Brydon Elizabeth M, Bronstein Revital, Buskin Adriana, Lako Majlinda, Pierce Eric A, Fernandez-Godino Rosario
Department of Ophthalmology, Ocular Genomics Institute, Massachusetts Eye and Ear, Harvard Medical School, Boston, MA, USA.
Institute of Genetic Medicine, Newcastle University, Newcastle upon Tyne, UK.
Mol Ther Methods Clin Dev. 2019 Nov 11;15:392-402. doi: 10.1016/j.omtm.2019.10.014. eCollection 2019 Dec 13.
Retinitis pigmentosa (RP) is the most common form of inherited vision loss and is characterized by degeneration of retinal photoreceptor cells and the retinal pigment epithelium (RPE). Mutations in pre-mRNA processing factor 31 () cause dominant RP via haploinsufficiency with incomplete penetrance. There is good evidence that the diverse severity of this disease is a result of differing levels of expression of the wild-type allele among patients. Thus, we hypothesize that -related RP will be amenable to treatment by adeno-associated virus (AAV)-mediated gene augmentation therapy. To test this hypothesis, we used induced pluripotent stem cells (iPSCs) with mutations in and differentiated them into RPE cells. The mutant iPSC-RPE cells recapitulate the cellular phenotype associated with the PRPF31 pathology, including defective cell structure, diminished phagocytic function, defects in ciliogenesis, and compromised barrier function. Treatment of the mutant iPSC-RPE cells with AAV- restored normal phagocytosis and cilia formation, and it partially restored structure and barrier function. These results suggest that AAV-based gene therapy targeting RPE cells holds therapeutic promise for patients with -related RP.
视网膜色素变性（RP）是遗传性视力丧失最常见的形式，其特征是视网膜光感受器细胞和视网膜色素上皮（RPE）发生退化。前体mRNA加工因子31（）的突变通过单倍剂量不足和不完全外显率导致显性RP。有充分证据表明，这种疾病的不同严重程度是患者中野生型等位基因表达水平不同的结果。因此，我们假设与相关的RP将适合通过腺相关病毒（AAV）介导的基因增强疗法进行治疗。为了验证这一假设，我们使用了具有突变的诱导多能干细胞（iPSC），并将它们分化为RPE细胞。突变的iPSC-RPE细胞重现了与PRPF31病理相关的细胞表型，包括细胞结构缺陷、吞噬功能减弱、纤毛发生缺陷和屏障功能受损。用AAV-处理突变的iPSC-RPE细胞可恢复正常的吞噬作用和纤毛形成，并部分恢复结构和屏障功能。这些结果表明，针对RPE细胞的基于AAV的基因疗法对与相关的RP患者具有治疗前景。
