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.

总结 免疫疗法在过去十年中改变了癌症治疗的范式。其中，治疗 由肿瘤抗原和用于活化抗原呈递细胞的佐剂组成的癌症疫苗（例如，树突状 细胞（DC）），已经显示出引起持续的体液和细胞毒性T淋巴细胞（CTL）应答的前景， 但受到适度治疗益处的限制。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，纳米大小的 由肿瘤细胞分泌的细胞外囊泡是肿瘤抗原的良好来源， 在临床前研究和临床试验中探索作为癌症疫苗。然而，其抗肿瘤功效远未达到 这可能是由于CTL应答较差所致。掺入可以结合Toll样的佐剂， DC膜上的受体并因此激活DC已经成为改善常规DC的标准实践。 疫苗，但与佐剂简单混合未能增强肿瘤外泌体的抗肿瘤功效 疫苗。该项目的主要目标是开发一种能够很好地整合肿瘤外泌体和 用于开发有效的肿瘤外泌体疫苗的佐剂。为了实现这一点，我们利用代谢聚糖 用化学标签标记癌细胞膜上的糖蛋白和糖脂的标记方法 (e.g.,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， 表面的标签。化学标记的肿瘤外泌体可以共价捕获佐剂和其他佐剂。 通过有效的和生物正交的点击化学，免疫调节剂，潜在地导致改善的 DC的活化和抗原呈递以及增强的CTL应答。该项目将围绕 两个具体目标。在目的1中，各种类型的癌细胞的代谢聚糖标记用于叠氮基- 将探索标记的外泌体。为此，将检验两个假设：（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地缀合 二苯并环辛炔（DBCO）-分子。在目标2中，缀合有抗癌剂的肿瘤外泌体将 作为治疗性癌症疫苗进行探索，并评估由此产生的CTL应答和抗肿瘤功效 在小鼠乳腺癌和胶质母细胞瘤模型中。肿瘤外泌体与佐剂和DC-100两者缀合。 靶向配体（例如，抗DEC 205）也将被开发和评估。成功完成 拟议的研究不仅将提供一种可推广的技术， 肿瘤细胞进行跟踪和靶向的目的，但也将导致发展强大的肿瘤外泌体 具有巨大临床转化潜力的疫苗。