Molecular cloning and expression of RPE65, a novel retinal pigment epithelium-specific microsomal protein that is post-transcriptionally regulated in vitro

Hamel, Christian; Tsilou, Ekaterini; Pfeffer, Bruce A.; Hooks, John J.; Detrick, Barbara; Redmond, T. Michael

doi:10.1016/s0021-9258(18)82319-5

Cited by 287 publications

(52 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…It is estimated that there are over 2.5 million individuals affected by IRDs worldwide [25], with RP being the most common form. The prevalence of RP is approximately 1:4000 [3,26,27]. In particular, between 21% and 54% of IRD patients have RP [28,29].…”

Section: Epidemiology Of Rpe65-irdsmentioning

confidence: 99%

Inherited Retinal Diseases Due to RPE65 Variants: From Genetic Diagnostic Management to Therapy

Aoun

Passerini

Chiurazzi

et al. 2021

IJMS

View full text Add to dashboard Cite

Inherited retinal diseases (IRDs) are a heterogeneous group of conditions that include retinitis pigmentosa (RP) and Leber congenital amaurosis (LCA) and early-onset severe retinal dystrophy (EO[S]RD), which differ in severity and age of onset. IRDs are caused by mutations in >250 genes. Variants in the RPE65 gene account for 0.6–6% of RP and 3–16% of LCA/EORD cases. Voretigene neparvovec is a gene therapy approved for the treatment of patients with an autosomal recessive retinal dystrophy due to confirmed biallelic RPE65 variants (RPE65-IRDs). Therefore, the accurate molecular diagnosis of RPE65-IRDs is crucial to identify ‘actionable’ genotypes—i.e., genotypes that may benefit from the treatment—and is an integral part of patient management. To date, hundreds of RPE65 variants have been identified, some of which are classified as pathogenic or likely pathogenic, while the significance of others is yet to be established. In this review, we provide an overview of the genetic diagnostic workup needed to select patients that could be eligible for voretigene neparvovec treatment. Careful clinical characterization of patients by multidisciplinary teams of experts, combined with the availability of next-generation sequencing approaches, can accelerate patients’ access to available therapeutic options.

show abstract

Section: Epidemiology Of Rpe65-irdsmentioning

confidence: 99%

Inherited Retinal Diseases Due to RPE65 Variants: From Genetic Diagnostic Management to Therapy

Aoun

Passerini

Chiurazzi

et al. 2021

IJMS

View full text Add to dashboard Cite

show abstract

“…Nevertheless, RPE65 has been reported to be membrane associated but not to be an integral membrane protein because it is lacking hydrophobic transmembrane domains and a signal peptide (Bavik et al, 1993;Hamel et al, 1993;Nicoletti et al, 1995). The knowledge on the structure of the protein is limited, and the possible receptor function of RPE65 for plasma RBP has been discussed and is still unclear (Bavik et al, 1993;Hamel et al, 1993;Nicoletti et al, 1995;Redmond et al, 1998;Flower, 2000). As abundantly expressed in retinal pigment epithelial cells, RPE65 was shown to be involved in the all-trans-to 11-cis-isomerization reaction that regenerates the 11-cis-retinal chromophore of rhodopsin in the visual cycle of the retina (Redmond et al, 1998).…”

Section: Figurementioning

confidence: 99%

“…Some authors show data supporting the hypothesis of receptor independent delivery of retinol to human keratinocytes (Hodam et al, 1991;Hodam and Creek, 1998), and, in contrast, studies propose that a regulated retinol transfer into human keratinocytes implies involvement of a receptor protein to which the RBP of the RBP-retinol complex is bound (Bavik et al, 1995;Smeland et al, 1995). A putative receptor for plasma RBP has been assigned to a 61-kDa protein (named RPE65) which was originally identified in retinal pigment epithelium cells (Bavik et al, 1992;Hamel et al, 1993;Bavik et al, 1995;Nicoletti et al, 1995). This protein is highly conserved across various species and is predominantly expressed in the retinal pigment epithelium (Bavik et al, 1993;Nicoletti et al, 1995).…”

mentioning

confidence: 99%

“…The physiologic role of RPE65 is under debate. It has been reported to be membrane associated but not to be an integral membrane protein lacking hydrophobic transmembrane domains and signaling peptides (Bavik et al, 1992;Hamel et al, 1993;Nicoletti et al, 1995;Flower, 2000). The abundant expression in retinal pigment epithelium indicates an important role for RPE65 in retinoid processing and transport in the eye suggesting to be involved in human retinal degenerations (Nicoletti et al, 1995).…”

mentioning

confidence: 99%

See 1 more Smart Citation

RPE65 of Retinal Pigment Epithelium, A Putative Receptor Molecule for Plasma Retinol-Binding Protein, is Expressed in Human Keratinocytes

