Mesenchymal stem cell senescence in diabetic nephropathy

 描述（由申请人提供）：我是梅奥诊所的一名执业肾脏病专家和慢性肾脏病 (CKD) 诊所的联席主任，也是美国肾脏病学会透析咨询小组的成员。 My research to date has examined patient-important outcomes in dialysis, hypertension, and kidney transplant populations.我将利用我从临床和流行病学背景中获得的技能，将拟议的基础研究调查转化为糖尿病肾病（DN）患者的真正转化研究。在我的 为了改善 CKD 患者的生活，我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。我的近期职业目标是获得实验室研究技能、临床试验经验、再生医学知识基金以及以再生医学为重点的临床和转化科学硕士学位。 My long-term goal is to become a leader in Regenerative Nephrology and an independent, productive clinician-investigator.我在 K23 期间的职业发展计划包括实验室轮换、再生医学中心的现场和异地实习、个性化课程作业、出版物、协作网络建设、在国家会议上通过摘要演示进行接触，以及临床试验方案设计和实施的经验。 These endeavors will provide me with the building blocks needed to transition into independence. I have the strongest support from my division, department, and institution.在拟议的研究中，我将探索一种新型治疗平台的可行性，我相信该平台可能会改变疾病进程并改善 DN 患者的生活，DN 是一种治疗选择很少的毁灭性疾病。 DN is the most prevalent cause of CKD and resistant to most interventions aimed at preventing progression. However, recent advances in regenerative medicine of adipose tissue-derived mesenchymal stromal/stem cell (MSC) transplantation offer hope. MSC 是非胚胎干细胞，具有抗纤维化、抗炎和促血管生成旁分泌活性，可改善 DN 模型的再生。 However, patient-specific factors such as aging, obesity, uremia, and diabetes may decrease cellular function by inducing cellular senescence. Senescence is an irreversible cell cycle arrest, which generates a pro-inflammatory secretory phenotype that impairs neighboring cell function.因此，DN 中衰老细胞负担的增加可能会严重损害 MSC 功能，并成为成功自体 MSC 移植的障碍。我们的总体目标是表征和优化 DN 中 MSC 的功能特性，使这些患者能够从未来参加干细胞移植临床试验中受益。我们令人兴奋的新数据表明，细胞衰老是限制间充质干细胞功能能力的一种核心机制，可以通过选择性消除衰老细胞的衰老药物来治疗。 In accordance, we showed that eradicating senescent cells improves stem cell function in animal models. Therefore, we plan to examine senolytic therapy as a potential in vivo preconditioning method to improve stem cell function.我们所提出的研究的中心假设是，从 DN 患者中获得的脂肪组织来源的 MSC 显示出衰老增加和功能下降，这可以通过使用清除衰老细胞的药物在体外和体内得到改善。 This hypothesis will be pursued in 3 specific aims.首先，我们将比较来自 DN [估计肾小球滤过率 (eGFR) 15-60 mL/min/1.73m2] 患者的脂肪组织来源的 MSC 与来自年龄和性别匹配对照的 MSC 的细胞衰老和功能。其次，为了确定 DN-MSC 功能障碍的可逆性，我们将在体外将细胞与衰老细胞一起孵育，然后评估 DN-MSC 衰老细胞的清除和功能。第三，为了检查体内衰老药物对 DN-MSC 功能的影响，我们将进行一项试点研究，其中 DN 患者（eGFR 15-45 mL/min/1.73m2）将接受衰老药物，并在基线和治疗后 14 天测量 MSC 衰老和功能，并与未治疗的对照进行比较。 For each of these studies, DN patients will undergo 1-2 abdominal fat biopsies for MSC harvesting. Additional examinations will include blood and urine collection for diabetes, kidney function, and CKD-related measurements.意义：开发一种安全有效的疗法来延缓 DN 进展可以降低与透析相关的发病率，为经常推迟肾移植的人群提供更好的治疗选择，并节省大量成本。这些新颖的研究将增进人们对细胞衰老对 MSC 影响的认识，并有助于制定预筛选方案，以优化使用自体 MSC 移植治疗 DN 的试验的招募。此外，拟议的研究探索了一种预处理 MSC 及其有害微环境的创新方法，并有助于开发一种全新的治疗策略来延缓 DN 的进展。 As a nephrologist, I am passionate about improving the lives of patients with CKD through innovative research.我相信，我的决心、临床和研究背景、最佳和支持性的机构环境、出色的多学科指导和咨询团队，再加上 K23 的受保护时间和资金，将推动我实现成为一名独立的临床医生研究员并将干细胞疗法带给 DN 患者的目标。

 DESCRIPTION (provided by applicant): I am a practicing nephrologist and Co-Director of the Chronic Kidney Disease (CKD) clinic at Mayo Clinic, and member of the American Society of Nephrology Dialysis Advisory Group. My research to date has examined patient-important outcomes in dialysis, hypertension, and kidney transplant populations. 