Differentiation and Transplantation of Embryonic Stem Cell-Derived Cone Photoreceptors into a Mouse Model of End-Stage Retinal Degeneration

• Published in: Stem cell reports 2017-06, Vol.8 (6), p.1659-1674
• Main Authors: Kruczek, Kamil, Gonzalez-Cordero, Anai, Goh, Debbie, Naeem, Arifa, Jonikas, Mindaugas, Blackford, Samuel J.I., Kloc, Magdalena, Duran, Yanai, Georgiadis, Anastasios, Sampson, Robert D., Maswood, Ryea N., Smith, Alexander J., Decembrini, Sarah, Arsenijevic, Yvan, Sowden, Jane C., Pearson, Rachael A., West, Emma L., Ali, Robin R.
• Format: Article
• Language: English
• Subjects: Adaptor Proteins, Signal Transducing - deficiency; Adaptor Proteins, Signal Transducing - genetics; Animals; Basic-Leucine Zipper Transcription Factors - antagonists & inhibitors; Basic-Leucine Zipper Transcription Factors - genetics; Basic-Leucine Zipper Transcription Factors - metabolism; blindness; Cell Differentiation - drug effects; cell- and tissue-based therapy; Disease Models, Animal; Eye Proteins - antagonists & inhibitors; Eye Proteins - genetics; Eye Proteins - metabolism; Hepatocyte Nuclear Factor 6 - metabolism; Leukemia Inhibitory Factor - pharmacology; Mice; Mice, Knockout; mouse embryonic stem cells; Mouse Embryonic Stem Cells - cytology; Mouse Embryonic Stem Cells - transplantation; Oligodendrocyte Transcription Factor 2 - metabolism; Opsins - metabolism; Orphan Nuclear Receptors - antagonists & inhibitors; Orphan Nuclear Receptors - genetics; Orphan Nuclear Receptors - metabolism; Otx Transcription Factors - metabolism; Receptors, Notch - antagonists & inhibitors; Receptors, Notch - metabolism; retina; retinal cone photoreceptor cells; Retinal Cone Photoreceptor Cells - cytology; Retinal Cone Photoreceptor Cells - metabolism; retinal degeneration; Retinal Degeneration - pathology; Retinal Degeneration - therapy; RNA Interference; RNA, Small Interfering - metabolism; Signal Transduction; Tretinoin - metabolism; Tretinoin - pharmacology
• ISSN: 2213-6711; 2213-6711
• DOI: 10.1016/j.stemcr.2017.04.030

The loss of cone photoreceptors that mediate daylight vision represents a leading cause of blindness, for which cell replacement by transplantation offers a promising treatment strategy. Here, we characterize cone differentiation in retinas derived from mouse embryonic stem cells (mESCs). Similar to in vivo development, a temporal pattern of progenitor marker expression is followed by the differentiation of early thyroid hormone receptor β2-positive precursors and, subsequently, photoreceptors exhibiting cone-specific phototransduction-related proteins. We establish that stage-specific inhibition of the Notch pathway increases cone cell differentiation, while retinoic acid signaling regulates cone maturation, comparable with their actions in vivo. MESC-derived cones can be isolated in large numbers and transplanted into adult mouse eyes, showing capacity to survive and mature in the subretinal space of Aipl1−/− mice, a model of end-stage retinal degeneration. Together, this work identifies a robust, renewable cell source for cone replacement by purified cell suspension transplantation. [Display omitted] •Cone photoreceptor precursors form efficiently in mESC retinal organoids•Notch signaling limits temporal competence for cone cell differentiation•Reduced retinoic acid concentrations are required for cone maturation•Cones transplanted into a model of advanced retinal degeneration survive and mature Ali and colleagues characterize the differentiation of cone photoreceptors in mESC retinal organoids, showing substantial numbers of committed cone precursors. Their differentiation is negatively regulated by the Notch pathway, expression of Nrl and Nr2e3, and high concentrations of retinoic acid. Isolated and transplanted into a severely degenerated retina, these cone precursors survive and express markers of advanced maturation.