Disruption of CEP290 microtubule/membrane-binding domains causes retinal degeneration

Abstract

Mutations in the gene centrosomal protein 290 kDa (CEP290) cause an array of debilitating and phenotypically distinct human diseases, ranging from the devastating blinding disease Leber congenital amaurosis (LCA) to Senior-Løken syndrome, Joubert syndrome, and the lethal Meckel-Gruber syndrome. Despite its critical role in biology and disease, very little is known about CEP290's function. Here, we have identified 4 functional domains of the protein. We found that CEP290 directly binds to cellular membranes through an N-terminal domain that includes a highly conserved amphipathic helix motif and to microtubules through a domain located within its myosin-tail homology domain. Furthermore, CEP290 activity was regulated by 2 autoinhibitory domains within its N and C termini, both of which were found to play critical roles in regulating ciliogenesis. Disruption of the microtubule-binding domain in a mouse model of LCA was sufficient to induce significant deficits in cilium formation, which led to retinal degeneration. These data implicate CEP290 as an integral structural and regulatory component of the cilium and provide insight into the pathological mechanisms of LCA and related ciliopathies. Further, these data illustrate that disruption of particular CEP290 functional domains may lead to particular disease phenotypes and suggest innovative strategies for therapeutic intervention.

Clinical Characteristics and Current Therapies for Inherited Retinal Degenerations

“…Intriguingly, mutations in CEP290 have been described in 50% of Jou-bert syndrome subgroup of ciliopathies and up to 20% of cases of Leber congenital amaurosis, and found to be associated with a wide variety of distinct phenotypes, including Senior-Løken syndrome, nephronophthisis, Bardet-Biedl syndrome, and Meckel-Gruber syndrome (for review Coppieters et al 2010;Waters and Beales 2011;Drivas et al 2013). At present, despite the identification of more than 100 CEP290 mutations (Coppieters et al 2010), no clear genotype -phenotype correlations could be established and the mechanisms underlying the highly variable phenotypes associated with ciliary dysfunction remain to be elucidated.…”

The Status ofRPE65Gene Therapy Trials: Safety and Efficacy

“…Other viral or nonviral vectors may also be useful for those conditions (Kong et al 2008;Puppo et al 2014). Alternative strategies, such as use of a truncated cDNA, trans-splicing, gene correction, or delivery of antisense nucleic acids may be effective (Lai et al 2008;Collin et al 2012;Drivas et al 2013).…”

Patient-specific induced pluripotent stem cells (iPSCs) for the study and treatment of retinal degenerative diseases

“…LCA is predominantly inherited in an autosomal recessive manner and of the seventeen loci associated with the disease, the largest contributor is CEP290 (Perrault et al, 2007;Stone, 2007). Thirty percent of LCA patients carry mutations in CEP290, the gene product of which is a cilia-associated protein that functions in photoreceptor outer segment trafficking and ciliogenesis (Craige et al, 2010;Drivas et al, 2013;McEwen et al, 2007;Tsang et al, 2008). Loss of photoreceptor outer segments in patients with CEP290-associated LCA results in severe vision loss.…”