Retinal Muller Glial Cells in the initiation of diabetic retinopathy

项目摘要/摘要 糖尿病视网膜病变(DR)发病的分子机制及原发细胞 糖尿病在视网膜中的作用靶点尚未完全阐明。这是一个重大障碍， 开发有效的治疗方法来预防或减缓疾病的发生。当受到以下挑战时 糖尿病、视网膜神经元、神经胶质细胞和血管系统都表现出异常。即使它目前不是 明确哪些细胞类型是糖尿病的主要靶点，Müller胶质细胞(MG)作为第一反应细胞之一 糖尿病在视网膜中的作用，对糖尿病视网膜病变的发展是必不可少的。然而，分子 控制糖尿病引起的Müler神经胶质反应的机制仍未得到充分研究。 我们应用单细胞转录分析(单细胞RNA-SEQ)系统地描述了糖尿病- 诱导糖尿病大鼠模型视网膜多细胞反应(初步研究)。在53种类型中 单细胞RNA-seq检测到视网膜细胞，MG是糖尿病最早的反应者之一 转录水平。值得注意的是，MG最初上调了在其他系统中发挥保护作用的基因， 包括抗凋亡、抗增殖、抗氧化、抗炎等基因，但未能维持 随着疾病的发展，这些保护性基因的表达水平。这一失败可能会导致 我们假设MG通过上调保护性基因发挥保护作用 早期的DR，加强这种内在的保护机制将保护视网膜免受糖尿病的影响- 诱导性损害。 拟议的研究将在两个目标上检验这一假设。在目标1中，我们将重点研究以下其中之一 候选保护基因，锌指蛋白36同源基因(Zfp36)，最初被糖尿病上调 在MG中，然后在糖尿病大鼠模型中，随着DR的进展而下调。在目标2中，我们将确定 使用基于CRISPR的新技术在MG中多路激活保护通路是否可以 进一步保护视网膜免受糖尿病引起的损伤。 总之，这项拟议的研究旨在揭示MG在启动DR中的作用，重点是解剖 它们的保护作用。这项工作将有助于更好地了解DR和新的治疗候选药物。

PROJECT SUMMARY/ABSTRACT The molecular mechanisms responsible for the initiation of diabetic retinopathy (DR), and the primary cellular targets of diabetes in the retina have not been fully elucidated. This represents a significant barrier to the development of effective therapies to prevent or slow down the initiation of the disease. When challenged by diabetes, retinal neurons, glia, and the vasculature all display abnormalities. Even though it is currently not clear which cell types are the primary targets of diabetes, Müller glial cells (MG), as one of the first responders of diabetes in the retina, are essential for the development of diabetic retinopathy. However, the molecular mechanisms controlling the diabetes-induced Müller glial responses remain understudied. We applied single cell transcriptomic analysis (single cell RNA-seq) to systematically profile diabetes- induced multicellular responses in the retina of diabetic rat models (preliminary studies). Among the 53 types of retinal cell detected by single cell RNA-seq, MG were one of the first responders to diabetes at the transcriptional level. Notably, MG initially upregulated genes that play protective roles in other systems, including anti-apoptosis, anti-proliferation, anti-oxidation, and anti-inflammation genes, but failed to maintain expression levels of these protective genes as the disease progressed. This failure could contribute to the development of DR. We hypothesize that MG exert protective roles by upregulating protective genes in the early stage of DR, and that enhancing this intrinsic protective mechanism will protect the retina from diabetes- induced damage. The proposed studies will test this hypothesis in two aims. In Aim 1, we will focus on studying one of the candidate protective genes, Zinc finger protein 36 homolog (Zfp36), which was initially upregulated by diabetes in MG and then downregulated as DR progressed, using diabetic rat models. In Aim 2, we will determine whether multiplexing activation of protective pathways in MG with a novel CRISPR-based technique can further protect the retina from diabetes-induced damage. In summary, the proposed study aims to uncover the roles of MG in initiating DR, focusing on dissecting their protective effects. This work will lead to better understanding of DR and new therapeutic candidates.