Targeted nanodelivery in T1D

由于迄今为止没有有效的治疗方法，正在进行的1型糖尿病（T1 D）流行仍然是一个主要的健康问题。 问题.虽然免疫疗法（IT）对T1 D的治疗有很大的希望，但它们的不足， 严重的毒性、副作用和发病率限制了终身免疫抑制的研究努力 approach.这一缺陷促使调查人员寻找替代方法。针对性 使用聚合物纳米颗粒（NPs）的纳米医学特别有希望增强IT的递送， 治疗T1 D。这种策略可以通过将IT递送给患病的患者来最大限度地减少IT的不良副作用。 组织，在那里它们可以持续释放。在这个多学科项目中，我们的目标是开发一个 创新的靶向纳米递送方法，用于T1 D的IT。虽然在发展方面取得了进展， 新制剂，一种在全身给药后将NP靶向递送至特定组织部位的方法， 行政管理有待发展。自身反应性T细胞的引发和活化发生在 胰腺淋巴结（PLN），其中初始T细胞通过淋巴结（LN）限制的脉管系统进入 称为高内皮小静脉（HEV），并遇到来自胰腺的自身抗原， 树突状细胞活化的T细胞随后运输到胰腺，引起胰岛炎和自身免疫性疾病。 糖尿病值得注意的是，我们已经发现，在糖尿病的发病过程中，HEV也在胰腺中形成。 NOD小鼠。我们的可生物降解聚合物纳米粒通过使用马来酰亚胺-硫醇与MECA 79 IgM抗体包被 化学（MECA 79-MT-NP）; MECA 79结合外周淋巴结地址素（PNAd），一种糖蛋白家族 仅由HEV的内皮细胞表达。在这里，我们第一次展示了有针对性的交付， MECA 79-MT-NP在静脉内施用后对NOD小鼠的PLN和胰腺的作用。我们也 提供支持我们交付平台临床适用性的人体数据。主动定向输送， 这些位点在T1 D中从未实现过。此外，我们的初步数据显示， 我们的MECA 79-MT-NP中的抗CD 3抗体导致更有效地逆转自身免疫性糖尿病， NOD小鼠比用游离抗CD 3治疗的小鼠更好。我们的主要假设是，抗CD 3靶向递送到 胰腺淋巴结（PLN）和胰腺将增加其疗效，并通过减少 全身给药显著。在目标1中，我们将检查和优化稳定性、结合效力和 MECA 79-缀合的NP（MECA 79-NP）的生物分布，其通过利用炔-叠氮化物化学来允许 MECA 79的五聚体形式与NP的连接（MECA 79-AA-NP）。在目标2中，我们将评估 在具有优化性质的MECA 79-NP制剂中包封抗CD 3的临床疗效 目的1逆转NOD小鼠的自身免疫性糖尿病，并阐明其机制， 我们的靶向治疗有效在目标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MECA 79-NP的T1 D患者。这种多学科的合作方法将奠定 为采用创新的、有针对性的T1 D信息技术提供方法奠定基础。

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.