Metabolic tagging of tumor exosomes for developing enhanced exosome vaccines

肿瘤外泌体的代谢标记用于开发增强型外泌体疫苗

• 批准号: 10645558
• 负责人: Hua Wang
• 金额: $ 16.89万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10645558
• 关键词: Adjuvant; Agonist; Antigen Presentation; Antigen-Presenting Cells; Antigens; Azides; Binding; Cancer Vaccines; Cell membrane; Chemicals; Chemistry; Clinical; Clinical Trials; Cytotoxic T-Lymphocytes; Dendritic Cells; Dendritic cell activation; Development; Endosomes; Generations; Glioblastoma; Glycoengineering; Glycolipids; Glycoproteins; Goals; Immunomodulators; Immunotherapy; Inherited; Label; Ligands; Mediating; Membrane; Membrane Glycoproteins; Metabolic; Modeling; Mus; Polysaccharides; Process; Proteins; Safety; Source; Surface; TLR3 gene; TLR7 gene; TLR8 gene; Technology; Testing; Therapeutic; Toll-like receptors; Tumor Antigens; Tumor-Derived; Vaccination; Vaccines; cancer cell; cancer therapy; cancer type; clinical translation; efficacy evaluation; exosome; extracellular vesicles; immune modulating agents; improved; malignant breast neoplasm; nanosized; neoplastic cell; pre-clinical; preclinical study; response; success; sugar; triple-negative invasive breast carcinoma; tumor; uptake

SUMMARY Immunotherapies have shifted the paradigm for cancer treatment in the past decade. Among them, therapeutic cancer vaccines, consisting of tumor antigens and adjuvants for activating antigen presenting cells (e.g., dendritic cells (DCs)), have shown the promise to elicit persistent humoral and cytotoxic T lymphocyte (CTL) response, but are limited by the modest therapeutic benefit. Hurdles for developing potent cancer vaccines include the lack of available tumor antigens and sub-optimal modulation of DCs. Tumor-derived exosomes, the nano-sized extracellular vesicles secreted by tumor cells, are a good source of tumor antigens and have been widely explored as cancer vaccines in preclinical studies and clinical trials. However, the antitumor efficacy is far from satisfactory, likely as a result of poor CTL response. The incorporation of adjuvants that can bind to the toll like receptors on DC membrane and thus activate DCs has become standard practice for improving conventional vaccines, but simple mixing with adjuvants has failed to enhance the antitumor efficacy of tumor exosome vaccines. The primary goal of this project is to develop a strategy that can well integrate tumor exosomes and adjuvants for the development of potent tumor exosome vaccines. To achieve this, we utilize a metabolic glycan labeling approach to label glycoproteins and glycolipids on the membrane of cancer cells with chemical tags (e.g., azido groups), and hypothesize that exosomes secreted by these labeled cancer cells will carry chemical tags on the surface. The chemically tagged tumor exosomes can covalently capture adjuvants and other immunomodulatory agents via efficient and bioorthogonal click chemistries, potentially resulting in improved activation and antigen presentation of DCs and enhanced CTL response. This project will be organized around two specific aims. In Aim 1, metabolic glycan labeling of various types of cancer cells for the generation of azido- labeled exosomes will be explored. Two hypotheses will be tested in this aim: (1) azido-labeled tumor cells can secrete azido-labeled exosomes; (2) the azido groups on the surface of exosomes can efficiently conjugate dibenzocyclooctyne (DBCO)-molecules via click chemistry. In Aim 2, adjuvant-conjugated tumor exosomes will be explored as therapeutic cancer vaccines, with the resulting CTL response and antitumor efficacy evaluated in murine breast cancer and glioblastoma models. Tumor exosomes conjugated with both adjuvants and DC- targeting ligands (e.g., anti-DEC205) will also be developed and evaluated. Successful completion of the proposed studies not only will provide a generalizable technology to generate chemically tagged exosomes from tumor cells for tracking and targeting purposes, but will also result in the development of potent tumor exosome vaccines with great potential for clinical translation.

概括 在过去的十年中，免疫疗法改变了癌症治疗的范式。其中，治疗性 癌症疫苗，由肿瘤抗原和用于激活抗原呈递细胞（例如树突状细胞）的佐剂组成 细胞（DC）），已显示出引发持续体液和细胞毒性 T 淋巴细胞（CTL）反应的希望， 但受到有限的治疗效果的限制。开发有效的癌症疫苗的障碍包括缺乏 可用的肿瘤抗原和 DC 的次优调节。肿瘤来源的外泌体，纳米尺寸 肿瘤细胞分泌的细胞外囊泡是肿瘤抗原的良好来源，已被广泛研究 在临床前研究和临床试验中探索作为癌症疫苗。但其抗肿瘤功效却远未达到 令人满意，可能是由于 CTL 反应不佳所致。加入可以与收费物结合的佐剂，例如 DC膜上的受体从而激活DC已成为改善传统方法的标准做法 疫苗，但简单与佐剂混合并不能增强肿瘤外泌体的抗肿瘤功效 疫苗。该项目的首要目标是开发一种能够很好地整合肿瘤外泌体和 用于开发强效肿瘤外泌体疫苗的佐剂。为了实现这一目标，我们利用代谢聚糖 用化学标签在癌​​细胞膜上标记糖蛋白和糖脂的标记方法 （例如叠氮基），并假设这些标记的癌细胞分泌的外泌体将携带化学物质 表面上的标签。化学标记的肿瘤外泌体可以共价捕获佐剂和其他 通过有效和生物正交点击化学的免疫调节剂，可能会导致改善 DC 的激活和抗原呈递以及增强的 CTL 反应。该项目将围绕 两个具体目标。在目标 1 中，对各种类型的癌细胞进行代谢聚糖标记，以产生叠氮基- 将探索标记的外泌体。在此目的中将测试两个假设：（1）叠氮基标记的肿瘤细胞可以 分泌叠氮基标记的外泌体； (2)外泌体表面的叠氮基可以有效缀合 通过点击化学生成二苯并环辛炔 (DBCO) 分子。在目标 2 中，佐剂缀合的肿瘤外泌体将 被探索作为治疗性癌症疫苗，并评估由此产生的 CTL 反应和抗肿瘤功效 在小鼠乳腺癌和胶质母细胞瘤模型中。与佐剂和 DC-缀合的肿瘤外泌体 靶向配体（例如抗DEC205）也将被开发和评估。圆满完成了 拟议的研究不仅将提供一种通用技术来生成化学标记的外泌体 用于跟踪和靶向肿瘤细胞，但也将导致有效的肿瘤外泌体的发展 具有巨大临床转化潜力的疫苗。