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

• Published in: Cell reports (Cambridge) 2017-07, Vol.20 (2), p.384-396
• Main Authors: Shimada, Hiroko, Lu, Quanlong, Insinna-Kettenhofen, Christine, Nagashima, Kunio, English, Milton A., Semler, Elizabeth M., Mahgerefteh, Jacklyn, Cideciyan, Artur V., Li, Tiansen, Brooks, Brian P., Gunay-Aygun, Meral, Jacobson, Samuel G., Cogliati, Tiziana, Westlake, Christopher J., Swaroop, Anand
• Format: Article
• Language: English
• Subjects: Adenylyl Cyclases - genetics; Adenylyl Cyclases - metabolism; ADP-Ribosylation Factors - genetics; ADP-Ribosylation Factors - metabolism; Alleles; Animals; Antigens, Neoplasm - genetics; Antigens, Neoplasm - metabolism; Cilia; ciliary transition zone; ciliogenesis; ciliopathies; Fibroblasts - metabolism; Hedgehog Proteins - genetics; Hedgehog Proteins - metabolism; Hedgehog signaling; Homozygote; Humans; iPSC; Joubert syndrome; LCA; Mice; Mice, Knockout; Mutation - genetics; Neoplasm Proteins - genetics; Neoplasm Proteins - metabolism; organoid; primary cilia; Receptors, G-Protein-Coupled - genetics; Receptors, G-Protein-Coupled - metabolism; retinal degeneration; Smoothened Receptor - genetics; Smoothened Receptor - metabolism
• ISSN: 2211-1247; 2211-1247
• DOI: 10.1016/j.celrep.2017.06.045

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. [Display omitted] •Normal cilia in fibroblasts, but not in photoreceptors of CEP290-LCA organoids•No CEP290 protein and defective ciliogenesis in CEP290-JSRD fibroblasts•Selective reduction in ciliary localization of ARL13B and ADCY3 in JSRD fibroblasts•Cilia defects in JSRD fibroblasts affect Smo and GPR161 transport in Hh signaling Using fibroblasts and iPSC-derived optic cups from patients with distinct CEP290 mutations, Shimada et al. show a concordance between ciliogenesis defects in different cell types and clinical severity in Leber congenital amaurosis and Joubert syndrome. These studies establish CEP290 as gatekeeper of signaling molecules in and out of the primary cilium.