Hypoxic preconditioning for the improvement of the regenerative capacity of adipose-derived mesenchymal stem cells and its application in chronic kidney disease due to hypertensive nephropathy

项目总结/摘要 高血压肾病（HN）是慢性肾病（CKD）的第二大原因， 采取有效措施，防止病情发展。然而，再生医学的最新进展， 脂肪组织来源的间充质基质/干细胞（AD-MSC）移植为这些人提供了希望。 患者MSC具有抗纤维化、抗炎和促血管生成的旁分泌活性， 在一些肾损伤模型中再生。然而，暴露于患者特异性因素，如衰老和 HN中CKD的尿毒症环境可能对MSC的表观遗传和转录调节， 将它们的表型改变为对邻近细胞有害的表型。因此，HN中细胞损伤的增加可能 实质上损害MSC功能并成为成功自体MSC移植的障碍。 我们的中心假设是，从HN患者中获得的MSC表现出受损的 功能和血管生成，这可以通过低氧预处理（HPC）进行修改。这一假设将 我们将在体外和体内以三个具体目标进行解决：1）我们将比较来自 HN患者（eGFR 15-60 mL/min/1.73m2）与年龄和性别匹配的高血压患者的MSC 2）为了确定HN-MSC功能障碍的可逆性，我们将来自这些细胞的细胞进行实验， 3）在体外检测MSC功能对HPC的影响，然后评估MSC功能;和 体内，我们将在猪肾血管性高血压模型中进行实验研究， HN的病理生理学，诱导肾动脉狭窄后6周。这些猪，以及正常的 年龄匹配的猪将用HPC-MSC、未处理的MSC或媒介物处理。评估HPC对 四周后，将使用MSC研究MSC效力、肾血流动力学、功能和氧合。 复杂的体内CT和MR成像技术，以及离体研究的肾损伤和血管分布。的 由于开发了一种安全有效的治疗方法来延迟HN，因此所提出的研究的意义很高。 进展可以降低与透析相关的发病率，为人群提供更好的治疗选择， 通常推迟肾移植，并节省大量费用。这些新的研究将 推进HPC对MSC及其有害微环境的影响的知识， 开发一种全新的治疗策略，以延迟支持NIDDK的HN进展 使命。拟议的项目涉及如何表征和改善MSC的功能特性， HN允许这些患者从未来使用干细胞移植的临床试验中获益。的 马约诊所的特殊资源和机构支持，杰出的多学科指导团队， 和拟议的职业发展活动将使候选人实现她的长期目标，成为 他是一位独立的研究者，也是全国公认的肾脏再生医学的领导者。

PROJECT SUMMARY/ABSTRACT Hypertensive Nephropathy (HN) is the second leading cause of chronic kidney disease (CKD) resistant to effective intervention to prevent progression. However, recent advances in regenerative medicine with the use of adipose tissue-derived mesenchymal stromal/stem cell (AD-MSC) transplantation offer hope for these patients. The MSCs have anti-fibrotic, anti-inflammatory, and pro-angiogenic paracrine activities that improve regeneration in some kidney injury models. However, exposure to patient-specific factors, such as aging and the uremic milieu of CKD in HN, may exert epigenetic and transcriptional modulation of MSC, potentially modifying their phenotype to one noxious to neighboring cells. Hence, increased cellular damage in HN may substantially compromise MSC function and become a barrier to successful autologous MSC transplantation. Our central hypothesis underlying this proposal is that MSCs obtained from patients with HN show impaired functionality and angiogenesis, which can be modified with hypoxic preconditioning (HPC). This hypothesis will be addressed both in vitro and in vivo in three specific aims: 1) we will compare functionality of MSCs from patients with HN (eGFR 15-60 mL/min/1.73m2) to MSC from age- and sex-matched patients with hypertension and healthy controls; 2) to determine the reversibility of HN-MSC dysfunction, we will subject cells from these cohorts to HPC in vitro and assess MSC function thereafter; and 3) to examine the effect of MSC function in vivo, we will conduct an experimental study in a porcine renovascular hypertension model that recapitulates the pathophysiology of HN, six-weeks after induction of renal artery stenosis. These pigs, as well as normal age-matched pigs, will be treated with HPC-MSC, untreated-MSC, or vehicle. To assess the impact of HPC on MSC potency, renal hemodynamics, function, and oxygenation will be studied after four-weeks, using sophisticated in vivo CT and MR imaging techniques, and renal injury and vascularity studied ex vivo. The significance of the proposed studies is high, since developing a safe and effective therapy to delay HN 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 HPC on MSC and their deleterious microenvironment, aid in developing a completely novel therapeutic strategy to delay the progression of HN supporting the NIDDK mission. The proposed project addresses how characterize and improve the functional properties of MSC in HN to allow these patients to benefit from future enrollment in clinical trials using stem cell transplantation. The exceptional resources and institutional support at Mayo Clinic, outstanding multi-disciplinary mentorship team, and proposed career development activities will allow the candidate to achieve her long-term goal of becoming an independent investigator and a nationally recognized leader in regenerative medicine in nephrology.