Daniela Mora and Sangbum Choi and Jianbin Wang and Sun, \{Helena C.\} and Paschon, \{David E.\} and Guschin, \{Dmitry Y.\} and Gregory, \{Philip D.\} and Kotton, \{Darrell N.\} and Holmes, \{Michael C.\} and Sorscher, \{Eric J.\} and Davis, \{Brian R.\}",

note = "Funding Information: We thank Naoki Nakayama (Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center) for helpful discussions; Veronica K. Quiceno for editorial assistance; Kathryn Plath, Connie Cepko, Brian Sauer, and Jeffrey Beekman for providing DNA constructs either directly or through Addgene; and Fyodor Urnov (Sangamo BioSciences, Inc.) for ZFN development through funding from the Cystic Fibrosis Foundation (CFF) Folding Consortium. This study was supported by NIH RC1HL099559 (B.R.D. and R. Wetsel, primary investigators [PIs]); CFF DAVIS12GO (B.R.D., PI); NIH P30 DK072482 (E.J.S., PI); and CFF R464 (E.J.S., PI). J.W., H.C.S., D.E.P., D.Y.G, P.D.G., and M.C.H. are employees of Sangamo BioSciences, Inc. ",

year = "2015",

doi = "10.1016/j.stemcr.2015.02.005",

language = "English",

volume = "4",

pages = "569--577",

journal = "Stem Cell Reports",

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T1 - Targeted correction and restored function of the CFTR gene in cystic fibrosis induced pluripotent stem cells

AU - Crane, Ana M.

AU - Kramer, Philipp

AU - Bui, Jacquelin H.

AU - Chung, Wook Joon

AU - Li, Xuan Shirley

AU - Gonzalez-Garay, Manuel L.

AU - Hawkins, Finn

AU - Liao, Wei

AU - Mora, Daniela

AU - Choi, Sangbum

AU - Wang, Jianbin

AU - Sun, Helena C.

AU - Paschon, David E.

AU - Guschin, Dmitry Y.

AU - Gregory, Philip D.

AU - Kotton, Darrell N.

AU - Holmes, Michael C.

AU - Sorscher, Eric J.

AU - Davis, Brian R.

N1 - Funding Information: We thank Naoki Nakayama (Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center) for helpful discussions; Veronica K. Quiceno for editorial assistance; Kathryn Plath, Connie Cepko, Brian Sauer, and Jeffrey Beekman for providing DNA constructs either directly or through Addgene; and Fyodor Urnov (Sangamo BioSciences, Inc.) for ZFN development through funding from the Cystic Fibrosis Foundation (CFF) Folding Consortium. This study was supported by NIH RC1HL099559 (B.R.D. and R. Wetsel, primary investigators [PIs]); CFF DAVIS12GO (B.R.D., PI); NIH P30 DK072482 (E.J.S., PI); and CFF R464 (E.J.S., PI). J.W., H.C.S., D.E.P., D.Y.G, P.D.G., and M.C.H. are employees of Sangamo BioSciences, Inc.

PY - 2015

Y1 - 2015

N2 - Recently developed reprogramming and genome editing technologies make poßible the derivation of corrected patient-specific pluripotent stem cell sources-potentially useful for the development of new therapeutic approaches. Starting with skin fibroblasts from patients diagnosed with cystic fibrosis, we derived and characterized induced pluripotent stem cell (iPSC) lines.We then utilized zinc-finger nucleases (ZFNs), designed to target the endogenous CFTR gene, to mediate correction of the inherited genetic mutation in these patientderived lines via homology-directed repair (HDR). We observed an exquisitely sensitive, homology-dependent preference for targeting one CFTR allele versus the other. The corrected cystic fibrosis iPSCs, when induced to differentiate in vitro, expreßed the corrected CFTR gene; importantly, CFTR correction resulted in restored expreßion of the mature CFTR glycoprotein and restoration of CFTR chloride channel function in iPSC-derived epithelial cells.

AB - Recently developed reprogramming and genome editing technologies make poßible the derivation of corrected patient-specific pluripotent stem cell sources-potentially useful for the development of new therapeutic approaches. Starting with skin fibroblasts from patients diagnosed with cystic fibrosis, we derived and characterized induced pluripotent stem cell (iPSC) lines.We then utilized zinc-finger nucleases (ZFNs), designed to target the endogenous CFTR gene, to mediate correction of the inherited genetic mutation in these patientderived lines via homology-directed repair (HDR). We observed an exquisitely sensitive, homology-dependent preference for targeting one CFTR allele versus the other. The corrected cystic fibrosis iPSCs, when induced to differentiate in vitro, expreßed the corrected CFTR gene; importantly, CFTR correction resulted in restored expreßion of the mature CFTR glycoprotein and restoration of CFTR chloride channel function in iPSC-derived epithelial cells.

UR - https://www.scopus.com/pages/publications/84933677842

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DO - 10.1016/j.stemcr.2015.02.005

M3 - Article

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AN - SCOPUS:84933677842

SN - 2213-6711

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EP - 577

JO - Stem Cell Reports

JF - Stem Cell Reports

IS - 4

ER -

Targeted correction and restored function of the CFTR gene in cystic fibrosis induced pluripotent stem cells

Abstract

Bibliographical note

ASJC Scopus subject areas

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