iPSC-RPE patch restores photoreceptors and regenerates choriocapillaris in a pig retinal degeneration model
Rohan Gupta, Irina Bunea, Bruno Alvisio, Francesca Barone, Rishabh Gupta, Dara Baker, Haohua Qian, Elena Daniele, Casey G. Contreary, Jair Montford, Ruchi Sharma, Arvydas Maminishkis, Mandeep S. Singh, Maria Teresa Magone De Quadros Costa, Amir H. Kashani, Juan Amaral, Kapil Bharti
Rohan Gupta, Irina Bunea, Bruno Alvisio, Francesca Barone, Rishabh Gupta, Dara Baker, Haohua Qian, Elena Daniele, Casey G. Contreary, Jair Montford, Ruchi Sharma, Arvydas Maminishkis, Mandeep S. Singh, Maria Teresa Magone De Quadros Costa, Amir H. Kashani, Juan Amaral, Kapil Bharti
View: Text | PDF
Research Article Ophthalmology Transplantation

iPSC-RPE patch restores photoreceptors and regenerates choriocapillaris in a pig retinal degeneration model

Abstract

Dry age-related macular degeneration (AMD) is a leading cause of untreatable vision loss. In advanced cases, retinal pigment epithelium (RPE) cell loss occurs alongside photoreceptor and choriocapillaris degeneration. We hypothesized that an RPE-patch would mitigate photoreceptor and choriocapillaris degeneration to restore vision. An induced pluripotent stem cell–derived RPE (iRPE) patch was developed using a clinically compatible manufacturing process by maturing iRPE cells on a biodegradable poly(lactic-co-glycolic acid) (PLGA) scaffold. To compare outcomes, we developed a surgical procedure for immediate sequential delivery of PLGA-iRPE and/or PLGA-only patches in the subretinal space of a pig model of laser-induced outer retinal degeneration. Deep learning algorithm-based optical coherence tomography (OCT) image segmentation verified preservation of the photoreceptors over the areas of PLGA-iRPE–transplanted retina and not in laser-injured or PLGA-only–transplanted retina. Adaptive optics imaging of individual cone photoreceptors further supported this finding. OCT-angiography revealed choriocapillaris regeneration in PLGA-iRPE– and not in PLGA-only–transplanted retinas. Our data, obtained using clinically relevant techniques, verified that PLGA-iRPE supports photoreceptor survival and regenerates choriocapillaris in a laser-injured pig retina. Sequential delivery of two 8 mm2 transplants allows for testing of surgical feasibility and safety of the double dose. This work allows one surgery to treat larger and noncontiguous retinal degeneration areas.

Authors

Rohan Gupta, Irina Bunea, Bruno Alvisio, Francesca Barone, Rishabh Gupta, Dara Baker, Haohua Qian, Elena Daniele, Casey G. Contreary, Jair Montford, Ruchi Sharma, Arvydas Maminishkis, Mandeep S. Singh, Maria Teresa Magone De Quadros Costa, Amir H. Kashani, Juan Amaral, Kapil Bharti

×

Figure 6

OCT-A–based evaluation of choriocapillaris perfusion.

Options: View larger image (or click on image) Download as PowerPoint
OCT-A–based evaluation of choriocapillaris perfusion. (A–F) PLGA-only–tr...
(A–F) PLGA-only–transplanted (A–C) and PLGA-iRPE–transplanted (D–F) laser-injured retinas showing the location of the transplant in fundus infrared images (A and D) and en face OCT-A images of choriocapillaris signal at 50 days (B and E) and 70 days (C and F). (G and H) Fundus infrared image (G) of PLGA-only– and PLGA-iRPE–transplanted retina. The lasered region in A–G is marked with a blue dotted line, PLGA-only with an orange dotted line, and PLGA-iRPE with a solid orange line. The green line in G shows the location of the OCT-A B-scan in H. The corresponding B-scan (H) shows the choriocapillaris signal marked in yellow. (I) Quantification of choriocapillaris signal segmented from OCT-A B-scans under the laser-injured and PLGA-only– and PLGA-iRPE–transplanted retinas. Box and whiskers represent min and max, 25th and 75th percentile, median, and single values. Data were analyzed with a mixed-effect model (REML) and Bonferroni’s multiple comparisons, and 3 images for each eye were analyzed. Number of eyes used for the analysis (I) (n) = 5 T1, 5 T2, and 4 T3 for PLGA-RPE; 4 (T1), 5 (T2), and 4 T3 for PLGA; 7 T1, 7 T2, and 8 T3 for laser. T1 = days 12–28, T2 = days 29–45, T3 = days 46–80. P values are reported as *P < 0.05; **P < 0.01. (J–L) Cross-sectional views of swine retina from laser-treated (J), laser-treated and transplanted with PLGA-iRPE (K), and laser-treated and transplanted with PLGA-only (L), stained for DAPI (blue) to detect nuclei and isolectin (red) to detect choriocapillaris, and immune-stained for Ku80 (pink) to detect human cells. Scale bars: 25 μm.