June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
In Vivo Gene Augmentation in the ADAMTS10-Mutant Canine Trabecular Meshwork (TM) with Y444F-Capsid Mutant, Single-Stranded AAV2
Author Affiliations & Notes
  • Christine Harman
    Michigan State University, East Lansing, MI
  • Annie Oh
    Michigan State University, East Lansing, MI
    Western University of Health Sciences, Pomona, CA
  • Kristin L Koehl
    Michigan State University, East Lansing, MI
  • Sanford L Boye
    Ophthalmology & Powell Gene Therapy Center, University of Florida, Gainesville, FL
  • Vince Chiodo
    Ophthalmology & Powell Gene Therapy Center, University of Florida, Gainesville, FL
  • Jiayan Huang
    Ophthalmology, School of Medicine, University of Pennsylvania, Philadelphia, PA
  • Gui-Shuang Ying
    Ophthalmology, School of Medicine, University of Pennsylvania, Philadelphia, PA
  • Shannon Elizabeth Boye
    Ophthalmology & Powell Gene Therapy Center, University of Florida, Gainesville, FL
  • William W Hauswirth
    Ophthalmology & Powell Gene Therapy Center, University of Florida, Gainesville, FL
  • Andras M Komaromy
    Michigan State University, East Lansing, MI
    School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA
  • Footnotes
    Commercial Relationships Christine Harman, None; Annie Oh, None; Kristin Koehl, None; Sanford Boye, PCT/US2012/062478 (P); Vince Chiodo, None; Jiayan Huang, None; Gui-Shuang Ying, None; Shannon Boye, None; William Hauswirth, AGTC (C), AGTC (I), Bionic Sight (I); Andras Komaromy, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 3648. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to Subscribers Only
      Sign In or Create an Account ×
    • Get Citation

      Christine Harman, Annie Oh, Kristin L Koehl, Sanford L Boye, Vince Chiodo, Jiayan Huang, Gui-Shuang Ying, Shannon Elizabeth Boye, William W Hauswirth, Andras M Komaromy; In Vivo Gene Augmentation in the ADAMTS10-Mutant Canine Trabecular Meshwork (TM) with Y444F-Capsid Mutant, Single-Stranded AAV2. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):3648.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: We have previously reported robust targeting of GFP reporter gene expression to the rodent and canine TM with capsid-mutant AAV2(Y444F), both self-complementary and single-stranded. The purpose of our study was to use this vector construct for in vivo TM-targeted gene augmentation in order to prevent/reverse intraocular pressure (IOP) elevation in dogs with primary open-angle glaucoma (POAG) due to a G661R missense mutation in ADAMTS10.

Methods: Eleven adult, ADAMTS10-mutant dogs (mean age: 10.9 ± 4.8 months) were treated unilaterally by intracameral injection of single-stranded AAV2(Y444F)-smCBA-hADAMTS10 vector with hADAMTS10 expression under the constitutive small CMV-chicken beta-actin (smCBA) promoter. Ten dogs received 50-μL and 1 dog received 150 µL of vector solution at a concentration of 2 x 1012 vg/mL. The fellow eyes served as non-injected controls. The dogs were closely followed over 6 months by routine ophthalmic examination and weekly diurnal IOP measurements with a rebound tonometer (Icare TONOVET). IOPs were compared between treated and non-treated eyes by use of a generalized linear model and estimating equation. hADAMTS10 mRNA expression in the treated TM was evaluated by qPCR and compared to untreated control eyes.

Results: The AAV2 vector was well tolerated with no detectable adverse effects. Consistent with our previous findings using single stranded AAV2(Y444F) vector expressing GFP reporter gene, hADAMTS10 transgene expression was detected in the TM of AAV-treated but not in untreated eyes. Unfortunately, no therapeutic IOP effect was observed. Characteristic for the ADAMTS10-POAG disease phenotype, the gradual increase in IOP continued in both treated and untreated eyes during the 6-month follow-up. Among 7 of the study dogs at a comparable POAG disease stage no significant difference in average IOP was observed between treated (25.0 mmHg ± 1.0 mmHg) and untreated control eyes (24.4 mmHg ± 1.2 mmHg) over the 6-month post-treatment study period (p=0.56).

Conclusions: The intracameral administration of AAV2(Y444F) at the quantity administered was safe and resulted in TM-transgene expression but did not achieve a rescue effect. Possible explanations for this treatment failure are insufficient transfection efficiency and/or irreversibility of TM fibrosis/plaque formation in the POAG-affected eyes.

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×