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Cell Rep. 2017 Jul 11;20(2):384-396. doi: 10.1016/j.celrep.2017.06.045.

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

Author information

1
Neurobiology-Neurodegeneration and Repair Laboratory, National Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA.
2
Laboratory of Cell and Developmental Signaling, National Cancer Institute - Frederick, Frederick, MD 21702, USA.
3
Electron Microscope Laboratory, Leidos Biomedical Research, Inc., National Cancer Institute - Frederick, Frederick, MD 21702, USA.
4
Department of Ophthalmology, Scheie Eye Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
5
Pediatric, Developmental, and Genetic Eye Disease Branch, National Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA.
6
National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA; Department of Pediatrics, McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
7
Laboratory of Cell and Developmental Signaling, National Cancer Institute - Frederick, Frederick, MD 21702, USA. Electronic address: chris.westlake@nih.gov.
8
Neurobiology-Neurodegeneration and Repair Laboratory, National Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA. Electronic address: swaroopa@nei.nih.gov.

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

PMID:
28700940
PMCID:
PMC5553702
DOI:
10.1016/j.celrep.2017.06.045
[Indexed for MEDLINE]
Free PMC Article

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