Human iPSC-based retinal cell library for disease modeling and drug discovery

Abstract

Purpose : The possibility to make a “Disease-in-a-Dish” using patient-derived induced pluripotent stem cells (iPSCs) represent an unexpected opportunity to elaborate useful research tools for drug development. Part of the challenge in cell-based drug discovery is the production of large number of identical cells of interest. In this way, we present here innovative cell-based approaches for large production and banking of specific epithelial, glial or neuronal retinal cells to model retinal dystrophies as retinitis pigmentosa (RP), diabetic retinopathy (DR) or age-related macular degeneration (AMD).

Methods : Defined culture of adherent human iPSC (hiPSCs), floating hiPSC-derived retinal organoids (ROs) and RO-isolated retinal progenitor cells (hiRPCs) were used to produce all retinal cells types. HiPSC-derived retinal pigmented epithelial cells (hiRPE) were produced directly from adherent hiPSC cultures. HiPSCs-derived Muller glial cells (hiMGCs) were selected and expanded from mature ROs. Expandable and multipotent hiRPCs were selected from early ROs using innovative culture conditions and RPC-dedicated medium.

Results : Both hiRPE, hiMGCs and multipotent hiRPCs can be cryopreserved while retaining their proliferation capacity for large-scale productions. The development of a hiRPE-based model of retinitis pigmentosa coupled to high throughput screening (HTS) identified potential repositionable drugs from the FDA-approved compound library. HiMGCs recapitulated inflammatory and angiogenic features of DR in pathological-like culture condition compatible with HTS. Different neuroretinal cells such as photoceptor precursors, ganglion, amacrine and horizontal cells can be easily differentiated from previously expanded hiRPCs. Moreover, specific culture conditions to direct photoreceptor differentiation of hiRPCs allowed the production of large and high enriched cultures of photoceptor precursors (>90%) in 2 weeks without purification step.

Conclusions : Large-scale productions of relevant human retinal cells such as hiRPEs, hiMGCs, hiRPCs and photoreceptor precursors are now available. This retinal cell library offers new biological tools to better understand the different retinal dystrophies and to develop new treatments adapted to each type of the disease.

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