Genome engineering of stem cells for kidney regeneration

我们建议开发一种再生基因和细胞疗法来治疗急性肾损伤和/或慢性肾损伤。 肾脏疾病。这些研究是基于从尿液中分离成人干细胞的最新进展。 尿液来源的干细胞有几个显著的特性：它们很容易获得，可以繁殖很多次 通道，是多功能的，也是受伤部位的家园。我们建议对尿液来源的干细胞进行基因组修饰 并在三种小鼠肾损伤模型(缺血再灌注、横纹肌溶解和 马兜铃酸)。细胞将被转座子修饰以表达荧光素酶。这将允许在现场进行跟踪 通过光学成像与先进的层析成像新技术一起研究动物。肾损伤的生物标志物 将被测量和比较。组织学和标记分析将确定干细胞是否对 每种情况下的小鼠表型。目标1旨在比较接受尿源性干细胞治疗的小鼠的结果 细胞与不处理的对比。在目标2中，尿液来源的干细胞将被重新编程为诱导的肾单位祖细胞 细胞，并与过去的诱导肾单位祖细胞的方法进行比较。这些细胞是独一无二的 因为它们与帽间充质中发现的细胞相似，并能够分化为 肾单位。这些目标将决定基于尿液的异基因或自体干细胞移植疗法- 来源干细胞可能具有逆转或延缓急性肾损伤小鼠模型肾损害的能力 受伤或慢性肾脏疾病。

We propose to develop a regenerative gene and cell therapy treatment for acute kidney injury and/or chronic kidney disease. These studies are based on recent advances in isolation of adult stem cells from the urine. Urine-derived stem cells have several remarkable properties: they are easy to obtain, propagate for many passages, are multipotent, and home to sites of injury. We propose to genome modify urine-derived stem cells and track their homing in three mouse models of kidney injury (ischemia reperfusion, rhabdomyolysis, and aristolochic acid). Cells will be modified with transposons to express luciferase. This will permit tracking in live animals by optical imaging together with new techniques for advanced tomography. Biomarkers for kidney injury will be measured and compared. Histology and marker analysis will determine if stem cells have an effect on the mouse phenotype for each condition. Aim 1 seeks to compare outcomes for mice receiving urine-derived stem cells vs no treatment. In Aim 2, urine-derived stem cells will be reprogrammed into induced nephron progenitor cells and compared to past methods of deriving induced nephron progenitor cells. These cells are unique because they are similar to cells found in the cap mesenchyme and are capable of differentiating into all parts of the nephron. These aims will determine if allogeneic or autologous stem cell transplant therapies based on urine- derived stem cells may have the ability to reverse or delay kidney damage in mouse models of acute kidney injury or chronic kidney disease.