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.

• Authors: 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
• Journal: Cell reports (Cell Rep) Vol. 20 Issue 2 Pg. 384-396 (07 11 2017) ISSN: 2211-1247 [Electronic] United States
• PMID: 28700940 (Publication Type: Journal Article)
• Copyright: Published by Elsevier Inc.

Chemical References

• Antigens, Neoplasm
• Arl13b protein, mouse
• Cell Cycle Proteins
• Cep290 protein, human
• Cytoskeletal Proteins
• GPR161 protein, mouse
• Hedgehog Proteins
• Neoplasm Proteins
• Receptors, G-Protein-Coupled
• Smo protein, mouse
• Smoothened Receptor
• ADP-Ribosylation Factors
• Adenylyl Cyclases
• adenylate cyclase 3

Topics

• ADP-Ribosylation Factors (genetics, metabolism)
• Adenylyl Cyclases (genetics, metabolism)
• Alleles
• Animals
• Antigens, Neoplasm (genetics, metabolism)
• Cell Cycle Proteins
• Cilia
• Cytoskeletal Proteins
• Fibroblasts (metabolism)
• Hedgehog Proteins (genetics, metabolism)
• Homozygote
• Humans
• Mice
• Mice, Knockout
• Mutation (genetics)
• Neoplasm Proteins (genetics, metabolism)
• Receptors, G-Protein-Coupled (genetics, metabolism)
• Smoothened Receptor (genetics, metabolism)