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

项目摘要/摘要 我是梅奥诊所的一名实习肾脏科医生，我将运用从临床和临床中学到的技能 将拟议的基础研究调查纳入真正的翻译研究的流行病学背景 适用于糖尿病肾病(DN)患者。我已经组建了一个多学科的导师团队，他们将 提供执行拟议研究所需的指导、基础设施和工具。我的直系亲属 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，以及再生医学课程。我的长期目标是成为再生领域的领导者 他是一名独立的、富有成效的临床医生兼研究员。我的职业发展计划将为我提供 拥有过渡到独立所需的基石。我得到了我所在部门最强有力的支持， 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糖尿病肾病的病程，改善患者的生活，这是一个毁灭性的 疾病几乎没有治疗选择。间充质干细胞(MSC)移植带来了希望。 MSCs是一种非胚胎干细胞，具有抗纤维化、抗炎和促血管生成旁分泌活性。 改善了DN模型中的再生能力。衰老是一种不可逆转的细胞周期停滞，它会产生 损害邻近细胞功能的促炎性分泌表型。因此，衰老细胞增加 糖尿病肾病的负担可能会严重损害MSC的功能，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 允许这些患者从未来登记使用干细胞的临床试验中受益 移植。一种限制MSC功能的中枢机制，可通过感觉神经溶解来治疗 选择性地消除衰老细胞的药物。我们将在活体内检测衰老治疗的可能性。 改善干细胞功能的预适应方法。我们的中心假设是脂肪来源的间充质干细胞 从糖尿病肾病患者身上获得的数据显示出衰老加剧和功能下降，这可能是 使用清除衰老细胞的药物，在体外和体内都有所改善。首先，我们将比较 从糖尿病肾病患者到年龄性别匹配的骨髓间充质干细胞的细胞衰老和功能 控制。其次，我们将在体外用促衰老物质孵育细胞，并评估dN-MSC衰老细胞。 清关及其后的运作。第三，我们将进行一项试点研究，糖尿病肾病患者将接受 在治疗前和治疗后14天分别检测抗衰老药物和MSC的衰老和功能。 其他检查将包括采血和尿液进行肾功能和损伤测量。 拟议的研究为MSC的预适应及其危害探索了一种创新的方法 微环境，并帮助开发一种全新的治疗策略，以延缓糖尿病肾病的进展。

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.