CRB1 in the human retina in vitro: A functional, pathomechanistical and developmental study using Human Induced Pluripotent Stem Cell-derived retinal organoids
Molecular Biology and Physiology of Neurons and Glial Cells
Molecular and Cellular Neurology and Neuropathology
Final Report Abstract
Retinitis pigmentosa is a devastating genetic eye disease which leads to strong loss of vision or even complete blindness in affected persons. It affects and destroys the retina, the light-sensitive part of the eye. Until now, there is still no treatment option which could cure or even treat affected patients which is why there is a great need in understanding the mechanisms of the disease and to create models of the disease which can be used for developing and testing of therapies. One of the most promising models to study diseases that affect the human retina are so called retinal organoids. Such organoids are tiny three-dimensional tissues that can be derived from every individual. This is made possible by so-called induced pluripotent stem cells which can be easily generated from donor blood or even hair roots. Retinal organoids contain all major features of a retina: The same retinal cell types, tissue organization and functionality. Hence, the are an optimal model to study the changes that occur in patients without the need of animal testing. In this study, we made use of retinal organoids which were obtained from two retinitis pigmentosa patients harboring a mutation in a gene called CRB1. CRB1 is an important protein for the cell organization and maintenance of the light-sensitive cells of the retina, the photoreceptors. Analyzing those organoids, we found that the CRB1 protein is absent in its natural position inside the retina. This was a surprising find since in patients the protein only has a single mutated base. Furthermore, we found that those organoids show changes in major pathway that are important for retinal development. Finally, we the photoreceptors of patient organoids react differently on light stimulation than comparable controls. The obtained results represent a thorough characterization of CRB1 patient organoids and give the baseline for further in-depth characterization and the possible development of a therapy. Indeed, based on the results, we developed a therapy approach is based on a novel RNA technology. In the future, the knowledge about the disease as well as the development of therapy approaches could contribute to the improvement of Retinitis Pigmentosa patient situation and deepen our biological knowledge of the human retina.
Publications
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2017. Microphysiological organoid culture. DE 10 2017 217 738. PCT/EP2018/076645. CA3078426A1
Achberger K, Loskill P, Haderspeck J, Probst C, Liebau S
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Human Retina-on-a-Chip: “Merging Organoid and Organ-on-a-Chip Technology to Generate Complex Multi-Layer Tissue Models.” Elife, vol. 8, Aug. 2019
Achberger K, Probst C, Haderspeck JC, Rogal J, Chuchuy J, Nikolova M, Cora V, Haq W, Bolz S, Shen N, Schenke-Layland K, Ueffing M, Liebau S, Loskill P
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“A Cleared View on Retinal Organoids” Cells, vol. 8, no. 5, p. 391, Apr. 2019
Cora V, Haderspeck J, Antkowiak L, Mattheus U, Neckel PH, Mack AF, Bolz S, Ueffing M, Pashkovskaia N, Achberger K and Liebau
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“Mikrophysiologisches Retinamodell als Alternative zu Tiermodellen“, BIOspektrum 26, 402-405, 2020
Chuchuy J, Achberger K, Liebau S & and Loskill P
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“Generation of Functional Vascular Endothelial Cells and Pericytes from Keratinocyte Derived Human Induced Pluripotent Stem Cells.” Cells. 5;10(1):74. Jan 2021
Pars S, Achberger K, Kleger A, Liebau S, Pashkovskaia N
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“Using transcriptomic analysis to assess double- strand break repair activity: Towards precise in vivo genome editing.” International Journal of Molecular Sciences, 21(4), 2021
Pasquini G, Cora V, Swiersy A, Achberger K, Antkowiak L, Müller B, Wimmer T, Fraschka SAK, Casadei N, Ueffing M, Liebau S, Stieger K and Busskamp V
