人胚胎干细胞来源视网膜色素上皮细胞通过CX3CL1通路调控小胶质细胞活化的作用机制研究

Transplantation of human embryonic stem cells derived- retinal pigment epithelium cell (hESC-RPE) in the treatment of retinal degeneration, including AMD, has been proceed to clinical trials. Regulation the transplanted hESC-RPE cell to achieve best biology function is important for the clinical application of stem cell-based treatment. Microglia is the resident macrophage cell in the retina, which plays neuroprotective effect and maintain retinal homeostasis. Activation of microglia induce retinal damage through release of inflammation cytokines. Our previous works have demonstrated that microglias were activated by retinal degeneration and suppressed during sub-retinal implantation of hESC-RPE, of which, the specific mechanism is not quite clear. In this study, we will build pGC-FU-hCX3CL1 and pGC-FU-hCX3CL1 siRNA lentiviral vector plasmid and transfect hESC-RPE cells. Furthermore, the suppression of MG activation by manipulating the CX3CL1-CX3CR1 signaling pathway will be investigated, as well as biological function of stem cells in cells and in animal models. This study will help to understand the molecular mechanism of interaction of hESC-RPE and MG, and further help to provide potential treatment strategy of stem cell transplantation combined with gene therapy in retinal degenerative diseases.

人胚胎干细胞来源视网膜色素上皮细胞（hESC-RPE）移植治疗AMD等视网膜变性疾病已进入临床试验，研究如何调控移植的hESC-RPE细胞发挥生物学功能及具体机制对于干细胞治疗的临床应用具有重要价值。小胶质细胞（MG）是在视网膜定居、有神经保护功能的吞噬细胞，参与维护视网膜微环境稳定；活化MG通过释放炎症因子加剧视网膜损伤。我们的前期研究证实视网膜变性存在MG活化，视网膜下hESC-RPE移植能够抑制MG活化，但具体分子机制并不明确。本课题拟构建pGC-FU-hCX3CL1和pGC-FU-hCX3CL1 siRNA慢病毒载体质粒，并转染hESC-RPE细胞；通过CX3CL1-CX3CR1信号通路调控MG活化，并在细胞及动物模型上研究对干细胞生物学功能及视网膜微环境的作用。本研究有助于阐明hESC-RPE和MG之间的相互作用及机制，为视网膜变性疾病的干细胞移植联合基因治疗提供靶点。

各种干细胞来源的视网膜色素上皮细胞移植治疗年龄相关性黄斑变性等视网膜变性疾病已进入临床试验，研究如何调控移植视网膜色素上皮细胞发挥生物学功能及具体机制对于疾病治疗的临床应用具有重要价值。小胶质细胞是在视网膜定居、有神经保护功能的吞噬细胞，参与维护视网膜微环境稳定；活化小胶质细胞通过释放炎症因子加剧视网膜损伤。我们的前期研究证实视网膜变性存在小胶质细胞活化，视网膜下移植的视网膜色素上皮细胞能够抑制小胶质细胞活化，但具体分子机制并不明确。本课题拟通过构建CX3CL1 siRNA慢病毒载体质粒，并转染视网膜色素上皮细胞，证实视网膜色素上皮细胞通过CX3CL1-CX3CR1信号通路调控小胶质细胞活化，本研究有助于阐明视网膜色素上皮细胞和小胶质细胞之间的相互作用及机制，为视网膜变性疾病的干细胞移植联合基因治疗提供靶点。