Extracellular vesicles derived from amniotic fluid stem cells normalize glomerular function during progressive kidney disease.

在大多数慢性肾脏疾病(CKD)的进展过程中，足细胞和肾小球内皮细胞(GEC) 都受到了不可逆转的破坏。对这些细胞的损伤或肾小球基底膜成分的改变 膜导致肾小球滤过屏障(GFB)结构和功能的改变。重新- 通过刺激内源性修复机制建立GFB功能可以延缓肾脏疾病 进步。 这项提案中提出的数据显示，细胞外小泡来自人羊水干细胞 (hAFSC-EVS)在我们的CKD，Alport综合征(AS)动物模型中具有体内肾脏保护作用。没有副作用或 免疫反应的刺激发生了。这些EVS可以通过以下方式激活肾小球细胞的修复机制 货物转运。具体地说，它们调节足细胞和GEC中miR-93的水平。MIR-93发挥着关键作用 在慢性肾脏病中的作用，因为其表达水平的变化与肾损害的发展和 具体地说，miR-93在小鼠和人肾小球和尿样中的表达减少。 HEV可以将miR-93恢复到正常水平。我们假设HEV可以重建肾小球功能 通过防止进一步的肾小球损伤，从而最大限度地减少肾脏疾病的进展。使用转基因技术 Alport小鼠、人类来源的EVS和人类AS活检结合创新的空间转录组学 方法，我们将研究EV的作用机制，特别是EV/miR-93轴的能力 对调节肾小球细胞生物学的细胞信号网络进行“重新编程”。我们的目标也是评估 不同AS阶段HEV的疾病改正能力(目标1)。最后，我们打算研究分子 疾病进展过程中肾小球内信号通路的变化，主要集中在GEC和TO 在人类身上验证这些数据作为活组织检查(目标2)。 这项提案的成功完成将为维护 肾小球的结构和功能。重要的是，这一知识很可能适用于其他形式的CKD 并可能有助于发现专门为最小化肾小球而量身定做的新治疗药物 损坏。

During the progression of most chronic kidney diseases (CKD) podocytes and glomerular endothelial cells (GEC) are irreversibly damaged. Injury to these cells or changes within the composition of the glomerular basement membrane lead to alterations of the structure and function of the glomerular filtration barrier (GFB). Re- establishing GFB function by stimulating endogenous repair mechanisms could slow kidney disease progression. Data presented in this proposal show that extracellular vesicles derived from human amniotic fluid stem cells (hAFSC-EVs) are renoprotective in vivo in our animal model of CKD, Alport Syndrome (AS). No side effects or stimulation of an immune response occurred. These EVs can activate repair mechanisms in glomerular cells by cargo transfer. Specifically, they modulate the levels of miR-93 in both podocytes and GEC. miR-93 plays a key role in CKD since changes in its expression level are associated with the development of renal damage and fibrosis; specifically, miR-93 expression is decreased in mouse and human AS glomeruli and urine samples and hEVs can restore miR-93 levels to normal. We hypothesize that hEVs can re-establish glomerular function by preventing further glomerular injury, thus minimizing renal disease progression. Using transgenic Alport mice, EVs of human origin, and human AS biopsies combined with an innovative spatial transcriptomics approach, we will study the EV mechanism of action with specific focus on the ability of the EV/miR-93 axis to “re-program” cellular signaling networks that regulate glomerular cell biology. We also aim to evaluate the disease modifying capability of hEVs at different AS stages (Aim 1). Finally, we aim to study the molecular signaling pathway changes within the glomerulus during disease progression with specific focus on GEC and to validate these data in human AS biopsies (Aim 2). Successful completion of this proposal will provide novel insights into the key factors critical for maintenance of glomerular structure and function. Importantly, this knowledge would likely be applicable to other forms of CKD and possibly will facilitate the discovery of new therapeutic agents tailored specifically to minimize glomerular damage.