Exosomes from miR-primed endothelial progenitor cells for treating ischemic stroke

项目摘要 缺血性中风(IS)，由于脑血管闭塞而导致的脑血流丧失，仍然是 世界上最严重的健康问题。目前对中风的治疗方法，如溶栓剂、PTAS、 神经保护性药物已经被证明是有帮助的；然而，它们也受到治疗时间的限制。 窗口，再中风的风险和低有效率。寻找一种理想的保护大脑的治疗策略 因此，急性损伤和促进血管生成和神经再生对于神经功能恢复是势在必行的 需要的。内皮祖细胞(EPC)具有分化为成熟内皮细胞(ECs)的能力， 已知参与血管生成和神经生成。内皮祖细胞移植显示出良好的效果 因为这是治疗。新出现的证据表明，干细胞释放的exs可能用于治疗IS。EXS是 内体起源的小膜小泡通过转运miRNAs(MiRs)介导细胞间的通讯。 与干细胞相比，干细胞exs具有优势，因为它缺乏自我增殖能力，更容易存储和 比细胞更能传递。在这个项目中，我们的目标是使用epc释放的exs(epc-exs)作为IS治疗的一种新途径。 因为它们选择性地携带浓缩的miR-126。我们发现EPC-exs保护ECs不受 缺氧/复氧(H/R)引起的损伤和功能障碍主要通过其携带的miR-126； EPC-exs能保护脑血流量，缩小脑梗塞体积。有趣的是，反- 细胞凋亡/抗氧化应激miR-210已被发现在中风患者中减少。我们发现 MiR-210诱导的EPC-exs(EPC-EXsmiR210)比EPC-exs更有效地保护内皮细胞免受缺氧-再灌注损伤。 致伤。综上所述，通过结合EPC(干细胞)、Exs(新玩家)的有益效果 脑微环境)和MIR(多信号通路靶点)，我们认为EPC-EXsmiR210可以 作为IS的一种新的治疗途径。设计了三个目标来验证我们的假设：EPC-EXsmiR210 通过保护脑细胞免受急性缺血性损伤而增强治疗IS的效果 血管生成/神经再生通过EPC-exs、miR-126和miR-210促进神经功能恢复 下游途径：VEGF/VEGFR2/PI3K/Akt、PIK3R2/PI3K/Caspase和BDNF/TrkB/PI3K/Akt。目标1： 确定EPC-EXsmiR210是否增强了对内皮细胞和神经元的保护作用 通过靶向多个途径造成损伤/功能障碍。目标2：确定EPC-EXsmiR210是否增强 靶向多途径对IS急性脑损伤的保护作用。目标3：确定 EPC-EXsmiR210在促进神经血管再生和功能恢复方面是否有增强作用 就是瞄准多条路径。 。

Project Summary Ischemic stroke (IS), loss of cerebral blood flow due to occlusion of cerebral blood vessels, remains one of the most severe health problems in the world. Current treatments for stroke, such as thrombolytic agents, PTAS, and neuroprotective drugs have shown to be helpful; however, they are also limited by a narrow therapeutic time window, risk of re-stroke and low effectives. Looking for an ideal therapeutic strategy that protect brain from acute injury and promote angiogenesis and neurogenesis for neurological functional recovery, is thus imperative needed. Endothelial progenitor cells (EPCs) with the ability to differentiate into mature endothelial cells (ECs), are known to participate in angiogenesis and neurogenesis. EPC transplantation has shown promising results for IS therapy. Emerging evidence suggests that stem cell released EXs could be used for treating IS. EXs are endosomal origin small-membrane vesicles mediating cell-cell communication by transferring miRNAs (miRs). Stem cell EXs hold advantages over stem cells because of lacking self-proliferation and are easier to store and deliver than cells. In this project, we aim to use EPC-released EXs (EPC-EXs) as a novel avenue for IS therapy as they selectively carry enriched miR-126. We have found that EPC-EXs protect ECs from hypoxia/reoxygenation (H/R)-induced injury and dysfunction mainly through their carried miR-126; transfusion of EPC-EXs could preserve cerebral blood flow (CBF) and decrease infarct volume after IS. Of interest, the anti- apoptosis/anti-oxidative stress miR-210 has been found to be decreased in the stroke patients. We discovered that miR-210 primed EPC-EXs (EPC-EXsmiR210) are more effective than EPC-EXs in protecting ECs from H/R- induced injury. Taken together, by combining the beneficial effects of EPCs (stem cells), EXs (novel player of brain microenvironment), and miRs (multiple signaling pathway targets), we believe that EPC-EXsmiR210 could serve as a novel therapeutic avenue for IS. Three aims are designed to test our hypothesis that EPC-EXsmiR210 have enhanced therapeutic effects on IS by protecting brain cells from acute ischemic injury and promoting angiogenesis/neurogenesis for neurological functional recovery through EPC-EXs, miR-126 and miR-210 downstream pathways, VEGF/VEGFR2/PI3K/Akt, PIK3R2/PI3K/Caspase and BDNF/TrkB/PI3K/Akt. Aim 1: Determine whether EPC-EXsmiR210 have enhanced effects on protecting ECs and neurons from H/R-induced injury/dysfunction by targeting multiple pathways. Aim 2: Determine whether EPC-EXsmiR210 exert enhanced effects on protecting brain from IS-induced acute damage by targeting multiple pathways. Aim 3: Determine whether EPC-EXsmiR210 exert enhanced effects on promoting neurovascular genesis and functional recovery after IS by targeting multiple pathways. .