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

项目总结 急性肾损伤(AKI)的特点是肾滤过功能迅速下降，如果不治疗可能会导致 肾衰竭。然而，据估计，每年有60万新的AKI病例，尽管如此 在药物选择和支持性血流动力学优化方面，目前还没有批准的治疗方法。一个 阻止甚至可能逆转AKI进展的创新方法是使用间充质干细胞 (MSC)为基础的疗法。骨髓间充质干细胞扮演着“移动药店”的角色，通过 抗炎、血管生成、免疫调节、抗纤维化和抗凋亡因子，它们被释放 以可溶形式或在细胞外小泡(EVS)内。在最近的研究中，我们已经证明了父母双方 MSCs(即细胞疗法)和MSC衍生的EV(即无细胞疗法)可以提高动物的存活率和 AKI后的肾功能，通过调节热休克蛋白(HSP)途径。尽管基于MSC 在AKI的临床前模型中，治疗方法显示出相当大的前景，它们的临床翻译已经 不太理想。一个主要原因是，这些治疗方法在接受治疗时不能到达受损的肾脏。 患者通过常规静脉(IV)给药，大多数父母MSCs被困在 肺和MSC来源的电动汽车被困在网状内皮系统(RES)。因此，我们将研究是否 我们可以通过动脉内注射(IA)将MSC直接输送到受损的肾脏来优化MSC治疗。 我们还将研究一种名为脉冲聚焦的新技术的效果。 超声(PFUS)，对骨髓间充质干细胞治疗和损伤的肾脏，因为它可以(I)刺激MSCs，(Ii) 通过创建分子邮政编码来调节肾脏微环境，以促进MSC的归巢和 和/或(Iii)独立地刺激热休克蛋白途径以促进肾脏再生。聚焦 超声是FDA批准的一项技术，目前正在临床上使用，尽管不是用于这一适应症。我们 将使用AKI和GMP级人骨髓来源的MSCs(BM-MSCs)的小鼠模型 也衍生EVS-鉴于我们所有实验的基于MSC的疗法都将来自单一来源， 这将允许实验的重复性，并使我们的结果能够在所有目标上进行比较。在……里面 目标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。 分子水平。在目标2和3中，我们将比较父母BM-MSCs或BM-MSC的静脉注射 衍生EVS，局部区域IA直接进入肾脏--这是我们实验室开发的一项技术 这模仿了介入放射科医生使用微创图像引导在人类身上所能做的事情 血管内技术。接下来，我们将确定基于间充质干细胞的疗法是如何调节分子通路的。 参与肾脏再生，特别关注热休克蛋白。最后，我们将检查pFUS是否可以 在AKI的背景下优化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.