June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Triple drug immunosuppression for xenotransplantation of human photoreceptor precursor cells in the canine retina.
Author Affiliations & Notes
  • Ana Ripolles-Garcia
    Division of Experimental Retinal Therapies, Department of Clinical Sciences and Advanced Medicine, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States
  • Natalia Dolgova
    Division of Experimental Retinal Therapies, Department of Clinical Sciences and Advanced Medicine, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States
  • M. Joseph Phillips
    Waisman Center, University of Wisconsin-Madison, Madison, Wisconsin, United States
    McPherson Eye Research Institute, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Svetalana Savina
    Division of Experimental Retinal Therapies, Department of Clinical Sciences and Advanced Medicine, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States
  • Allison Ludwig
    Waisman Center, University of Wisconsin-Madison, Madison, Wisconsin, United States
    McPherson Eye Research Institute, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Sara Stuedemann
    Waisman Center, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • John H Wolfe
    Children’s Hospital of Philadelphia, Philadelphia, Pennsylvania, United States
  • Oliver Garden
    Division of Experimental Retinal Therapies, Department of Clinical Sciences and Advanced Medicine, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States
  • David M Gamm
    Waisman Center, University of Wisconsin-Madison, Madison, Wisconsin, United States
    McPherson Eye Research Institute, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Gustavo D Aguirre
    Division of Experimental Retinal Therapies, Department of Clinical Sciences and Advanced Medicine, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States
  • William A Beltran
    Division of Experimental Retinal Therapies, Department of Clinical Sciences and Advanced Medicine, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States
  • Footnotes
    Commercial Relationships   Ana Ripolles-Garcia None; Natalia Dolgova None; M. Joseph Phillips Opsis Therapeutics, LLC, Code C (Consultant/Contractor); Svetalana Savina None; Allison Ludwig None; Sara Stuedemann None; John H Wolfe None; Oliver Garden None; David Gamm US Patent No. US9752119B2 (OV patent), Code P (Patent), Opsis Therapeutics, LLC, Code S (non-remunerative); Gustavo Aguirre None; William Beltran None
  • Footnotes
    Support  NIH grants U24EY029890, R01EY06855, F30EY031230, Fighting Blindness Canada including philanthropic support from Donna Green and Goldie Feldman, the Foundation Fighting Blindness, Research to Prevent Blindness, the Van Sloun Fund for Canine Genetic Research, Retina Research Foundation Emmett A. Humble Distinguished Directorship of the McPherson Eye Research Institute, and the Sandra Lemke Trout Chair in Eye Research.
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 53 – A0026. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Ana Ripolles-Garcia, Natalia Dolgova, M. Joseph Phillips, Svetalana Savina, Allison Ludwig, Sara Stuedemann, John H Wolfe, Oliver Garden, David M Gamm, Gustavo D Aguirre, William A Beltran; Triple drug immunosuppression for xenotransplantation of human photoreceptor precursor cells in the canine retina.. Invest. Ophthalmol. Vis. Sci. 2022;63(7):53 – A0026.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : To evaluate the effect of a systemic multidrug immunosuppressive protocol on the survival and integration of hESC-derived photoreceptor precursor cells (PRPCs) transplanted into the subretinal space (SRS) of normal and degenerated canine retinas.

Methods : Stage 2 retinal organoids (day 104-151 of differentiation) were generated from a CRX-tdTomato reporter line (WA09 CRX+/tdTomato, WiCell, Madison, WI) and dissociated to produce cell suspension containing fluorescent PRPCs for subretinal injection in 7 normal (12 eyes) and 3 mutant (rcd1/PDE6B) dogs (6 eyes) using a modified subretinal injector (RetinaJect, SurModics, CA). Seven dogs were placed under an immunosuppressive (IS) regimen (prednisolone, cyclosporine A, mycophenolate mofetil), 2 dogs were not medicated, and 1 dog had IS halted after 20 weeks. Survival of PRPCs was monitored by non-invasive multimodal imaging including color and fluorescence photography, cSLO (NIR and BAF modes), cross-sectional and en face OCT. Following termination, survival and integration of PRPCs and involvement of innate and adaptive cellular immune responses were assessed by IHC.

Results : In dogs under IS, loss of transplanted donor cells was seen within the first week post-injection (PI) but remaining cells survived up to 20 weeks PI. In normal retinas, most donor cells remained in the SRS, while in degenerated retinas clusters of cells were found to have migrated into the host’s ONL and inner retina. IHC analysis showed that most PRPCs expressed cone markers (hCA, M/L opsin) and some formed neurites and pedicle-like structures. In dogs that were not under IS, or that had IS halted, there was a rapid loss of transplanted cells. In these dogs, IHC revealed a mixed inflammatory infiltrate composed mostly of macrophages (CD18+) and resident microglia (Iba1+) and to a lesser extent of helper (CD4+) and cytotoxic (CD8+) T-cells as well as B-cells (CD20+).

Conclusions : Our triple drug IS regimen provided long-term (up to 5 months) survival of xenotransplanted hESC-derived PRPCs into the canine retina. Additionally, our results confirmed that degenerating retinas are more suitable for integration of donor PRPCs.

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.

×