Mahmoudian-Sani Mohammad-Reza, Forouzanfar Fatemeh, Asgharzade Samira, Ghorbani Nilufar
Thalassemia and Hemoglobinopathy Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
Neuroscience Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
J Ophthalmol. 2019 Dec 11;2019:2454362. doi: 10.1155/2019/2454362. eCollection 2019.
Retinal degeneration is considered as a condition ensued by different blinding disorders such as retinitis pigmentosa, age-related macular degeneration, and diabetic retinopathy, which can cause loss of photoreceptor cells and also lead to significant vision deficiencies. Although there is no efficient treatment in this domain, transplantation of stem cells has been regarded as a therapeutic approach for retinal degeneration. Thus, the purpose of this study was to analyze the potential of human bone marrow-derived mesenchymal stem cells (hBMSCs) to differentiate into photoreceptor cells via transfection of microRNA (miRNA) in vitro for regenerative medicine purposes. To this end, miR-183/96/182 cluster was transfected into hBMSCs; then, qRT-PCR was performed to measure the expression levels of miR-183/96/182 cluster and some retina-specific neuronal genes such as OTX2, NRL, PKC, and recoverin. CRX and rhodopsin (RHO) levels were also measured through qRT-PCR and immunocytochemistry, and subsequently, cellular change morphology was detected. The findings showed no changes in the morphology of the given cells, and the expression of the neuroretinal genes such as OTX2, NRL, and PKC. Moreover, recoverin was upregulated upon miR-183/-96/-182 overexpression in cultured hBMSCs. Ectopic overexpression of the miR-183 cluster could further increase the expression of CRX and RHO at the messenger RNA (mRNA) and protein levels. Furthermore, the data indicated that the miR-183 cluster could serve as a crucial function in photoreceptor cell differentiation. In fact, miRNAs could be assumed as potential targets to exploit silent neuronal differentiation. Ultimately, it was suggested that in vitro overexpression of miR-183 cluster could trigger reprogramming of the hBMSCs to retinal neuron fate, especially photoreceptor cells.
视网膜变性被认为是由不同的致盲疾病引起的一种病症，如色素性视网膜炎、年龄相关性黄斑变性和糖尿病性视网膜病变，这些疾病可导致光感受器细胞丧失，并进而导致严重的视力缺陷。尽管在这一领域尚无有效的治疗方法，但干细胞移植已被视为治疗视网膜变性的一种方法。因此，本研究的目的是分析人骨髓间充质干细胞（hBMSCs）通过体外转染微小RNA（miRNA）分化为光感受器细胞的潜力，以用于再生医学目的。为此，将miR-183/96/182簇转染到hBMSCs中；然后，进行qRT-PCR以测量miR-183/96/182簇以及一些视网膜特异性神经元基因如OTX2、NRL、PKC和恢复蛋白的表达水平。还通过qRT-PCR和免疫细胞化学测量了CRX和视紫红质（RHO）水平，随后检测细胞形态变化。研究结果显示给定细胞的形态以及神经视网膜基因如OTX2、NRL和PKC的表达没有变化。此外，在培养的hBMSCs中，miR-183/-96/-182过表达后恢复蛋白上调。miR-183簇的异位过表达可进一步在信使核糖核酸（mRNA）和蛋白质水平上增加CRX和RHO的表达。此外，数据表明miR-183簇在光感受器细胞分化中可发挥关键作用。事实上，miRNAs可被视为利用沉默神经元分化的潜在靶点。最终，有人提出在体外过表达miR-183簇可触发hBMSCs重编程为视网膜神经元命运，尤其是光感受器细胞。
