Collaborative Research: Design of Multifunctional Doubly-Fusogenic Liposomes to Deliver Therapeutics and Diagnostics

合作研究：设计多功能双融合脂质体以提供治疗和诊断

• 批准号: 1207022
• 负责人: Stavroula Sofou
• 金额: $ 39万
• 依托单位: Rutgers University New Brunswick
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1207022&HistoricalAwards=false
• 关键词: Collaborative; Research; Design; Multifunctional; Doubly

ID: MPS/DMR/BMAT(7623) 1207022 PI: Sofou, Stavroula ORG: Rutgers UniversityID: MPS/DMR/BMAT(7623) 1206943 PI: Hall, Carol ORG: NC State UniversityINTELLECTUAL MERIT: The most promising strategy at present to provide effective control of advanced solid cancer is a combination of therapies. A potential component of this combination is antivascular therapy. The goal of this work is to design iv-administered theranostic liposome nanocarriers that can be programmed to target tumor vasculature while sparing healthy sites and to release a chemotherapeutic agent, deliver a radioactive imaging agent or both. This will be accomplished through a combined experimental and theoretical approach to develop highly selective lipid vesicles composed of a new class of bi-lipid membranes that rapidly and extensively release doxorubicin intracellularly or deliver positron emitters. The high killing efficacy of the liposomes is based on a dual fusion mechanism that is activated only upon cellular internalization. The project's innovation lies in the choice of individual base components and the synergistic way that they work together to optimize delivery of the drug to the proper site. An additional innovation is the use of molecular level computer simulation to explore the consequences of various choices of liposome parameters "in silico" before trying them out in the lab, thus reducing the number of trial-and-error steps that would normally characterize this type of work . The base components are the following: PSMA (Prostate Specific Membrane Antigen), which is present on tumor vasculature but not in normal tissue, is the target. An anti-PSMA antibody is the ligand. The liposomes are comprised of two functionalized lipids: PEGylated lipids tethered to anti-PSMA antibodies and lipids functionalized with a fusion peptide that promotes fusion with the endosomal membrane. The mechanisms of delivery are the following. During circulation in the blood, the exposed anti-PSMA antibodies result in selective neovasculature targeting while uniformly distributed PEGylated lipids on the liposome surface mask the fusion peptides. Upon endocytosis of liposomes by tumor endothelial cells, pH-induced lipid phase-separation, and domain formation on liposome membranes activates two fusion mechanisms: (1) The fusion peptides become unmasked and bind to the endosome membrane, and (2) The liposomal domain boundaries serve as sites to nucleate fusion with the endosomal membrane. The net result is that the liposome releases its cargo directly into the cytoplasm of tumor endothelial cells, as opposed to the endosome, avoiding entrapment in the endosomal pathway and subsequent degradation by the lysosome. There are three aims: (1) Develop an experimentally informed general computational tool to facilitate the design of liposomes and to test hypotheses about the role of the different components in the proposed hierarchical assembly. (2) Engineer liposomes containing anti-PSMA ligands and small fusion peptides, and investigate the conditions in which the corresponding functionalities exhibit optimal behavior. (3) Demonstrate that dual-fusion liposomes loaded with doxorubicin and Y-86 exhibit: (a) selective targeting of tumor endothelium analogues, (b) effective release of chemotherapeutics and killing of targeted cells, and (c) delivery of sufficient amounts of Y-86 for diagnostic applications.BROADER IMPACTS: Since advanced solid cancer has no cure, many patients could benefit from the proposed research that aims to develop diagnostic and treatment protocols that significantly extend the life expectancy and improve patients? quality of life. The research pursued here will be supplemented by a strong educational component that includes training of two female graduate students and several undergraduate students assigned to this project, integration of several topics of this research in a newly launched open-ended senior design project, general outreach and mentoring activities for high school students and their teachers, and mentoring activities for women graduate students and faculty across the nation. In particular, the 6-week outreach summer program, which will be conducted at Rutgers University, aims to encourage underrepresented and minority high school students to follow a career in sciences and engineering. The program includes hands-on research training, a series of lectures given by speakers from academia and industry on contemporary issues related to biomaterials, and visits to neighboring pharmaceutical industries. Educational materials will be developed that highlight this research including a power point presentation introducing the basics of nanotechnology and drug delivery via soft materials for dissemination over the web.

