Concise Review: Making Stem Cells Retinal: Methods for Deriving Retinal Pigment Epithelium and Implications for Patients With Ocular Disease

Leach, Lyndsay L.; Clegg, Dennis O.

doi:10.1002/stem.2010

Cited by 49 publications

(35 citation statements)

References 86 publications

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“…Transplantation of stem cell-derived RPE has emerged as a possibility for treating AMD (12,13), and clinical trials are currently underway (e.g. (14)(15)(16)(17)(18)).…”

Section: /Body Introductionmentioning

confidence: 99%

Regeneration of the zebrafish retinal pigment epithelium after widespread genetic ablation

Hanovice

Leach

Slater

et al. 2018

Preprint

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The retinal pigment epithelium (RPE) is a specialized monolayer of pigmented cells within the eye that is critical for maintaining visual system function. Diseases affecting the RPE have dire consequences for vision, and the most prevalent of these is atrophic (dry) age-related macular degeneration (AMD), which is thought to result from RPE dysfunction and degeneration. An intriguing possibility for treating RPE degenerative diseases like atrophic AMD is the stimulation of endogenous RPE regeneration; however, very little is known about the mechanisms driving successful RPE regeneration in vivo. Here, we developed a zebrafish transgenic model (rpe65a:nfsB-GFP) that enabled ablation of large swathes of mature RPE. RPE ablation resulted in rapid RPE degeneration, as well as degeneration of Bruch's membrane and underlying photoreceptors. Using this model, we demonstrate for the first time that larval and adult zebrafish are capable of regenerating a functional RPE monolayer after RPE ablation. Regenerated RPE cells first appear at the periphery of the RPE, and regeneration proceeds in a peripheral-to-central fashion. RPE ablation elicits a robust proliferative response in the remaining RPE. Subsequently, proliferative cells move into the injury site and differentiate into RPE. BrdU pulse-chase analyses demonstrate that the regenerated RPE is likely derived from remaining peripheral RPE cells. Pharmacological inhibition of Wnt signaling significantly reduces cell proliferation in the RPE and delays overall RPE recovery. These data demonstrate that the zebrafish RPE possesses a robust capacity for regeneration and highlight a potential mechanism through which endogenous RPE regenerate in vivo. SIGNIFICANCE STATEMENTDiseases resulting in RPE degeneration are among the leading causes of blindness worldwide, and no therapy exists that can replace RPE or restore lost vision. One intriguing possibility is the development of therapies focused on stimulating endogenous RPE regeneration. For this to be possible, we must first gain a deeper understanding of the mechanisms underlying RPE regeneration. Here, we ablate mature RPE in zebrafish and demonstrate that zebrafish regenerate RPE after widespread injury. Injury-adjacent RPE proliferate and regenerate RPE, suggesting that they are the source of regenerated tissue. Finally, we demonstrate that Wnt signaling is required for RPE regeneration. These findings establish an in vivo model through which the molecular and cellular underpinnings of RPE regeneration can be further characterized.

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“…Transplantation of stem cell-derived RPE has emerged as a possibility for treating AMD (12,13), and clinical trials are currently underway (e.g. (14)(15)(16)(17)(18)).…”

Section: /Body Introductionmentioning

confidence: 99%

Regeneration of the zebrafish retinal pigment epithelium after widespread genetic ablation

Hanovice

Leach

Slater

et al. 2018

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…Although several protocols describing differentiation of RPE from human embryonic stem cells (hESC) and human induced pluripotent stem cells (hiPSC) have been described, they often have limitations, including low yield, high variability, or steps involving embryoid bodies, cysts, or eye cup structures, together with manual excision of pigmented cells , . These limitations are undesirable for scale‐up and transfer to a good manufacturing practice (GMP) clinical manufacturing setting.…”

Section: Introductionmentioning

confidence: 99%

Directing Differentiation of Pluripotent Stem Cells Toward Retinal Pigment Epithelium Lineage

