The Adenosinergic Pathway in Tumor-derived Exosomes

项目摘要/摘要 肿瘤细胞产生和释放大量的细胞外小泡(EV)。小(30-150 nm)的子集 来源于肿瘤细胞内胞腔的EVS被称为Tex(肿瘤衍生的外切体)。特克斯 携带各种免疫调节分子，相对于正常细胞来源的外切体，NTEX(非肿瘤 细胞来源的外切体)，富含抑制蛋白，抑制免疫效应细胞的功能。 TeX最近已经成为一种主要的免疫抑制系统，被肿瘤用来逃离宿主 免疫系统。我们的结果表明，腺苷(ADO)，现在被认为是一个主要的免疫检查点 癌症，驱动腺苷能途径的酶通过以下途径被携带并输送到靶免疫细胞 德州。我们推测TeX介导的对肿瘤免疫细胞的抑制是由于ADO通过 TeX，从而促进肿瘤的生长和转移。我们将首先通过比较 用一种独特的方法从黑色素瘤患者血浆中分离出Tex和NTEX的腺苷能通路活性 我们开发的免疫捕获方法。与新推出的高度灵敏的乙烯桥相配合 化学和最先进的UPLC-MS/MS系统，我们现在有了测量活性的手段 每个患者配对的TeX和NTEX中产生ADO的典型和非典型的生化途径。 我们将把“腺苷能驱动力”定义为TeX产生免疫抑制ADO的能力，并将其与 对每个患者的免疫细胞进行体外功能分析。HIGH目标1中的发现 在黑色素瘤患者中，Tex相对于NTEX的腺苷能驱动有望证实Tex在 肿瘤免疫逃逸。AIM 1还将提供对每个黑色素瘤的腺苷能驱动的体外测量 在AIMS 2和3研究中使用的患者。在目标2中，“高”与“低”腺苷能驱动力的特克斯将是 给小鼠注射静脉注射，以确定其对肿瘤发生、肿瘤生长、血管生成、免疫功能的体内影响 功能障碍和转移形成。这一目标将TeX的作用从免疫抑制扩展到肿瘤 促进成长活动。各种成熟的小鼠模型将被用于测量腺苷能 TeX在体内的活性。我们还将检查体内对腺苷途径的阻断是否能逆转 由德州大学调解的有害影响。在目标3中，我们将评估腺苷能驱动在TEX作为一种 免疫检查点抑制剂预后和反应的生物标志物。利用从银行分离出来的TeX 在黑色素瘤患者的血浆中，我们将把TEX中的腺苷能驱动与疾病分期和 临床病理终点以及预后和对免疫治疗的反应。通过阐明其作用 TeX在ADO介导的免疫抑制和肿瘤进展中的作用，这项研究引入了新的 有望改善癌症治疗和患者生存的治疗靶点和新的癌症生物标志物。

PROJECT SUMMARY/ABSTRACT Tumor cells produce and release large numbers of extracellular vesicles (EVs). A subset of small (30-150 nm) EVs derived from the endocytic compartment of the tumor cell is called TEX (Tumor-derived Exosomes). TEX carry various immunoregulatory molecules and relative to normal cell-derived exosomes, NTEX (Non-Tumor cell-derived Exosomes), are enriched in inhibitory proteins which suppress functions of immune effector cells. TEX have recently emerged as a major immunosuppressive system used by tumors to escape from the host immune system. Our results indicate that adenosine (ADO), now considered a major immune checkpoint in cancer, and the enzymes driving the adenosinergic pathway are carried and delivered to target immune cells by TEX. We hypothesize that TEX-mediated suppression of immune cells in cancer is due to ADO distributed by TEX, which thereby encourage tumor growth and metastasis. We will test this hypothesis first by comparing the adenosinergic pathway activity in TEX versus NTEX isolated from plasma of patients with melanoma by a unique immune capture method we have developed. In concert with newly introduced highly sensitive etheno-bridge chemistry and a state-of-the-art UPLC-MS/MS system, we now have means to measure the activity of the canonical and non-canonical ADO-producing biochemical pathways in paired TEX versus NTEX of each patient. We will define “adenosinergic drive” as the capacity of TEX to produce immunosuppressive ADO and will link it to in vitro functional assays performed with each patient’s immune cells. The discovery in Aim 1 of high adenosinergic drive in melanoma patients TEX relative to NTEX is expected to confirm the major role of TEX in tumor immune escape. Aim 1 will also provide an in vitro measure of adenosinergic drive in every melanoma patient for use in Aims 2 and 3 studies. In Aim 2, TEX with “high” versus “low” adenosinergic drive will be injected IV into mice to determine their in vivo effects on carcinogenesis, cancer growth, angiogenesis, immune dysfunction and metastasis formation. This Aim extends the role of TEX from immune suppression to tumor growth promoting activities. Various well-established mouse models will be used to measure adenosinergic activity of TEX in vivo. We will also examine whether in vivo blocking of the adenosine pathway reverses detrimental effects mediated by TEX. In Aim 3, we will assess the utility of adenosinergic drive in TEX as a biomarker of prognosis and response to immune checkpoint inhibitors. Utilizing TEX isolated from banked plasma of melanoma patients, we will correlate adenosinergic drive in the TEX with disease stage and clinicopathological endpoints as well as with prognosis and response to immunotherapy. By elucidating the role of TEX in ADO-mediated immune suppression as well as tumor progression in cancer, this study introduces new therapeutic targets and new cancer biomarkers that are expected to improve cancer therapy and patient survival.