A physical sciences approach to investigate the role of exosomes in metastatic progression

研究外泌体在转移进展中的作用的物理科学方法

• 批准号: 10533581
• 负责人: WEI GUO
• 金额: $ 8.24万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-16 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10533581
• 关键词: Address; Affect; Biological Markers; Biomedical Engineering; Breast Cancer Cell; Breast Cancer Model; Breast Cancer Patient; Cell Culture Techniques; Cessation of life; Clinical; Development; Disease; Disseminated Malignant Neoplasm; Extracellular Matrix; Fostering; Foundations; Future; Genetically Engineered Mouse; Immune checkpoint inhibitor; Immune system; Immunity; Immunologic Surveillance; Immunotherapy; In Vitro; Isogenic transplantation; Light; Link; Malignant Neoplasms; Methods; Modeling; Molecular; Neoplasm Metastasis; Non-Invasive Cancer Detection; PD-1/PD-L1; Pathway interactions; Patients; Play; Primary Neoplasm; Production; Role; Signal Transduction; Site; T-Lymphocyte; Testing; Therapeutic Intervention; Time; Tissues; Tumor Immunity; Tumor Tissue; Tumor-Derived; United States; Woman; anticancer research; base; biomarker development; biophysical model; cancer cell; clinically relevant; exosome; experimental study; immunoregulation; in vivo; interest; liquid biopsy; macrophage; metastasis prevention; mortality; neoplastic cell; pharmacokinetic model; physical science; trafficking; translational potential; triple-negative invasive breast carcinoma; tumor; tumor immunology; tumor progression

Metastatic cancer is a major clinical challenge that accounts for numerous deaths annually in the United States, particularly in women with triple-negative breast cancer (TNBC). Many tumors develop within a microenvironment (TME) characterized by altered/stiffened extracellular matrix (ECM) and compromised immunity. These alterations play a causal role in malignancy and metastasis. Recently tumor-derived exosomes have drawn tremendous interest as they are implicated in modulating the TME, suppressing anti-tumor immunity, and preparing the metastatic site for progression. A hallmark of cancer cells is their ability to evade the immune system. Exosomes play a pivotal role in the suppression of anti-tumor immunity. In this project, focusing on TNBC, we explore how ECM stiffness and cytoskeletal tension (collectively referred to as tissue tension) regulate exosome production and cargo composition, and how these exosomes contribute to the suppression of anti- tumor immunity and promote metastasis. We pursue a unique set of hypotheses linking tissue tension to exosome production and defining the role of tumor-derived exosomes in immune surveillance and metastatic progression. To test our hypotheses, we have assembled a strong team from UPENN and UCSF, integrating expertise in bioengineering, cancer mechanobiology, and cancer immunology. In Aim 1, we address whether and how the tissue tension affects exosome production and alters exosome cargo in vitro in TNBC cells. We will also delineate a molecular pathway linking ECM stiffness to intracellular signaling and exosome trafficking, using experimental and subcellular biophysical modeling methods. In Aim 2, we address how tissue tension promotes metastatic progression via exosomes in vivo. In this aim we test the hypothesis that the tension of the primary tumor tissue enhances exosome production and alters exosome cargo to promote the dissemination of primary tumor cells and foster their survival and outgrowth at the metastatic site. We will use unique genetically engineered mouse models (GEMMs) and syngeneic TNBC models, and TNBC patient PDXs, combined with multiscale pharmacokinetic modeling. In Aim 3, we address how tissue tension contributes to the suppression of anti-tumor immunity. In this aim, we will investigate the role of exosomes derived from tumors with high tension in stiff ECM TMEs in suppressing anti-tumor immunity through (1) reprogramming macrophages against T cells; and (2) the engagement of PD-1/PD-L1 checkpoint axis in T cells. We will use a combination of in vitro cell culture experiments, in vivo genetically engineered mouse models and syngeneic transplant manipulations and tissue-scale agent-based modeling. The expected results will shed light on the roles of exosomes in immune regulation and metastatic tumor progression; these are important and timely questions in cancer research. The results will lay the foundation for future therapeutic intervention of metastatic disease through the identification of actionable biomarkers, development of new immune checkpoint inhibitor (ICB)-based therapies, and ultimately reduce patient mortality.

转移性癌症是一个主要的临床挑战，每年在美国造成许多死亡。 特别是在患有三阴性乳腺癌（TNBC）的女性中。许多肿瘤发生在 微环境（TME）的特征是细胞外基质（ECM）改变/硬化， 免疫力这些改变在恶性肿瘤和转移中起因果作用。最近肿瘤来源的外泌体 已经引起了极大的兴趣，因为它们涉及调节TME，抑制抗肿瘤免疫， 并使转移部位准备好进行。癌细胞的一个标志是它们逃避免疫系统的能力。 系统外泌体在抑制抗肿瘤免疫中起关键作用。在这个项目中，重点是 TNBC，我们探讨ECM刚度和细胞骨架张力（统称为组织张力）如何调节 外泌体产生和货物组成，以及这些外泌体如何有助于抑制抗- 肿瘤免疫和促进转移。我们追求一套独特的假设，将组织张力与 外泌体产生和确定肿瘤来源的外泌体在免疫监视和转移中的作用 进展为了验证我们的假设，我们组建了一支来自UPENN和UCSF的强大团队， 生物工程、癌症机械生物学和癌症免疫学方面的专业知识。在目标1中，我们讨论 以及在TNBC细胞中，组织张力如何影响外泌体产生并在体外改变外泌体货物。我们将 还描绘了一个分子途径，将ECM硬度与细胞内信号传导和外泌体运输联系起来，使用 实验和亚细胞生物物理建模方法。在目标2中，我们讨论了组织张力如何促进 通过体内外泌体的转移进展。在这个目标中，我们测试的假设，即紧张的主要 肿瘤组织增强外泌体产生并改变外泌体货物以促进原发性肿瘤细胞的传播， 肿瘤细胞并促进它们在转移部位的存活和生长。我们将使用独特的基因 工程化小鼠模型（GEMM）和同基因TNBC模型，以及TNBC患者PDX，与 多尺度药代动力学建模在目标3中，我们讨论了组织张力如何有助于抑制 抗肿瘤免疫在这个目标中，我们将研究来源于肿瘤的外泌体在高张力下的作用， 在刚性ECM TME中通过（1）针对T细胞重编程巨噬细胞来抑制抗肿瘤免疫; 以及（2）T细胞中PD-1/PD-L1检查点轴的接合。我们将使用体外细胞 培养实验、体内基因工程小鼠模型和同系移植操作， 组织尺度的基于Agent的建模。预期的结果将阐明外泌体在免疫系统中的作用。 调节和转移性肿瘤进展;这些是癌症研究中重要而及时的问题。的 结果将为将来通过鉴定转移性疾病的治疗干预奠定基础。 可行的生物标志物，开发新的免疫检查点抑制剂（ICB）为基础的疗法，并最终 降低患者死亡率。