ID：MPS/DMR/BMAT（7623）1207022 PI：Sofou，Stavroula 组织机构：罗格斯大学ID：MPS/DMR/BMAT（7623）1206943 PI：霍尔，卡罗尔 ORG：北卡罗来纳州立大学智力优势：目前有效控制晚期实体癌最有希望的策略是联合治疗。 这种组合的一个潜在组成部分是抗血管治疗。 这项工作的目标是设计静脉给药的治疗诊断脂质体纳米载体，可以编程为靶向肿瘤血管系统，同时保留健康部位，并释放化疗剂，提供放射性成像剂或两者兼而有之。 这将通过结合实验和理论的方法来实现，以开发由一类新的双脂质膜组成的高选择性脂质囊泡，其快速且广泛地在细胞内释放阿霉素或递送正电子发射体。 脂质体的高杀伤效力基于仅在细胞内化时被激活的双重融合机制。 该项目的创新在于选择单个基础成分，以及它们协同工作的协同方式，以优化药物向适当部位的输送。 另一个创新是使用分子水平的计算机模拟来探索脂质体参数的各种选择的后果，然后在实验室中尝试它们，从而减少了通常表征这种类型工作的试错步骤的数量。 基本成分如下：PSMA（前列腺特异性膜抗原）是靶点，它存在于肿瘤血管系统中，但不存在于正常组织中。 抗PSMA抗体是配体。 脂质体由两种功能化脂质组成：与抗PSMA抗体连接的PEG化脂质和用促进与内体膜融合的融合肽功能化的脂质。 交付机制如下。 在血液循环期间，暴露的抗PSM抗体导致选择性的新血管靶向，而脂质体表面上均匀分布的PEG化脂质掩蔽融合肽。 在脂质体被肿瘤内皮细胞内吞后，pH诱导的脂质相分离和脂质体膜上的结构域形成激活了两种融合机制：（1）融合肽变得暴露并结合到内体膜，以及（2）脂质体结构域边界充当与内体膜成核融合的位点。 净结果是脂质体将其货物直接释放到肿瘤内皮细胞的细胞质中，而不是内体中，避免了内体途径中的截留和随后的溶酶体降解。 有三个目标：（1）开发一种实验性的通用计算工具，以促进脂质体的设计，并测试关于不同组分在所提出的分层组装中的作用的假设。 (2)设计含有抗PSMA配体和小融合肽的脂质体，并研究相应功能表现出最佳行为的条件。 (3)证明负载阿霉素和Y-86的双融合脂质体表现出：（a）选择性靶向肿瘤内皮类似物，（B）有效释放化疗药物并杀死靶细胞，以及（c）递送足够量的Y-86用于诊断应用。更广泛的影响：由于晚期实体癌无法治愈，许多患者可以从拟议的研究中受益，该研究旨在开发显着延长预期寿命的诊断和治疗方案改善患者？生活质量 在此进行的研究将得到一个强有力的教育组成部分的补充，其中包括培训两名女研究生和几名本科生分配到这个项目，整合本研究的几个主题在一个新推出的不限成员名额的高级设计项目，一般的推广和辅导活动，高中学生和他们的教师，并辅导活动，为全国各地的女研究生和教师。 特别是，将在罗格斯大学进行的为期6周的暑期外展计划，旨在鼓励代表性不足和少数民族高中生从事科学和工程职业。 该计划包括动手研究培训，由来自学术界和工业界的演讲者就与生物材料相关的当代问题发表的一系列演讲，以及对邻近制药行业的访问。 将开发重点介绍这项研究的教育材料，包括介绍纳米技术基础知识和通过软材料进行药物输送的幻灯片演示，以便在网络上传播。