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). In my quest to improve the lives of patients with CKD, 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 a Master's degree in Clinical and Translational Science with a focus on regenerative medicine. My long-term goal is to become a leader in Regenerative Nephrology and an independent, productive clinician-investigator. My career development plan during the K23 period entails laboratory rotations, on-site and off-site clerkships in centers for regenerative medicine, individualized coursework, publications, collaborative network building, exposure through abstract presentations at national conferences, and experience with clinical trial protocol design and implementation. These endeavors 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. DN is the most prevalent cause of CKD and resistant to most interventions aimed at preventing progression. However, recent advances in regenerative medicine of adipose tissue-derived mesenchymal stromal/stem cell (MSC) transplantation offer hope. MSCs are non-embryonic stem cells with anti-fibrotic, anti-inflammatory, and pro-angiogenic paracrine activity that improve regeneration in DN models. However, patient-specific factors such as aging, obesity, uremia, and diabetes may decrease cellular function by inducing cellular senescence. 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. Our exciting new data reveal that cellular senescence, a central mechanism limiting MSC functional capacity, may be treatable through senolytic drugs that selectively eliminate senescent cells. In accordance, we showed that eradicating senescent cells improves stem cell function in animal models. Therefore, we plan to examine senolytic therapy as a potential in vivo preconditioning method to improve stem cell function. Our central hypothesis underlying the proposed studies is that adipose tissue-derived MSC obtained from patients with DN show increased senescence and decreased functionality, which can be ameliorated, both in vitro and in vivo, using drugs that clear senescent cells. This hypothesis will be pursued in 3 specific aims. First, we will compare cellular senescence and functionality in adipose tissue-derived MSC from patients with DN [estimated glomerular filtration rate (eGFR) 15-60 mL/min/1.73m2] to MSC from age- and gender-matched controls. Second, to determine the reversibility of DN-MSC dysfunction, we will incubate cells with senolytic agents in vitro and assess DN-MSC senescent cell clearance and function thereafter. Third, to examine the effect of senolytic agents on DN-MSC function in vivo, we will conduct a pilot study wherein DN patients (eGFR 15-45 mL/min/1.73m2) will receive senolytic drugs, and MSC senescence and function will be measured at baseline and 14 days after treatment with comparison to untreated controls. For each of these studies, DN patients will undergo 1-2 abdominal fat biopsies for MSC harvesting. Additional examinations will include blood and urine collection for diabetes, kidney function, and CKD-related measurements. Significance: Developing a safe and effective therapy to delay DN progression could reduce morbidity associated with dialysis, offer a better treatment option to a population often deferred for kidney transplantation, and produce extensive cost savings. These novel studies will advance the knowledge of the effects of cellular senescence on MSC, and help develop pre-screening protocols to optimize enrollment in trials using autologous MSC transplantation for DN. Furthermore, 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 the progression of DN. As a nephrologist, I am passionate about improving the lives of patients with CKD through innovative research. I believe my determination, clinical and research background, optimal and supportive institutional environment, and outstanding multidisciplinary mentoring and advisory team in combination with the protected time and funding from the K23 will propel me toward my goals of becoming an independent clinician-investigator and bringing stem cell therapy to patients with DN.