Treating Kidney Injury by Modulating Heat Shock Proteins Using Soundwaves Combined with Mesenchymal Stem Cells and Their Extracellular Vesicles

项目摘要 急性肾损伤（阿基）的特征在于肾滤过率的快速下降，并且如果不治疗可导致 肾衰竭据估计，每年有60万例新的阿基病例， 药物选择和支持性血流动力学优化，目前没有批准的治疗。一个 阻止并可能逆转阿基进展的创新方法是使用间充质干细胞 (MSC)基础疗法。MSC充当“移动的药库”，通过以下途径保护和再生受损细胞： 抗炎、血管生成、免疫调节、抗纤维化和抗凋亡因子，其被释放 以可溶形式或在细胞外囊泡（EV）内。在最近的工作中，我们已经表明，父母双方 MSC（即细胞疗法）和MSC衍生的EV（即无细胞疗法）可以改善动物存活率， 通过调节热休克蛋白（HSP）通路，改善阿基后的肾功能。虽然基于MSC 治疗已经在阿基的临床前模型中显示出相当大的前景，它们的临床转化已经 次优一个主要原因是，这些疗法不能达到受伤的肾脏时，给予 通过常规静脉内（IV）施用，将大部分亲本MSC捕获在患者体内。 肺和MSC衍生的EV被困在网状内皮系统（RES）中。因此，我们将研究如果 我们可以通过动脉内（IA）注射将MSC直接递送到受损的肾脏中来优化MSC疗法。 我们还将研究一种使用声波的新技术的效果，称为脉冲聚焦 超声波（pFUS）对MSC治疗和损伤的肾脏的作用，因为它可以（i）刺激MSC，（ii） 通过创建“分子邮政编码”来调节肾脏微环境，以促进MSC归巢， 保留，和/或（iii）独立地刺激HSP途径以促进肾再生。集中 超声波是FDA批准的技术，目前在临床上使用，尽管不是用于该适应症。我们 将使用阿基和GMP级人骨髓来源的MSC（BM-MSC）的小鼠模型， 还衍生EV-鉴于用于我们所有实验的基于MSC的疗法将衍生自单一来源， 这将允许实验的重复性，并使我们的结果能够在我们所有的目标之间进行比较。在 目的1，我们将研究声波如何影响骨髓间充质干细胞，以及急性损伤的肾脏，在一个特定的时间点， 分子水平。在目的2和3中，我们将比较亲本BM-MSC或BM-MSC的IV施用。 衍生EV，局部IA直接给药至肾脏-我们实验室开发的技术 它模仿了介入放射科医生可以使用微创图像引导在人体中进行的操作， 血管内技术。接下来，我们将确定基于MSC的疗法如何调节分子途径， 参与肾脏再生，特别关注HSP。最后，我们将研究pFUS是否可以 优化阿基背景下MSC疗法的再生能力。

PROJECT SUMMARY Acute kidney injury (AKI) is characterized by a rapid decline in kidney filtration, and if left untreated can lead to kidney failure. It is estimated there are 600,000 new cases of AKI each year, however, beyond careful medication selection and supportive hemodynamic optimization, there is currently no approved therapy. An innovative approach to halt, and possibly reverse, the progression of AKI is to use mesenchymal stem cell (MSC) based therapies. MSCs act as a “mobile drug store” to protect and regenerate damaged cells through anti-inflammatory, angiogenic, immunomodulatory, anti-fibrotic and anti-apoptotic factors, which are released either in a soluble form or within extracellular vesicles (EVs). In recent work, we have shown that both parent MSCs (i.e. a cellular therapy) and MSC-derived EVs (i.e. a cell-free therapy) can improve animal survival and kidney function following AKI, by modulating the heat shock protein (HSP) pathway. Although MSC based therapies have shown considerable promise in preclinical models of AKI, their clinical translation has been suboptimal. A major reason for this is that these therapies cannot reach the injured kidney when given to patients by conventional intravenous (IV) administration, with majority of parent MSCs getting trapped in the lungs and MSC-derived EVs getting trapped in the reticuloendothelial system (RES). Hence, we will examine if we can optimize MSC therapies by delivering them directly into the injured kidney by intra-arterial (IA) injection. We will also investigate the effect of a novel technology which uses sound waves, called pulsed focused ultrasound (pFUS), on both MSC therapies and the injured kidney given that it can (i) stimulate MSCs, (ii) modulate the kidney microenvironment by creating a “molecular zip-code to facilitate MSC homing and retention, and/or (iii) independently stimulate the HSP pathway to facilitate kidney regeneration. Focused ultrasound is an FDA-approved technology which is currently used clinically, albeit not for this indication. We will use a mouse model of AKI and GMP grade human bone marrow derived MSCs (BM-MSCs) from which we also derive EVs – given that MSC based therapies for all our experiments will be derived from a single source, this will allow for experimental reproducibility and enable our results to be compared across all our aims. In Aim 1, we will examine how sound waves affect BM-MSCs, as well as the acutely injured kidney, at a molecular level. In Aims 2 and 3, we will compare IV administration of either parent BM-MSCs or BM-MSC derived EVs, with locoregional IA administration directly into the kidneys - a technique developed by our Lab that mimics what Interventional Radiologists can perform in humans using minimally invasive image guided endovascular techniques. Next, we will determine how MSC based therapies modulate the molecular pathways involved in kidney regeneration, focusing specifically on HSPs. Finally, we will examine whether pFUS can optimize the regenerative capacity of MSC therapies in the context of AKI.