@article{9aacb87fc5bd49fd9f64ce1f9518caca,
title = "In Vitro Modeling Using Ciliopathy-Patient-Derived Cells Reveals Distinct Cilia Dysfunctions Caused by CEP290 Mutations",
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.",
keywords = "Hedgehog signaling, Joubert syndrome, LCA, ciliary transition zone, ciliogenesis, ciliopathies, iPSC, organoid, primary cilia, retinal degeneration",
author = "Hiroko Shimada and Quanlong Lu and Christine Insinna-Kettenhofen and Kunio Nagashima and English, {Milton A.} and Semler, {Elizabeth M.} and Jacklyn Mahgerefteh and Cideciyan, {Artur V.} and Tiansen Li and Brooks, {Brian P.} and Meral Gunay-Aygun and Jacobson, {Samuel G.} and Tiziana Cogliati and Westlake, {Christopher J.} and Anand Swaroop",
note = "Funding Information: We are grateful to the patients and families who participated in this investigation. We sincerely thank Mahendra Rao for advice, Guokai Chen and Jeanette Beers for generating iPSC lines, Chengyu Liu for teratoma formation, Rossukon Kaewkhaw and May C. Malicdan for isolation of fibroblasts from skin biopsy specimens, Robert Fariss and Chun Gao for assistance with microscopy, and Pradeep Kota for help with FRAP analysis. We acknowledge N-NRL colleagues Lauren Bouchard Killingsworth, Yu Holly Chen, Yogita Adlakha, Mugdha Samant, and Jacob Nellissery for assistance in cell culture experiments. This research was supported by the Intramural Research Program of the National Eye Institute (grants EY000450 and EY000546) and the National Human Genome Research Institute, the Japan Society?for the Promotion of Science (JSPS grant #71411), the Joubert Syndrome and Related Disorders Foundation, and National Cancer Institute contract HHSN26120080001E. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the US Government. Publisher Copyright: {\textcopyright} 2017",
year = "2017",
month = jul,
day = "11",
doi = "10.1016/j.celrep.2017.06.045",
language = "English (US)",
volume = "20",
pages = "384--396",
journal = "Cell Reports",
issn = "2211-1247",
publisher = "Cell Press",
number = "2",
}