(W1311) GENERATION OF IPSC LINES FROM PATIENTS WITH BARDET BIEDL SYNDROME AND STARGARDT DISEASE AS IN VITRO HUMAN MODELS OF INHERITED RETINAL DYSTROPHY

Abstract: Inherited Retinal dystrophies (IRDs) are a group of disorders involving the degeneration of photoreceptor cells or retinal pigment epithelia leading to vision loss or legal blindness. IRDs are clinically and genetically heterogeneous, with 271 associated genes reported. Therefore, the recapitulation of IRDs has been challenging especially in light of complexities associated with the translation of findings from animal models to human context. Retinal organoid (RO) technology derived from induced pluripotent stem cells (iPSCs) has been an indispensable alternative to closely mimic in vivo systems in order to study the disease pathophysiology. However, many IRDs are underexplored due to the diseases´ genetic heterogeneity and the limited availability of iPSC lines. In this study, we reprogrammed peripheral blood mononuclear cells from two patients suffering from Bardet Biedel Syndrome (BBS-POMGNT1, c.1539+1G>A) and Stargardt disease (STGD1, ABCA4, c.5917del.), respectively using Sendai viruses. Two stem cell lines harboring the patients´ mutations were generated from this approach. Alkaline phosphatase treatment and immunostainings using OCT-4, TRA1-60 and SOX2 were utilized to confirm the pluripotency of the generated cell lines. Germ layer differentiation was performed and characterization was done using endodermal, mesodermal, and ectodermal markers. We confirmed the lines’ genome integrity using karyotyping and the presence of the mutations by genetic sequencing. We subsequently generated ROs using our already established RO protocol confirming the ability of the lines to generate retinal tissue. We aim next to generate isogenic control lines and then retino-cortical assembloid models to better recapitulate and understand the disease pathophysiology.

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