Caveolae-Targeted Cisplatin Immunoconjugates for Effective Lung Cancer Therapy

摘要 内皮细胞表面的小凹可以选择性、快速和主动地将靶向抗体泵出 即使当它们与高分子质量结合时，也会进入体内的血液和底层组织 货物。基于这一令人兴奋的最新发现，我们还发现小穴抽水系统能够 前所未有的肿瘤特异性靶向和穿透。综上所述，我们的证据有力地支持了 靶向肿瘤小窝是一种新的策略，它提供了一个通过血管输送治疗药物的门户 内皮细胞屏障并直接进入肿瘤。小窝可以快速地泵送带有附着的靶向抗体 货物通过内皮细胞达到实体肿瘤内的浓度大大超过最大值 血液中的浓度。在这个项目中，我们将重点测试抗体的治疗效用- 顺铂-葡聚糖免疫结合物靶向肿瘤内皮腔内截短型Annexin A1 并有效地穿透实体肿瘤。这些数据将为我们的创新交付提供原则证明 肺癌治疗的策略和走向临床的转移--评估小凹的情况 免疫靶向和泵入肿瘤可以增强化疗的治疗效果。我们将设计， 顺铂-羧甲基葡聚糖-AnnA1抗体的合成及体内评价 结合物将创造新的肿瘤小窝靶向疗法，以增强肺癌的递送和疗效 使用我们先进的动物模型的模型。我们假设通过注射带有顺铂的抗体- CM葡聚糖进入肿瘤，小窝可以迅速和特异性地在肿瘤内浓缩治疗药物和 增强肿瘤杀伤力，并显著降低毒性。该项目的具体目标是：目标1-- 表征针对EC小凹的顺铂免疫结合物在肿瘤中的体内递送，并目的2- 评价EC小凹靶向顺铂免疫结合物的治疗效果。这个项目将利用我们的 除自发性乳腺癌外的新的活体显微镜(IVM)肿瘤模型系统 转基因小鼠。IVM允许直接可视化靶向和内皮处理以及 间质和肿瘤细胞反应，所有这些都可以量化，以帮助提供治疗的新见解 肿瘤的机制。靶向小凹打开了一条穿过限制性血管内皮细胞的特定通道 障碍。它可以提供更接近理想目标的强化交付手段，以实现更多 有效的治疗方法。我们已经证明了AnnA1在人类的血管系统和小窝中表达。 原发和转移性肺肿瘤。总体而言，我们提议的项目可能会创造一个范式转变，从 被动的血管给药，极大地限制了人类的药物疗效，提供了第一个原型 针对小窝的抗癌疗法，以供将来转化为临床使用。

Abstract Caveolae at the endothelial cells surface can selectively, rapidly, and actively pump targeted antibodies out of the bloodstream and into underlying tissue in vivo, even when they are conjugated to high-molecular weight cargo. Based on this exciting recent discovery, we also found that caveolae pumping system enables unprecedented tumor-specific targeting and penetration. Taken together, our evidence strongly supports that targeting tumor caveolae is a new strategy providing a portal to deliver therapeutic agents across the vascular endothelial cell barrier and directly into tumors. Caveolae can rapidly pump a targeted antibody with the attached cargo across the endothelial cells to reach concentrations inside solid tumors that greatly exceed maximum concentrations in the blood. In this project we will focus on testing of the therapeutic utility of the antibody- cisplatin-CMdextran immunoconjugates to target truncated form of Annexin A1 in tumor endothelial caveolae and to effectively penetrate solid tumors. The data will provide proof-of-principle of our innovative delivery strategy for therapy of lung cancer and moves toward clinical translation by assessing how well caveolae immunotargeting and pumping into tumors can enhance the therapeutic impact of chemotherapy. We will design, synthesize and asses in vivo the efficacy of cisplatin-carboxymethyl dextran (CMdextran)- AnnA1 antibody conjugate to create new tumor caveolae-targeted therapeutics for enhanced delivery and efficacy in lung cancer models using our advanced animal models. We hypothesize that by pumping antibodies armed with cisplatin- CMdextran into tumors, caveolae can rapidly and specifically concentrate therapeutic agents inside tumors and enhance tumor destruction with significantly reduced toxicity. The Specific Aims of this project are: Aim 1 - to characterize in vivo delivery of cisplatin immunoconjugates targeting EC caveolae in tumors, and Aim 2 - to assess therapeutic efficacy of the EC caveolae-targeting cisplatin immunoconjugates. This project will utilize our new intravital microscopy (IVM) tumor model system in addition to spontaneous mammary tumors from genetically engineered mice. IVM permits direct visualization of targeting and endothelial processing as well as stroma and tumor cell responses, all of which can be quantified to help provide new insights into therapeutic mechanisms in tumors. Targeting caveolae opens a specific gateway across the restrictive vascular endothelial barrier. It can provide means for enhanced delivery much closer to ideal targeting in order to achieve more effective therapies. We have demonstrated that AnnA1 is expressed in the vasculature and caveolae of human primary and metastatic lung tumors. Overall, our proposed project could create a paradigm shift away from the passive transvascular delivery that greatly limits drug efficacy in humans, affording the first prototype of caveolae-targeted anticancer therapeutics for future translation into the clinic.