In vitro transdifferentiation of human peripheral blood mononuclear cells to photoreceptor-like cells.
In vitro transdifferentiation of human peripheral blood mononuclear cells to photoreceptor-like cells.
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DOI：
10.1242/bio.016477
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发表时间：
2016-06-15
期刊：
影响因子：
2.4
通讯作者：
Seko Y
中科院分区：
文献类型：
--
作者：
Komuta Y;Ishii T;Kaneda M;Ueda Y;Miyamoto K;Toyoda M;Umezawa A;Seko Y
直接重编程是一种有前途的、简单的、低成本的从体细胞产生靶细胞的方法，而不使用诱导多能干细胞。最近，外周血单核细胞（PBMC）作为重编程的体细胞来源引起了相当大的关注。作为细胞来源，PBMC在组织收集的容易性方面具有优于真皮成纤维细胞的优势。基于我们的研究涉及通过逆转录病毒载体转导光感受器相关转录因子从人虹膜细胞和人皮肤成纤维细胞产生光敏感光细胞，我们通过仙台病毒载体将这些转录因子转导到PBMC中。我们发现，视网膜疾病相关基因在CRX转导的细胞中被有效地检测到，其中大多数对感光功能至关重要。在功能研究中，在一些CRX转导的细胞中检测到光诱导的内向电流。此外，通过改变培养条件，包括额外的RAX1和NEUROD1的转导，我们发现了比在CRX转导的PBMC中观察到的更多种类的视网膜疾病相关基因。这些数据表明，CRX作为一个主控制基因重编程成光感受器样细胞的PBMC和我们诱导的光感受器样细胞可能有助于个性化的药物筛选和遗传性视网膜变性的疾病建模。总结：我们建立了一种从外周血单个核细胞直接重编程产生光感受器样细胞的方法，用于遗传性视网膜变性的疾病建模。
Direct reprogramming is a promising, simple and low-cost approach to generate target cells from somatic cells without using induced pluripotent stem cells. Recently, peripheral blood mononuclear cells (PBMCs) have attracted considerable attention as a somatic cell source for reprogramming. As a cell source, PBMCs have an advantage over dermal fibroblasts with respect to the ease of collecting tissues. Based on our studies involving generation of photosensitive photoreceptor cells from human iris cells and human dermal fibroblasts by transduction of photoreceptor-related transcription factors via retrovirus vectors, we transduced these transcription factors into PBMCs via Sendai virus vectors. We found that retinal disease-related genes were efficiently detected in CRX-transduced cells, most of which are crucial to photoreceptor functions. In functional studies, a light-induced inward current was detected in some CRX-transduced cells. Moreover, by modification of the culture conditions including additional transduction of RAX1 and NEUROD1, we found a greater variety of retinal disease-related genes than that observed in CRX-transduced PBMCs. These data suggest that CRX acts as a master control gene for reprogramming PBMCs into photoreceptor-like cells and that our induced photoreceptor-like cells might contribute to individualized drug screening and disease modeling of inherited retinal degeneration. Summary: We established a method to generate photoreceptor-like cells from peripheral blood mononuclear cells by direct reprogramming to serve in disease modeling of inherited retinal degeneration.