June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Successful late-stage disease treatment of P23H human RHO (hRHO) using ARCUS nuclease gene editing in a pig model of Autosomal Dominant Retinitis Pigmentosa (adRP)
Author Affiliations & Notes
  • Archana Jalligampala
    Ophthalmology and Visual Sciences, University of Louisville School of Medicine, Louisville, Kentucky, United States
  • Jennifer M. Noel
    Ophthalmology and Visual Sciences, University of Louisville School of Medicine, Louisville, Kentucky, United States
  • Olivia Noelle Jacobs
    Ophthalmology and Visual Sciences, University of Louisville School of Medicine, Louisville, Kentucky, United States
  • Alec Lynn Bradley
    Ophthalmology and Visual Sciences, University of Louisville School of Medicine, Louisville, Kentucky, United States
  • Wei Wang
    Ophthalmology and Visual Sciences, University of Louisville School of Medicine, Louisville, Kentucky, United States
  • Maha Haleem Jabbar
    Ophthalmology and Visual Sciences, University of Louisville School of Medicine, Louisville, Kentucky, United States
  • Nazarul Hasan
    Anatomical Sciences and Neurobiology, University of Louisville School of Medicine, Louisville, Kentucky, United States
  • Joseph Prestigiacomo
    Ophthalmology and Visual Sciences, University of Louisville School of Medicine, Louisville, Kentucky, United States
  • Jeff Smith
    Precision Biosciences Inc, Durham, North Carolina, United States
  • Victor Bartsevich
    Precision Biosciences Inc, Durham, North Carolina, United States
  • Kristi Viles
    Precision Biosciences Inc, Durham, North Carolina, United States
  • Derek Jantz
    Precision Biosciences Inc, Durham, North Carolina, United States
  • Maureen Ann McCall
    Ophthalmology and Visual Sciences, University of Louisville School of Medicine, Louisville, Kentucky, United States
    Anatomical Sciences and Neurobiology, University of Louisville School of Medicine, Louisville, Kentucky, United States
  • Footnotes
    Commercial Relationships   Archana Jalligampala Precision Biosciences Inc., Code F (Financial Support), Sparing Vision , Code F (Financial Support), Wave Life Sciences , Code F (Financial Support), Nayan Therapeutics, Code F (Financial Support), Rznomics, Code F (Financial Support); Jennifer Noel Precision Biosciences Inc., Code F (Financial Support); Olivia Jacobs Precision Biosciences Inc., Code F (Financial Support); Alec Bradley None; Wei Wang Precision Biosciences Inc., Code F (Financial Support); Maha Jabbar Precision Biosciences Inc., Code F (Financial Support); Nazarul Hasan Precision Biosciences Inc., Code F (Financial Support); Joseph Prestigiacomo Precision Biosciences Inc., Code F (Financial Support); Jeff Smith Precision Biosciences Inc., Code E (Employment); Victor Bartsevich Precision Biosciences Inc., Code E (Employment); Kristi Viles Precision Biosciences Inc., Code E (Employment); Derek Jantz Precision Biosciences Inc., Code O (Owner), Precision Biosciences Inc., Code P (Patent), Precision Biosciences Inc., Code S (non-remunerative); Maureen McCall Precision Biosciences Inc., Code C (Consultant/Contractor), Precision Biosciences Inc., Code F (Financial Support)
  • Footnotes
    Support  EY026158
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 1107. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Archana Jalligampala, Jennifer M. Noel, Olivia Noelle Jacobs, Alec Lynn Bradley, Wei Wang, Maha Haleem Jabbar, Nazarul Hasan, Joseph Prestigiacomo, Jeff Smith, Victor Bartsevich, Kristi Viles, Derek Jantz, Maureen Ann McCall; Successful late-stage disease treatment of P23H human RHO (hRHO) using ARCUS nuclease gene editing in a pig model of Autosomal Dominant Retinitis Pigmentosa (adRP). Invest. Ophthalmol. Vis. Sci. 2022;63(7):1107.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : The P23H mutation in the rhodopsin (RHO) gene represents the most common form of adRP in North America. We previously showed successful gene editing using the ARCUS nuclease, RHO 1-2, a meganuclease, in a TgP23HhRHO pig model of adRP when treatment began at postnatal days, P3-7. This time-point corresponds to mid-stage human adRP, where many rod photoreceptors are present. However, from birth, untreated TgP23H pigs have no rod isolated ffERG response. To determine how late in adRP Rho1-2 gene editing continues to be successful, we evaluated its therapeutic efficacy when treatment began at P18 or P30 when fewer rods remain.

Methods : RHO 1-2 mRNA was packaged in a self-complementary AAV5 vector with a GRK1 promoter and was delivered subretinally (40 µl) to both eyes of TgP23H pigs at P18 and P30. Eyes from untreated animals served as control. The efficacy of 2x1010vg (n:eyes=8, P18; n=6 P30) and 6x1010vg (n=2 eyes, P30) on retinal structure and function were assessed at regular post injection intervals through 20 weeks post-injection (wpi). Fundus imaging, clinical exams, and OCTs assessed RHO 1-2 tolerability and retinal structure. Full-field ERGs assessed retinal function. At final time-point, the animals were euthanized and the retinal sections were processed for immunohistochemistry/confocal microscopy to assess the photoreceptor morphology.

Results : Treatment with RHO 1-2 was well tolerated, with no signs of an immune response. Treatment with both concentrations of RHO 1-2 (2 or 6x1010vg) rejuvenated a rod isolated ffERG b-wave response as early as 8wpi (~9uV). Mean b-wave amplitudes increased until 17wpi (~13-20uV) and were maintained through 20wpi. There is a strong correlation between rod isolated function and rod morphology. RHO 1-2 treated Tg P23H rods, have elongated inner and outer segments (IS/OS) and correctly localized rhodopsin. In contrast, untreated areas in the same retina or untreated Tg retinas have almost no rods and those that remain have no IS/OS and mislocalized rhodopsin.

Conclusions : Our results show that ARCUS nuclease RHO1-2 gene editing of the P23H hRHO allele rejuvenated rods even in late-stage disease. This approach arrested rod degeneration and rejuvenated rod structure and function in TgP23HhRHO pig model of adRP. These findings expand the therapeutic window for RHO 1-2 to retina late-stage adRP.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

×
×

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.

×