A novel approach for treating diabetes using pulsed focused ultrasound and intra-arterial delivery of mesenchymal stem cell based therapies directly into the pancreas

项目总结 1型糖尿病(T1D)的治疗包括终生每日注射胰岛素，以确保严格的代谢控制。 然而，最近的研究表明，糖尿病患者体内仍有残留的功能性β细胞。 胰腺，从而可以恢复和/或再生β细胞数量和功能的治疗性干预 会对这些患者产生深远的影响。保存和再生β细胞的一种有前景的方法是 将以间充质干细胞(MSC)为基础的疗法直接输送到胰腺。MSCs可以释放 免疫调节、血管生成、抗炎、抗凋亡和抗纤维化因子进入其周围 调节免疫系统以及刺激受损组织再生的微环境； 这些旁分泌因子以可溶性形式或在细胞外小泡(EVS)内释放。研究表明， 表明亲本MSCs(即细胞疗法)和MSC来源的EV(即无细胞疗法)都可以 提高胰岛的存活率和功能。不幸的是，基于MSC的治疗方法的临床翻译 糖尿病的治疗一直是次优的，主要是因为大多数骨髓间充质干细胞和电动汽车被困在 肺和网状内皮系统，分别在常规静脉注射(IV)后。因此， 在本提案中，我们将：(I)研究一种使用脉冲聚焦超声(PFUS)的新方法来 轻轻摇动和“启动”胰腺，释放出能够吸引和保留基于MSC的疗法的化学物质。 通过动脉内(IA)注射直接输送到腺体中，以及(Ii)确定哪个来源和 以骨髓间充质干细胞为基础的治疗最适合于糖尿病胰腺的再生和保护。在目标1中，我们将 研究pFUS这一临床可用的技术对胰腺的生物学效应， 单个胰岛和不同来源的MSCs。在初步研究中，我们发现声波可以 它们不仅能刺激胰岛和间充质干细胞，还能诱导趋化因子的表达。 细胞因子、营养因子和细胞黏附分子)；后者将有助于促进 将基于MSC的治疗归位、渗透和保留到糖尿病胰腺内挣扎的胰岛。在……里面 AIMS 2(亲本MSCs)和3(MSC来源的EVS)，我们将评估基于MSC的治疗的效果 不同来源(如骨髓、脂肪组织和脐带)对糖尿病患者再生的影响 当胰腺通过IA注射直接注入腺体时，在用 PFUS。为了实现这些目标，我们开发了一种将治疗药物直接输送到胰腺的技术，通过 它的动脉血液供应，旨在模拟介入放射科医生可以使用 血管内技术。此外，我们将使用一种名为EXTERIC的新型设备来分离来自MSC的电动汽车 为我们的研究提供高纯度和高产量。我们希望共同确定最优参数(即基于MSC 治疗、给药途径和pFUS参数)，可临床翻译用于治疗T1D患者。

PROJECT SUMMARY Treatment for type 1 diabetes (T1D) involves life-long daily injections of insulin to ensure tight metabolic control. However, recent studies have shown that diabetic patients still have residual functional β cells within their pancreas and hence therapeutic interventions that could recover and/or regenerate β cell quantity and function would have a profound impact on these patients. A promising approach to preserve and regenerate β cells is to deliver mesenchymal stem cell (MSC)-based therapies directly to the pancreas. MSCs can release immunomodulatory, angiogenic, anti-inflammatory, anti-apoptotic and anti-fibrotic factors into their surrounding microenvironment to modulate the immune system as well as stimulate the regeneration of damaged tissues; these paracrine factors are released either in a soluble form or within extracellular vesicles (EVs). Studies have shown that both parent MSCs (i.e. a cellular therapy) and MSC-derived EVs (i.e. a cell free therapy) can improve the survival and function of islets. Unfortunately, the clinical translation of MSC-based therapies for the treatment of diabetes has been sub-optimal, predominantly due to majority of MSCs and EVs getting trapped in the lung and reticuloendothelial system, respectively, following conventional intravenous (IV) injection. Hence, in the present proposal, we will: (i) investigate a novel approach using pulsed focused ultrasound (pFUS) to gently shake and “prime” the pancreas to release chemicals which can attract and retain MSC-based therapies that are delivered directly into the gland by intra-arterial (IA) injection and (ii) determine which source and type of MSC-based therapy is best suited to regenerate and protect the diabetic pancreas. In Aim 1, we will investigate the biological effects of pFUS, which is a clinically available technology, on the pancreatic gland, individual pancreatic islets and different sources of MSCs. In pilot studies, we have found that soundwaves can not only stimulate pancreatic islets and MSCs, but they can also induce the expression of chemoattractants (i.e. cytokines, trophic factors, and cell adhesion molecules) in the pancreas; the latter will help to facilitate the homing, permeation and retention of MSC-based therapies to struggling islets within the diabetic pancreas. In Aims 2 (parent MSCs) and 3 (MSC-derived EVs), we will evaluate the effect of MSC-based therapies derived from different sources (i.e. bone marrow, adipose tissue and umbilical cord) on regenerating the diabetic pancreas when they are given directly into the gland via IA injection, before and after “priming” the pancreas with pFUS. To achieve these aims, we developed a technique to deliver therapeutics directly into the pancreas, via its arterial blood supply, which is designed to simulate what Interventional Radiologists can do using endovascular techniques. In addition, we will use a novel device called ExoTIC to isolate MSC-derived EVs with high purity and yield for our studies. Together, we hope to determine the optimal parameters (i.e. MSC-based therapy, delivery route and pFUS parameters) that can be clinically translated for the treatment of T1D patients.