Hinterhuber¹,

Cauza²,

Brugger³

et al. 2004

Journal of Investigative Dermatology

View full text Add to dashboard Cite

Retinoids are important modulators for cell growth and differentiation of normal skin. In plasma, retinol is transported coupled to plasma retinol-binding protein. In this study, we investigated gene and protein expression of RPE65, a putative receptor for plasma retinol-binding protein in human epidermal keratinocytes. We performed real-time PCR analysis to evaluate expression of RPE65 mRNA in proliferating and differentiating keratinocytes. Immunoblotting with anti-RPE65 antibody shows distinct reactivity to a 61-kDa protein. Indirect immunofluorescence on normal human epidermis reveals cell surface labeling of keratinocytes. Laser scan microscopy exhibits colocalization of plasma retinol-binding protein and RPE65 on cultured keratinocytes. Internalization experiments with [3H]retinoic acid-retinol-binding protein complex in the presence and absence of excess of retinol-binding protein indicates receptor-dependent uptake of retinoids. We further show isolation of RPE65 protein by affinity chromatography from lysates of keratinocytes using a retinol-binding protein-matrix gel column. In summary, we demonstrate mRNA and protein expression of RPE65 in epidermal keratinocytes. Colocalization of plasma retinol-binding protein with RPE65 and affinity binding suggest a direct interaction of RPE65 with plasma retinol-binding protein in cultured human keratinocytes that might be involved in retinoid uptake of keratinocytes.

show abstract

“…Photons of light, captured in the photoreceptor outer segment, isomerise 11-cis-retinal to all-trans-retinal, which then requires recycling to restore 11-cis-retinal. RPE65, the most well-known enzyme in this pathway, catalyses one critical step in regenerating 11-cis-retinal in the RPE, [1][2][3] and loss of enzymatic function is associated with an early-onset rod-cone dystrophy (Leber congenital amaurosis, LCA type 2). [4][5][6] RPE65 gene replacement therapy is now available to treat this.…”

Section: Introductionmentioning

confidence: 99%

Novel disease-causing variant in RDH12 presenting with autosomal dominant retinitis pigmentosa

et al. 2021

View full text Add to dashboard Cite

AimTo describe the clinical and molecular features of a novel, autosomal dominant RDH12-retinopathy.MethodsRetrospective cross-sectional study. Twelve individuals from a four-generation British pedigree underwent ophthalmic examination, genotyping using next generation sequencing, including whole genome sequencing and multimodal retinal imaging including fundus photography, optical coherence tomography (OCT), autofluorescence imaging and adaptive optics (AO) scanning light ophthalmoscopy were performed. Visual electrophysiology was performed in a subset.ResultsEight family members were confirmed as affected by genotyping heterozygous for RDH12 c.763delG. Visual acuity ranged from −0.1 to 0.2 logMAR. Affected individuals had constricted visual fields. A parafoveal and peripapillary ring of hyper-autofluorescence was seen initially, and with progression the area of perifoveal hypo-autofluorescence increased to involve the parafoveal area. Mild retinal thinning was identified on OCT imaging with reduction in both foveal total retinal and outer nuclear layer thickness. Cone densities along the temporal meridian were reduced in affected individuals compared with normative values at all temporal eccentricities studied. One individual with incomplete penetrance, was identified as clinically affected primarily on the basis of AO imaging. Full-field electroretinography demonstrated a rod-cone pattern of dysfunction and large-field pattern electroretinography identified peripheral macular dysfunction.ConclusionsThis novel heterozygous variant RDH12 c.763delG is associated with a rod-cone dystrophy with variable expression. Determination of the degree of penetrance may depend on the modality employed to phenotypically characterise an individual. This rare and specific heterozygous (dominant) variant is predicted to result in a gain of function, that causes disease in a gene typically associated with biallelic (recessive) variants.

show abstract

Molecular cloning and expression of RPE65, a novel retinal pigment epithelium-specific microsomal protein that is post-transcriptionally regulated in vitro

Cited by 287 publications

References 45 publications

Inherited Retinal Diseases Due to RPE65 Variants: From Genetic Diagnostic Management to Therapy

Inherited Retinal Diseases Due to RPE65 Variants: From Genetic Diagnostic Management to Therapy

RPE65 of Retinal Pigment Epithelium, A Putative Receptor Molecule for Plasma Retinol-Binding Protein, is Expressed in Human Keratinocytes

Novel disease-causing variant in RDH12 presenting with autosomal dominant retinitis pigmentosa

Contact Info

Product

Resources

About