Targeted nanodelivery in T1D

由于到目前为止还没有有效的治疗方法，正在进行的1型糖尿病(T1D)流行仍然是一个主要的健康问题 有问题。虽然免疫疗法(ITS)为治疗T1D带来了巨大的希望，但它们的不足之处， 严重的毒副作用和发病率限制了终生免疫抑制的研究努力。 接近。这一缺陷促使调查人员寻找替代方法。目标明确 使用聚合物纳米颗粒(NPs)的纳米医学具有特别的前景，可以增强其对 治疗T1D。这种策略可以最大限度地减少ITS的不良副作用，将其传递给患者 组织，在那里它们可以进行持续的释放。在这个多学科的项目中，我们的目标是开发一种 创新的、靶向的T1D ITS纳米递送方法。尽管在发展方面取得了进展 新的配方，一种将NPs靶向输送到特定组织部位的方法 行政管理有待发展。自身反应性T细胞的启动和激活发生在 胰腺淋巴结(PLN)，原始T细胞通过受LN限制的血管系统进入 称为高内皮微静脉(HEV)，并遇到来自胰腺的自身抗原 树突状细胞。激活的T细胞随后进入胰腺，导致胰岛炎症和自身免疫 糖尿病。值得注意的是，我们发现在#年糖尿病发作期间，胰腺中也形成了HEV。 点头老鼠。利用马来酰亚胺-硫醇将MECA79-IgM抗体包覆在可生物降解的聚合物纳米粒上 化学(MECA79-MT-NP)；MECA79结合外周节点寻址蛋白(PNAD)，一个糖蛋白家族 仅在HEV的内皮细胞中表达。在这里，我们首次展示了有针对性地交付 静脉注射MECA79-MT-NPs对NOD小鼠PLN和胰腺的影响我们也 提供人力数据，支持我们的交付平台的临床适用性。主动定向投递至 这些站点在T1D中从未实现过。此外，我们的初步数据显示，IT的封装 MECA79-MT-NPs中的抗CD3抗体可更有效地逆转小鼠自身免疫性糖尿病 NOD小鼠给予游离抗CD3抗体治疗。我们的主要假设是将抗CD3的靶向递送到 胰腺淋巴结节(PLN)和胰腺组织将通过减少 全身性用药显著。在目标1中，我们将检查和优化稳定性、结合效率和 利用炔叠氮化物化学方法研究MECA79-NPs的生物分布 将MECA79的五聚体连接到NP上(MECA79-AA-NP)。在目标2中，我们将评估 优化性能的MECA79-NP制剂中包裹抗CD3抗体的临床疗效 目的1逆转NOD小鼠自身免疫性糖尿病，并阐明其机制。 我们的靶向治疗奏效了。在目标3中，我们计划测试与人的PLN和胰腺的结合能力 我们优化的MECA79-NPs的T1D患者。这种多学科、协作的方法将为 为为T1D引入创新的、有针对性的ITS交付方法奠定了基础。

With no effective therapy to date, the ongoing Type 1 diabetes (T1D) epidemic continues to be a major health problem. While immune therapeutics (ITs) hold great promise for the treatment of T1D, their inadequacy, serious toxicity, side effects, and morbidity have limited research efforts in the lifelong immunosuppression approach. This shortcoming has prompted investigators to search for alternative approaches. Targeted nanomedicine using polymeric nanoparticles (NPs) holds particular promise to enhance the delivery of ITs to treat T1D. This strategy can minimize the undesirable side effects of ITs by delivering them to diseased tissues, where they can undergo sustained release. In this multidisciplinary project, we aim to develop an innovative, targeted nanodelivery method for ITs for T1D. Although progress has been made in developing new formulations, a method of targeted delivery of NPs to specific tissue sites following systemic administration remains to be developed. The priming and activation of autoreactive T cells occurs in the pancreatic lymph nodes (PLNs), where naive T cells enter through lymph node (LN)-restricted vasculature known as high endothelial venules (HEVs) and encounter autoantigens from the pancreas presented by dendritic cells. Activated T cells traffic subsequently to the pancreas, causing insulitis and autoimmune diabetes. Notably, we have found that HEVs are also formed in the pancreas during the onset of diabetes in NOD mice. Our biodegradable polymeric NPs are coated with MECA79 IgM antibody by using maleimide-thiol chemistry (MECA79-MT-NP); MECA79 binds to peripheral node addressin (PNAd), a glycoprotein family expressed only by endothelial cells of the HEV. Here, we demonstrate for the first time the targeted delivery of MECA79-MT-NPs to the PLNs and pancreata of NOD mice following intravenous administration. We also provide human data that supports the clinical applicability of our delivery platform. Active targeted delivery to these sites has never been achieved in T1D. Moreover, our preliminary data shows that encapsulation of the IT anti-CD3 antibody inside our MECA79-MT-NPs results in more effective reversal of autoimmune diabetes in NOD mice than treatment with free anti-CD3. Our main hypothesis is that targeted delivery of anti-CD3 to the pancreatic lymph nodes (PLNs) and pancreata will increase its efficacy and decrease toxicity by reducing systemic dosing significantly. In Aim 1, we will examine and optimize the stability, binding efficacy, and biodistribution of MECA79-conjugated NPs (MECA79-NPs) by utilizing alkyne-azide chemistry to permit the attachment of the pentameric form of MECA79 to the NP (MECA79-AA-NP). In Aim 2, we will assess the clinical efficacy of encapsulating anti-CD3 inside the formulation of MECA79-NP with the optimized properties from Aim 1 in the reversal of autoimmune diabetes in NOD mice as well as elucidate the mechanisms by which our targeted therapy works. In Aim 3, we plan to test the binding capacity to the PLNs and pancreata of human T1D patients of our optimized MECA79-NPs. This multidisciplinary, collaborative approach will lay the groundwork for the introduction of an innovative, targeted delivery method of ITs for T1D.