CEP290 gene transfer rescues Leber congenital amaurosis cellular phenotype

Abstract

Mutations in CEP290 are the most common cause of Leber congenital amaurosis (LCA), a severe inherited retinal degenerative disease for which there is currently no cure. Autosomal recessive CEP290-associated LCA is a good candidate for gene-replacement therapy, and cells derived from affected individuals give researchers the ability to study human disease and therapeutic gene correction in vitro. Here we report the development of lentiviral vectors carrying full-length CEP290 for the purpose of correcting the CEP290 disease-specific phenotype in human cells. A lentiviral vector containing CMV-driven human full-length CEP290 was constructed. Following transduction of patient-specific, iPSC-derived, photoreceptor precursor cells, rt-PCR analysis and western blotting revealed vector-derived expression. Because CEP290 is important in ciliogenesis, the ability of fibroblast cultures from CEP290-associated LCA patients to form cilia was investigated. In cultures derived from these patients, fewer cells formed cilia compared to unaffected controls. Cilia that were formed were shorter in patient derived cells than in cells from unaffected individuals. Importantly, lentiviral delivery of CEP290 rescued the ciliogenesis defect. The successful construction and viral transfer of full-length CEP290 brings us closer to the goal of providing gene- and cell- based therapies for patients affected with this common form of LCA.

Eupatilin rescues ciliary transition zone defects to ameliorate ciliopathy-related phenotypes

“…CEP290 is a gene frequently mutated in several ciliopathies and LCA (6). CEP290 gene augmentation therapy using viral vectors has been shown to ameliorate LCA-related phenotypes in human cell and mouse models (7,8). Oligonucleotide-mediated splicing correction has also been shown to rescue LCA phenotypes in the induced pluripotent stem cell-derived optic cups of patients harboring a CEP290 intronic mutation (9).…”

Mixing-sequence-dependent nucleic acid complexation and gene transfer efficiency by polyethylenimine

“…[1][2][3][4] Gene therapy started from a concept of correcting an abnormal gene by delivering a desirable therapeutic transgene (i.e., DNA), 5 and has recently evolved to interfere with target biological activities at a translational level by delivering small nucleic acids (e.g., siRNA and antisense ODN). 6,7 In order to address the clinical challenges associated with using viral vectors (e.g., immunogenicity, onco/tumorigenicity, and inefficient and cumbersome preparation), 8,9 developing nonviral vectors that are as efficient as viral counterparts with improved safety measures has been of great interest.…”

Induced Pluripotent Stem Cell Therapies for Degenerative Disease of the Outer Retina: Disease Modeling and Cell Replacement

“…61 iPSC-derived photoreceptor precursor cells carrying an USH2A mutation showed increased ER stress due to misfolding of the protein. 61 iPSCderived photoreceptor precursors, this time from a patient with Leber congenital amaurosis caused by mutation in CEP290, have been used to test lentiviral vectors for gene replacement therapy 62 ; viral transduction improved the ciliogenesis defect in cells from affected individuals.…”