背景：X连锁幼年视网膜裂（XLRS）是一种由RS1基因突变引起的遗传性疾病，导致视网膜分裂和视力障碍.RS1相关视网膜变性的机制尚不完全清楚。此外,XLRS动物模型在XLRS研究中存在局限性。这里,我们使用人类诱导多能干细胞(hiPSC)衍生的视网膜类器官(ROs)来研究XLRS的疾病机制和潜在治疗方法.
方法：从两个RS1突变体（E72K）XLRS患者的外周血单核细胞重编程的hiPSCs分化为ROs。随后，我们探讨RS1突变是否会影响RO发育,并探讨RS1基因增强治疗的有效性.
结果：来自RS1（E72K）突变hiPSCs的RO在光感受器中表现出发育延迟，视网膜裂素（RS1）缺乏症，与对照RO相比，自发活动改变。此外，发育延迟与杆特异性前体标记(NRL)和光感受器特异性标记(RCVRN)表达降低相关.腺相关病毒（AAV）介导的RS1基因增强在光感受器未成熟阶段挽救了具有RS1（E72K）突变的RO中的杆状光感受器发育延迟。
结论：RS1（E72K）突变导致ROs的光感受器发育延迟，RS1基因增强治疗可以部分挽救。
BACKGROUND: X-linked juvenile retinoschisis (XLRS) is an inherited disease caused by RS1 gene mutation, which leads to retinal splitting and visual impairment. The mechanism of RS1-associated retinal degeneration is not fully understood. Besides, animal models of XLRS have limitations in the study of XLRS. Here, we used human induced pluripotent stem cell (hiPSC)-derived retinal organoids (ROs) to investigate the disease mechanisms and potential treatments for XLRS.
METHODS: hiPSCs reprogrammed from peripheral blood mononuclear cells of two RS1 mutant (E72K) XLRS patients were differentiated into ROs. Subsequently, we explored whether RS1 mutation could affect RO development and explore the effectiveness of RS1 gene augmentation therapy.
RESULTS: ROs derived from RS1 (E72K) mutation hiPSCs exhibited a developmental delay in the photoreceptor, retinoschisin (RS1) deficiency, and altered spontaneous activity compared with control ROs. Furthermore, the delays in development were associated with decreased expression of rod-specific precursor markers (NRL) and photoreceptor-specific markers (RCVRN). Adeno-associated virus (AAV)-mediated gene augmentation with RS1 at the photoreceptor immature stage rescued the rod photoreceptor developmental delay in ROs with the RS1 (E72K) mutation.
CONCLUSIONS: The RS1 (E72K) mutation results in the photoreceptor development delay in ROs and can be partially rescued by the RS1 gene augmentation therapy.
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