In Vitro Modeling Using Ciliopathy-Patient-Derived Cells Reveals Distinct Cilia Dysfunctions Caused by CEP290 Mutations

Hiroko Shimada; Quanlong Lu; Christine Insinna-Kettenhofen; Kunio Nagashima; Milton A. English; Elizabeth M. Semler; Jacklyn Mahgerefteh; Artur V. Cideciyan; Tiansen Li; Brian P. Brooks; Meral Gunay-Aygun; Samuel G. Jacobson; Tiziana Cogliati; Christopher J. Westlake; Anand Swaroop

doi:10.1016/j.celrep.2017.06.045

In Vitro Modeling Using Ciliopathy-Patient-Derived Cells Reveals Distinct Cilia Dysfunctions Caused by CEP290 Mutations

Hiroko Shimada, Quanlong Lu, Christine Insinna-Kettenhofen, Kunio Nagashima, Milton A. English, Elizabeth M. Semler, Jacklyn Mahgerefteh, Artur V. Cideciyan, Tiansen Li, Brian P. Brooks, Meral Gunay-Aygun, Samuel G. Jacobson, Tiziana Cogliati, Christopher J. Westlake, Anand Swaroop

School of Medicine

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

58 Scopus citations

Abstract

Mutations in CEP290, a transition zone protein in primary cilia, cause diverse ciliopathies, including Leber congenital amaurosis (LCA) and Joubert-syndrome and related disorders (JSRD). We examined cilia biogenesis and function in cells derived from CEP290-LCA and CEP290-JSRD patients. CEP290 protein was reduced in LCA fibroblasts with no detectable impact on cilia; however, optic cups derived from induced pluripotent stem cells (iPSCs) of CEP290-LCA patients displayed less developed photoreceptor cilia. Lack of CEP290 in JSRD fibroblasts resulted in abnormal cilia and decreased ciliogenesis. We observed selectively reduced localization of ADCY3 and ARL13B. Notably, Hedgehog signaling was augmented in CEP290-JSRD because of enhanced ciliary transport of Smoothened and GPR161. These results demonstrate a direct correlation between the extent of ciliogenesis defects in fibroblasts and photoreceptors with phenotypic severity in JSRD and LCA, respectively, and strengthen the role of CEP290 as a selective ciliary gatekeeper for transport of signaling molecules in and out of the cilium.

Original language	English (US)
Pages (from-to)	384-396
Number of pages	13
Journal	Cell Reports
Volume	20
Issue number	2
DOIs	https://doi.org/10.1016/j.celrep.2017.06.045
State	Published - Jul 11 2017

Keywords

Hedgehog signaling
Joubert syndrome
LCA
ciliary transition zone
ciliogenesis
ciliopathies
iPSC
organoid
primary cilia
retinal degeneration

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology

Access to Document

10.1016/j.celrep.2017.06.045

Cite this

Shimada, H., Lu, Q., Insinna-Kettenhofen, C., Nagashima, K., English, M. A., Semler, E. M., Mahgerefteh, J., Cideciyan, A. V., Li, T., Brooks, B. P., Gunay-Aygun, M., Jacobson, S. G., Cogliati, T., Westlake, C. J., & Swaroop, A. (2017). In Vitro Modeling Using Ciliopathy-Patient-Derived Cells Reveals Distinct Cilia Dysfunctions Caused by CEP290 Mutations. Cell Reports, 20(2), 384-396. https://doi.org/10.1016/j.celrep.2017.06.045

Shimada, H, Lu, Q, Insinna-Kettenhofen, C, Nagashima, K, English, MA, Semler, EM, Mahgerefteh, J, Cideciyan, AV, Li, T, Brooks, BP, Gunay-Aygun, M, Jacobson, SG, Cogliati, T, Westlake, CJ & Swaroop, A 2017, 'In Vitro Modeling Using Ciliopathy-Patient-Derived Cells Reveals Distinct Cilia Dysfunctions Caused by CEP290 Mutations', Cell Reports, vol. 20, no. 2, pp. 384-396. https://doi.org/10.1016/j.celrep.2017.06.045

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abstract = "Mutations in CEP290, a transition zone protein in primary cilia, cause diverse ciliopathies, including Leber congenital amaurosis (LCA) and Joubert-syndrome and related disorders (JSRD). We examined cilia biogenesis and function in cells derived from CEP290-LCA and CEP290-JSRD patients. CEP290 protein was reduced in LCA fibroblasts with no detectable impact on cilia; however, optic cups derived from induced pluripotent stem cells (iPSCs) of CEP290-LCA patients displayed less developed photoreceptor cilia. Lack of CEP290 in JSRD fibroblasts resulted in abnormal cilia and decreased ciliogenesis. We observed selectively reduced localization of ADCY3 and ARL13B. Notably, Hedgehog signaling was augmented in CEP290-JSRD because of enhanced ciliary transport of Smoothened and GPR161. These results demonstrate a direct correlation between the extent of ciliogenesis defects in fibroblasts and photoreceptors with phenotypic severity in JSRD and LCA, respectively, and strengthen the role of CEP290 as a selective ciliary gatekeeper for transport of signaling molecules in and out of the cilium.",

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AU - Shimada, Hiroko

AU - Lu, Quanlong

AU - Insinna-Kettenhofen, Christine

AU - Nagashima, Kunio

AU - English, Milton A.

AU - Semler, Elizabeth M.

AU - Mahgerefteh, Jacklyn

AU - Cideciyan, Artur V.

AU - Li, Tiansen

AU - Brooks, Brian P.

AU - Gunay-Aygun, Meral

AU - Jacobson, Samuel G.

AU - Cogliati, Tiziana

AU - Westlake, Christopher J.

AU - Swaroop, Anand

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AB - Mutations in CEP290, a transition zone protein in primary cilia, cause diverse ciliopathies, including Leber congenital amaurosis (LCA) and Joubert-syndrome and related disorders (JSRD). We examined cilia biogenesis and function in cells derived from CEP290-LCA and CEP290-JSRD patients. CEP290 protein was reduced in LCA fibroblasts with no detectable impact on cilia; however, optic cups derived from induced pluripotent stem cells (iPSCs) of CEP290-LCA patients displayed less developed photoreceptor cilia. Lack of CEP290 in JSRD fibroblasts resulted in abnormal cilia and decreased ciliogenesis. We observed selectively reduced localization of ADCY3 and ARL13B. Notably, Hedgehog signaling was augmented in CEP290-JSRD because of enhanced ciliary transport of Smoothened and GPR161. These results demonstrate a direct correlation between the extent of ciliogenesis defects in fibroblasts and photoreceptors with phenotypic severity in JSRD and LCA, respectively, and strengthen the role of CEP290 as a selective ciliary gatekeeper for transport of signaling molecules in and out of the cilium.

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