Generation of a Transplantable Population of Human iPSC-Derived Retinal Ganglion Cells.

Oriane Rabesandratana,Amélie Slembrouck-Brec,Antoine Mialot,Alain Chédotal,Corentin Joffrois,Gael Orieux,Amélie Rodrigues,Céline Nanteau,Jens Duebel,Antoine Chaffiol,Sacha Reichman,Olivier Goureau,José-Alain Sahel,Giuliana Gagliardi

doi:10.3389/fcell.2020.585675

Abstract

Optic neuropathies are a major cause of visual impairment due to retinal ganglion cell (RGC) degeneration. Human induced-pluripotent stem cells (iPSCs) represent a powerful tool for studying both human RGC development and RGC-related pathological mechanisms. Because RGC loss can be massive before the diagnosis of visual impairment, cell replacement is one of the most encouraging strategies. The present work describes the generation of functional RGCs from iPSCs based on innovative 3D/2D stepwise differentiation protocol. We demonstrate that targeting the cell surface marker THY1 is an effective strategy to select transplantable RGCs. By generating a fluorescent GFP reporter iPSC line to follow transplanted cells, we provide evidence that THY1-positive RGCs injected into the vitreous of mice with optic neuropathy can survive up to 1 month, intermingled with the host RGC layer. These data support the usefulness of iPSC-derived RGC exploration as a potential future therapeutic strategy for optic nerve regeneration.

Highlights

Retinal ganglion cells (RGCs) play a major role in the visual function in transmitting visual information from the retina through the optic nerve and optic tract to the brain structures dedicated to processing visual information
Based on our good manufacturing practice (GMP)-compliant retinal differentiation protocol (Reichman et al, 2017), we demonstrate that RGCs cultured in 2D conditions after dissociation of early retinal organoids derived from human induced pluripotent stem cells (hiPSCs) strongly express the cell surface antigen THY1
Because THY1 is considered as a specific marker of RGC in a retinal context (Barnstable and Dräger, 1984), we first sought to determine, using our previously GMP-compatible protocol (Reichman et al, 2017), the expression of this cell surface marker during the differentiation of hiPSC-derived retinal organoids in floating conditions

Summary

Introduction

Retinal ganglion cells (RGCs) play a major role in the visual function in transmitting visual information from the retina through the optic nerve and optic tract to the brain structures dedicated to processing visual information. RGC impairment is a common feature in many pathologies leading to visual loss, generally referred to as optic neuropathies such as Leber’s hereditary optic neuropathy (LHON), and dominant optic atrophy or glaucoma, which is the second cause of blindness in the world (Jonas et al, 2017). Despite treatments, RGC degeneration and visual loss still progress in some patients. Retinal Ganglion Cell Differentiation From iPSCs leading to irreversible blindness (Susanna et al, 2015; Greco et al, 2016). RGC loss mainly occurs in young patients, notably LHON-patients with no effective treatment to date (Newman, 2012; Carelli et al, 2017)

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