Mesenchymal stem cell senescence in diabetic nephropathy (K23 COVID Admin Supplement)

项目总结/摘要 我是马约诊所的执业肾病学家，我将利用我在临床和 流行病学背景，将拟议的基础研究调查纳入真正的转化研究 糖尿病肾病（DN）患者的治疗。我已经组建了一个多学科的导师团队， 提供开展拟议研究所需的指导、基础设施和工具。我的直系 职业目标是获得实验室研究技能，临床试验经验，再生医学基金， 知识和再生医学课程。我的长期目标是成为再生能源的领导者 肾脏科和一名独立的、富有成效的临床医生-研究者。我的职业发展计划将为我提供 有了过渡到独立所需的基础。我得到了部门最有力的支持， 部门和机构。在拟议的研究中，我将探索一种新的治疗平台的可行性 我相信这可能会改变疾病的进程，改善DN患者的生活， 几乎没有治疗选择的疾病。间充质基质/干细胞（MSC）移植提供了希望。 间充质干细胞是一种非胚胎干细胞，具有抗纤维化、抗炎和促血管生成的旁分泌活性 可以改善DN模型的再生。衰老是一种不可逆的细胞周期停滞， 损害邻近细胞功能的促炎分泌表型。因此，衰老细胞增加 DN中的负荷可能实质上损害MSC功能，并成为成功自体移植的障碍。 MSC移植我们的总体目标是表征和优化MSC的功能特性， DN使这些患者能够从未来使用干细胞的临床试验中获益 移植限制MSC功能能力的中心机制，可以通过衰老清除剂治疗， 选择性清除衰老细胞的药物。我们将检查senolytic治疗作为一种潜在的体内 预处理方法以改善干细胞功能。我们的中心假设是脂肪来源的MSC 从DN患者中获得的结果显示衰老增加和功能降低，这可能是 使用清除衰老细胞的药物，在体外和体内都得到改善。首先，我们将比较 从DN患者的MSC到年龄和性别匹配的 对照第二，我们将在体外用衰老清除剂孵育细胞，并评估DN-MSC衰老细胞 清除和功能之后。第三，我们将进行一项试点研究，其中DN患者将接受 将在基线和治疗后14天测量抗衰老药物和MSC衰老和功能。 额外的检查将包括血液和尿液收集肾功能和损伤测量。 所提出的研究探索了一种创新的方法，用于预处理MSC和其有害的 这有助于改善微环境，并有助于开发一种全新的治疗策略来延迟DN进展。

PROJECT SUMMARY/ABSTRACT I am a practicing nephrologist at Mayo Clinic and I will use the acquired skills from my clinical and epidemiology background to bring the proposed basic research investigations into truly translational research for patients with diabetic nephropathy (DN). I have assembled a multidisciplinary team of mentors who will provide guidance, infrastructure, and tools needed for performance of the proposed studies. My immediate career goal is to gain laboratory research skills, clinical trial experience, regenerative medicine fund-of- knowledge, and regenerative medicine coursework. My long-term goal is to become a leader in Regenerative Nephrology and an independent, productive clinician-investigator. My career development plan will provide me with the building blocks needed to transition into independence. I have the strongest support from my division, department, and institution. In the proposed studies, I will explore the feasibility of a novel therapeutic platform that I believe may change the course of disease and improve the lives of patients with DN, a devastating disease with few therapeutic options. Mesenchymal stromal/stem cell (MSC) transplantation offers hope. MSCs are non-embryonic stem cells with anti-fibrotic, anti-inflammatory, and pro-angiogenic paracrine activity that improve regeneration in DN models. Senescence is an irreversible cell cycle arrest, which generates a pro-inflammatory secretory phenotype that impairs neighboring cell function. Hence, increased senescent cell burden in DN may substantially compromise MSC function and become a barrier to successful autologous MSC transplantation. Our overall goal is to characterize and optimize the functional properties of MSC in DN to allow these patients to benefit from future enrollment in clinical trials using stem cell transplantation. A central mechanism limiting MSC functional capacity, may be treatable through senolytic drugs that selectively eliminate senescent cells. We will examine senolytic therapy as a potential in vivo preconditioning method to improve stem cell function. Our central hypothesis is that adipose-derived MSC obtained from DN patients show increased senescence and decreased functionality, which can be ameliorated, both in vitro and in vivo, using drugs that clear senescent cells. First, we will compare cellular senescence and functionality in MSC from DN patients to MSC from age- and gender-matched controls. Second, we will incubate cells with senolytic agents in vitro and assess DN-MSC senescent cell clearance and function thereafter. Third, we will conduct a pilot study wherein DN patients will receive senolytic drugs, and MSC senescence and function will be measured at baseline and 14 days after treatment. Additional examinations will include blood and urine collection for kidney function and injury measurements. The proposed studies explore an innovative approach for preconditioning MSC and their deleterious microenvironment, and aid in developing a completely novel therapeutic strategy to delay DN progression.