Choudhary

Booth

Gutteridge

et al. 2016

Stem Cells Translational Medicine

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Development of efficient and reproducible conditions for directed differentiation of pluripotent stem cells into specific cell types is important not only to understand early human development but also to enable more practical applications, such as in vitro disease modeling, drug discovery, and cell therapies. The differentiation of stem cells to retinal pigment epithelium (RPE) in particular holds promise as a source of cells for therapeutic replacement in age‐related macular degeneration. Here we show development of an efficient method for deriving homogeneous RPE populations in a period of 45 days using an adherent, monolayer system and defined xeno‐free media and matrices. The method utilizes sequential inhibition and activation of the Activin and bone morphogenetic protein signaling pathways and can be applied to both human embryonic stem cells and induced pluripotent stem cells as the starting population. In addition, we use whole genome transcript analysis to characterize cells at different stages of differentiation that provides further understanding of the developmental dynamics and fate specification of RPE. We show that with the described method, RPE develop through stages consistent with their formation during embryonic development. This characterization— together with the absence of steps involving embryoid bodies, three‐dimensional culture, or manual dissections, which are common features of other protocols—makes this process very attractive for use in research as well as for clinical applications. Stem Cells Translational Medicine 2017;6:490–501

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“…In recent years, several protocols for directional differentiation of ESC and IPSC into RPE have been developed and some of them have been tested in clinical trials [5,[35][36][37][38][39]. It should be noted that the anatomical and morphological features of the eye (relatively small size, organ pairing, well-developed methods of diagnosis and instrumental monitoring, possible immune privilege and presence of the hematoretinal barrier) make it a convenient target for refining the technology of delivery of material in the case of cell therapy involving pluripotent cells derivatives [40].…”

Section: Potential Of Cellular Therapy In Diseases Of the Eyementioning

confidence: 99%

Possibilities for Using Pluripotent Stem Cells for Restoring Damaged Eye Retinal Pigment Epithelium

et al. 2018

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The retinal pigment epithelium is a monolayer of pigmented, hexagonal cells connected by tight junctions. These cells compose part of the outer blood-retina barrier, protect the eye from excessive light, have important secretory functions, and support the function of photoreceptors, ensuring the coordination of a variety of regulatory mechanisms. It is the degeneration of the pigment epithelium that is the root cause of many retinal degenerative diseases. The search for reliable cell sources for the transplantation of retinal pigment epithelium is of extreme urgency. Pluripotent stem cells (embryonic stem or induced pluripotent) can be differentiated with high efficiency into the pigment epithelium of the retina, which opens up possibilities for cellular therapy in macular degeneration and can slow down the development of pathology and, perhaps, restore a patient's vision. Pioneering clinical trials on transplantation of retinal pigment epithelial cells differentiated from pluripotent stem cells in the United States and Japan confirmed the need for developing and optimizing such approaches to cell therapy. For effective use, pigment epithelial cells differentiated from pluripotent stem cells should have a set of functional properties characteristic of such cells in vivo. This review summarizes the current state of preclinical and clinical studies in the field of retinal pigment epithelial transplantation therapy. We also discuss different differentiation protocols based on data in the literature and our own data, and the problems holding back the widespread therapeutic application of retinal pigment epithelium differentiated from pluripotent stem cells.

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Concise Review: Making Stem Cells Retinal: Methods for Deriving Retinal Pigment Epithelium and Implications for Patients With Ocular Disease

Cited by 49 publications

References 86 publications

Regeneration of the zebrafish retinal pigment epithelium after widespread genetic ablation

Regeneration of the zebrafish retinal pigment epithelium after widespread genetic ablation

Directing Differentiation of Pluripotent Stem Cells Toward Retinal Pigment Epithelium Lineage

Possibilities for Using Pluripotent Stem Cells for Restoring Damaged Eye Retinal Pigment Epithelium

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