April 2010
Volume 51, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2010
Replacement Gene Therapy With a Human RPGRIP1 Sequence Slows Photoreceptor Degeneration in a Murine Model of Leber Congenital Amaurosis
Author Affiliations & Notes
  • B. S. Pawlyk
    Berman-Gund Laboratory, Mass Eye & Ear Infirmary / Harvard Medical School, Boston, Massachusetts
  • O. V. Bulgakov
    Berman-Gund Laboratory, Mass Eye & Ear Infirmary / Harvard Medical School, Boston, Massachusetts
  • X. Liu
    Berman-Gund Laboratory, Mass Eye & Ear Infirmary / Harvard Medical School, Boston, Massachusetts
  • X. Xu
    Berman-Gund Laboratory, Mass Eye & Ear Infirmary / Harvard Medical School, Boston, Massachusetts
  • M. Adamian
    Berman-Gund Laboratory, Mass Eye & Ear Infirmary / Harvard Medical School, Boston, Massachusetts
  • X. Sun
    Berman-Gund Laboratory, Mass Eye & Ear Infirmary / Harvard Medical School, Boston, Massachusetts
  • S. C. Khani
    Schepens Eye Research Institute, Boston, Massachusetts
  • E. L. Berson
    Berman-Gund Laboratory, Mass Eye & Ear Infirmary / Harvard Medical School, Boston, Massachusetts
  • M. A. Sandberg
    Berman-Gund Laboratory, Mass Eye & Ear Infirmary / Harvard Medical School, Boston, Massachusetts
  • T. Li
    Berman-Gund Laboratory, Mass Eye & Ear Infirmary / Harvard Medical School, Boston, Massachusetts
  • Footnotes
    Commercial Relationships  B.S. Pawlyk, None; O.V. Bulgakov, None; X. Liu, None; X. Xu, None; M. Adamian, None; X. Sun, None; S.C. Khani, None; E.L. Berson, None; M.A. Sandberg, None; T. Li, None.
  • Footnotes
    Support  NEI Grant EY010581, NEI core grant P30 EY14104, the Foundation for Retinal Research and the Foundation Fighting Blindness
Investigative Ophthalmology & Visual Science April 2010, Vol.51, 4497. doi:
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      B. S. Pawlyk, O. V. Bulgakov, X. Liu, X. Xu, M. Adamian, X. Sun, S. C. Khani, E. L. Berson, M. A. Sandberg, T. Li; Replacement Gene Therapy With a Human RPGRIP1 Sequence Slows Photoreceptor Degeneration in a Murine Model of Leber Congenital Amaurosis. Invest. Ophthalmol. Vis. Sci. 2010;51(13):4497.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : RPGR-interacting protein 1 (RPGRIP1) is localized in the photoreceptor connecting cilium where it anchors the RPGR (retinitis pigmentosa GTPase regulator) protein. A genetic defect in RPGRIP1 is a known cause of Leber Congenital Amaurosis (LCA), a severe, early-onset form of retinal degeneration that leads to blindness in children. We evaluated the efficacy of replacement gene therapy using a human RPGRIP1 sequence in a murine model of LCA carrying a targeted disruption of RPGRIP1.

Methods: : Cohorts of RPGRIP1 knockout (RPGRIP1-/-) mice up to 5 months of age were studied. At postnatal day 14, mice were given a subretinal injection in one eye of AAV8 vector carrying human RPGRIP1 cDNA under the transcriptional control of a human rhodopsin kinase promoter. Fellow eyes received a GFP vector as control.

Results: : As early as 2 weeks post-injection human RPGRIP1 was expressed specifically in photoreceptors and was localized correctly in the connecting cilia; it also restored the normal localization of RPGR. By immunoblotting analysis human RPGRIP1 (apparent molecular weight - 170 kDa) was found in the treated but not control retinas and matched precisely the endogenous RPGRIP1 protein from human donor retinas. Electroretinogram and histologic examinations showed improved rod and cone photoreceptor function and survival in the treated eyes. Overall, treatment led to a 2-fold improvement in rod function and a 4-fold improvement in cone function as measured by b-wave amplitudes at the final time point. At this stage of disease, control eyes had between 0-2 rows of remaining photoreceptor cells whereas treated eyes typically had between 4-5 rows.

Conclusions: : These data demonstrate efficacy of gene replacement using a human RPGRIP1 cDNA driven by a photoreceptor-specific promoter and establish a prototype vector design for a potential gene therapy in future clinical trials in patients with LCA due to mutations in the gene encoding RPGRIP1.

Keywords: gene transfer/gene therapy • retinal degenerations: hereditary • photoreceptors 
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