The role of mitochondrial DNA mutations and inflammation in adult blindness caused by diabetic retinopathy

糖尿病视网膜病变（DR）是工作年龄人群失明的主要原因，目前尚无治疗方法。线粒体是细胞的细胞器和能量生产的网站在细胞中。线粒体功能障碍参与了DR的发展，我们最近发表了第一项人类研究，表明可以在DR患者的血液样本中检测到MtDNA的变化，并使用患者血液样本和动物/细胞模型发现，糖尿病可导致循环MtDNA升高。由于MtDNA类似于细菌DNA，它可以通过激活TLR 9途径引起炎症反应并激活炎性小体。最近，我们在DR患者中发现了新的MtDNA突变和缺失。在目前的项目中，学生将（a）测试受损的线粒体DNA是视网膜能量不足和循环中慢性炎症的原因的假设（B）使用体外模型验证与DR相关的线粒体变化的时间过程和（c）识别和测试治疗和预防策略。对于第一年的轮换，学生将使用人类患者样本来定义DR中MtDNA突变的发生率。这些研究将进一步扩展，ARPE 19细胞将用于模拟高血糖条件下视网膜中观察到的损伤，以描述特征线粒体变化，然后测试一系列治疗策略。学生将获得一系列细胞生物学和分子生物学技术的培训，包括组织培养，DNA/RNA工作，真实的时间定量PCR，功能测定和生物能量学。

Diabetic retinopathy (DR) is a major cause of blindness in the working-age population for which there is no treatment. Mitochondria are cellular organelles and the site of energy production in the cell. Mitochondrial dysfunction is involved in the development of DR, we recently published the first human study showing that MtDNA changes can be detected in blood samples from DR patients and have found, using patient blood samples and animal/cell models, that diabetes can lead to elevated circulating MtDNA. As MtDNA resembles bacterial DNA it may elicit an inflammatory response via activation of the TLR9 pathway and activate inflammasomes. Recently we have discovered novel MtDNA mutations and deletions in DR patients. In the current project the student will (a) test the hypothesis that damaged MtDNA is causative of energy deficit in the retina and chronic inflammation in circulation (b) characterise the time course of mitochondrial changes associated with DR using in-vitro models and (c) Identify and test therapeutic and preventative strategies. For the rotation/year 1, the student will use human patient samples to define the incidence of MtDNA mutations in DR. These studies will be expanded further and ARPE 19 cells will be used to mimic the damage seen in the retina in hyperglycemic conditions in order to characterise the mitochondrial changes and then test a range of therapeutic strategies. The student will gain training in a range of cell biology and molecular biology techniques including tissue culture, DNA/RNA work, real time quantitative PCR, functional assays and bioenergetics.