CRISPR-based gene editing enables FOXP3 gene repair in IPEX patient cells

M. Goodwin; E. Lee; U. Lakshmanan; S. Shipp; L. Froessl; F. Barzaghi; L. Passerini; M. Narula; A. Sheikali; C. M. Lee; G. Bao; C. S. Bauer; H. K. Miller; M. Garcia-Lloret; M. J. Butte; A. Bertaina; A. Shah; M. Pavel-Dinu; A. Hendel; M. Porteus; M. G. Roncarolo; R. Bacchetta

doi:10.1126/sciadv.aaz0571

CRISPR-based gene editing enables FOXP3 gene repair in IPEX patient cells

M. Goodwin, E. Lee, U. Lakshmanan, S. Shipp, L. Froessl, F. Barzaghi, L. Passerini, M. Narula, A. Sheikali, C. M. Lee, G. Bao, C. S. Bauer, H. K. Miller, M. Garcia-Lloret, M. J. Butte, A. Bertaina, A. Shah, M. Pavel-Dinu, A. Hendel, M. PorteusM. G. Roncarolo, R. Bacchetta

The Mina and Everard Goodman Faculty of Life Sciences

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

84 Scopus citations

Abstract

The prototypical genetic autoimmune disease is immune dysregulation polyendocrinopathy enteropathy X-linked (IPEX) syndrome, a severe pediatric disease with limited treatment options. IPEX syndrome is caused by mutations in the forkhead box protein 3 (FOXP3) gene, which plays a critical role in immune regulation. As a monogenic disease, IPEX is an ideal candidate for a therapeutic approach in which autologous hematopoietic stem and progenitor (HSPC) cells or T cells are gene edited ex vivo and reinfused. Here, we describe a CRISPR-based gene correction permitting regulated expression of FOXP3 protein. We demonstrate that gene editing preserves HSPC differentiation potential, and that edited regulatory and effector T cells maintain their in vitro phenotype and function. Additionally, we show that this strategy is suitable for IPEX patient cells with diverse mutations. These results demonstrate the feasibility of gene correction, which will be instrumental for the development of therapeutic approaches for other genetic autoimmune diseases.

Original language	English
Article number	eaaz0571
Journal	Science advances
Volume	6
Issue number	19
DOIs	https://doi.org/10.1126/sciadv.aaz0571
State	Published - May 2020

Bibliographical note

Publisher Copyright:
Copyright © 2020 The Authors,

Funding

We gratefully acknowledge the support from several grants and awards, including the NIH NIAID (R21 AI123896), the CIRM (DISC2-09526), the Stanford SPARK (1110504-308-DHBTC), and the Sutardja Foundation (1198779-101-GHFBB), and the Cancer Prevention and Research Institute of Texas to G.B. (RR140081 and RR170721), in addition to a donation from an anonymous donor to the Stanford Center for Genetic Immune Diseases (CGID) for immunological studies on the patient samples. M.G. thanks the Stanford University for the tremendous support, including the Stanford Maternal and Child Health Research Institute (MCHRI) Postdoctoral Award (Stanford NIH-NCATS-CTSA, UL1 TR001085) and the Stanford University Office of the Dean Postdoctoral Award (1182084-100-DHDEZ). E.L. received funding support from the Agency for Science, Technology and Research, Singapore and the Stanford Masters of Science in Medicine scholarship.

Funders	Funder number
Agency for Science, Technology and Research, Singapore
MCHRI
NIH NIAID	R21 AI123896
Stanford Masters of Science in Medicine
Stanford Maternal and Child Health Research Institute
Stanford NIH-NCATS-CTSA	UL1 TR001085, 1182084-100-DHDEZ
Sutardja Foundation	1198779-101-GHFBB
California Institute for Regenerative Medicine	DISC2-09526, 1110504-308-DHBTC
Cancer Prevention and Research Institute of Texas	RR170721, RR140081
Stanford University

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Goodwin, M., Lee, E., Lakshmanan, U., Shipp, S., Froessl, L., Barzaghi, F., Passerini, L., Narula, M., Sheikali, A., Lee, C. M., Bao, G., Bauer, C. S., Miller, H. K., Garcia-Lloret, M., Butte, M. J., Bertaina, A., Shah, A., Pavel-Dinu, M., Hendel, A., ... Bacchetta, R. (2020). CRISPR-based gene editing enables FOXP3 gene repair in IPEX patient cells. Science advances, 6(19), Article eaaz0571. https://doi.org/10.1126/sciadv.aaz0571

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title = "CRISPR-based gene editing enables FOXP3 gene repair in IPEX patient cells",

abstract = "The prototypical genetic autoimmune disease is immune dysregulation polyendocrinopathy enteropathy X-linked (IPEX) syndrome, a severe pediatric disease with limited treatment options. IPEX syndrome is caused by mutations in the forkhead box protein 3 (FOXP3) gene, which plays a critical role in immune regulation. As a monogenic disease, IPEX is an ideal candidate for a therapeutic approach in which autologous hematopoietic stem and progenitor (HSPC) cells or T cells are gene edited ex vivo and reinfused. Here, we describe a CRISPR-based gene correction permitting regulated expression of FOXP3 protein. We demonstrate that gene editing preserves HSPC differentiation potential, and that edited regulatory and effector T cells maintain their in vitro phenotype and function. Additionally, we show that this strategy is suitable for IPEX patient cells with diverse mutations. These results demonstrate the feasibility of gene correction, which will be instrumental for the development of therapeutic approaches for other genetic autoimmune diseases.",

author = "M. Goodwin and E. Lee and U. Lakshmanan and S. Shipp and L. Froessl and F. Barzaghi and L. Passerini and M. Narula and A. Sheikali and Lee, {C. M.} and G. Bao and Bauer, {C. S.} and Miller, {H. K.} and M. Garcia-Lloret and Butte, {M. J.} and A. Bertaina and A. Shah and M. Pavel-Dinu and A. Hendel and M. Porteus and Roncarolo, {M. G.} and R. Bacchetta",

note = "Publisher Copyright: Copyright {\textcopyright} 2020 The Authors,",

year = "2020",

month = may,

doi = "10.1126/sciadv.aaz0571",

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Goodwin, M, Lee, E, Lakshmanan, U, Shipp, S, Froessl, L, Barzaghi, F, Passerini, L, Narula, M, Sheikali, A, Lee, CM, Bao, G, Bauer, CS, Miller, HK, Garcia-Lloret, M, Butte, MJ, Bertaina, A, Shah, A, Pavel-Dinu, M, Hendel, A, Porteus, M, Roncarolo, MG & Bacchetta, R 2020, 'CRISPR-based gene editing enables FOXP3 gene repair in IPEX patient cells', Science advances, vol. 6, no. 19, eaaz0571. https://doi.org/10.1126/sciadv.aaz0571

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AU - Goodwin, M.

AU - Lee, E.

AU - Lakshmanan, U.

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AU - Barzaghi, F.

AU - Passerini, L.

AU - Narula, M.

AU - Sheikali, A.

AU - Lee, C. M.

AU - Bao, G.

AU - Bauer, C. S.

AU - Miller, H. K.

AU - Garcia-Lloret, M.

AU - Butte, M. J.

AU - Bertaina, A.

AU - Shah, A.

AU - Pavel-Dinu, M.

AU - Hendel, A.

AU - Porteus, M.

AU - Roncarolo, M. G.

AU - Bacchetta, R.

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CRISPR-based gene editing enables FOXP3 gene repair in IPEX patient cells

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