Journal Article

Acquisition of neurodegenerative features in isogenic OPTN(E50K) human stem cell-derived retinal ganglion cells associated with autophagy disruption and mTORC1 signaling reduction

Shiga, Yukihiro, Harkin, Jade, VanderWall, Kirstin B., Di Polo, Adriana, Gomes, Cátia, Fligor, Clarisse M., Hetzer, Shelby M., Lavekar, Sailee S., Meyer, Jason S., Huang, Kang-Chieh, Belforte, Nicolas, Patil, Shruti V.

2024

Overview

The ability to derive retinal ganglion cells (RGCs) from human pluripotent stem cells (hPSCs) has led to numerous advances in the field of retinal research, with great potential for the use of hPSC-derived RGCs for studies of human retinal development, in vitro disease modeling, drug discovery, as well as their potential use for cell replacement therapeutics. Of all these possibilities, the use of hPSC-derived RGCs as a human-relevant platform for in vitro disease modeling has received the greatest attention, due to the translational relevance as well as the immediacy with which results may be obtained compared to more complex applications like cell replacement. While several studies to date have focused upon the use of hPSC-derived RGCs with genetic variants associated with glaucoma or other optic neuropathies, many of these have largely described cellular phenotypes with only limited advancement into exploring dysfunctional cellular pathways as a consequence of the disease-associated gene variants. Thus, to further advance this field of research, in the current study we leveraged an isogenic hPSC model with a glaucoma-associated mutation in the Optineurin (OPTN) protein, which plays a prominent role in autophagy. We identified an impairment of autophagic-lysosomal degradation and decreased mTORC1 signaling via activation of the stress sensor AMPK, along with subsequent neurodegeneration in OPTN(E50K) RGCs differentiated from hPSCs, and have further validated some of these findings in a mouse model of ocular hypertension. Pharmacological inhibition of mTORC1 in hPSC-derived RGCs recapitulated disease-related neurodegenerative phenotypes in otherwise healthy RGCs, while the mTOR-independent induction of autophagy reduced protein accumulation and restored neurite outgrowth in diseased OPTN(E50K) RGCs. Taken together, these results highlighted that autophagy disruption resulted in increased autophagic demand which was associated with downregulated signaling through mTORC1, contributing to the degeneration of RGCs.

• Publisher: BioMed Central,BioMed Central Ltd,BMC
• Subject: Animals / Autophagy (Cytology) / Autophagy - physiology / Biomedical and Life Sciences / Biomedicine / Cell Cycle Proteins - genetics / Cell Cycle Proteins - metabolism / Degeneration / Health aspects / Humans / Mechanistic Target of Rapamycin Complex 1 - genetics / Mechanistic Target of Rapamycin Complex 1 - metabolism / Membrane Transport Proteins - genetics / Membrane Transport Proteins - metabolism / Mice / Nervous system / Neurology / Neuropathology and Neurodegeneration in the Retina / Neurosciences / Pathology / Physiological aspects / Pluripotent Stem Cells / Protein kinases / Retina / Retinal ganglion cells / Retinal Ganglion Cells - metabolism / Retinal Ganglion Cells - pathology / Signal Transduction - physiology / Stem cells