Exosome-mediated mechanisms of metastatic disease in non-small cell lung cancer

肺癌是美国退伍军人癌症死亡的主要原因，转移性疾病是压倒性的。 死亡的主要原因。基于我们的初步发现，我们假设来源于 高度活跃的癌前细胞可以通过旁分泌将转移表型转移到邻近的正常细胞 发信号。在当前的退伍军人事务部（VA）Merit Review资助期间，我们确定并选择了一个 观察到高度迁移的癌前气道上皮细胞亚群通过 微尺度收缩的速率高达对照细胞的100倍。这些高度迁移的细胞 显示增强的体内转移行为。我们从这些高密度的细胞中分离并鉴定了外泌体， 迁移性癌前人类气道上皮细胞，以及恶性胸腔积液（MPE）衍生的 转移性癌细胞在癌前病变和转移性疾病的两种情况下，含有独特的外泌体 组学标记将转移表型转移到非运动细胞。从这些签名，我们有 鉴定了介导转移行为和免疫抑制的转移的潜在候选物。到 为了促进转移性疾病的研究，我们将利用我们的MPE生物库进行外泌体分离， 图2示出了来自个体患者的MPE衍生的转移性肺癌细胞的表征。虽然已知外泌体 作为细胞相互作用的功能介质，导致癌症转移，这将是第一个 一项全面研究，以充分描述这些事件在肺部疾病背景下的机制。 癌前迁移，以及在从MPE转移细胞的情况下。为了表征和 了解这一进程在调节促进移徙能力方面的机制， 在癌前病变和转移性肺癌细胞中的免疫抑制，我们将：1）确定 外泌体，通过表征外泌体在NSCLC疾病谱中的作用， 来源于癌前病变和转移癌细胞的外泌体货物，2）确定 外泌体依赖性促进上皮间质转化（EMT）和上皮细胞迁移能力 和肺癌细胞，以及3）确定外泌体依赖性免疫抑制的机制。而 据报道，外来体在导致癌症的细胞相互作用中充当功能性介质 转移，这将是第一个全面的研究，以充分表征机制的基础上充分 在肺癌前转移和转移性肺癌的背景下， 本研究的外泌体介导的转移调节的背景下，癌前病变和转移是独特的 并将有助于定义隐匿性转移的生物学，提供对平行转移现象的见解。 进展，增加生物标志物发现的可能性，并为肺癌拦截提供靶点。

Lung cancer is the leading cause of cancer death among US Veterans, and metastatic disease is overwhelmingly the predominant cause of death. Based on our preliminary findings, we hypothesize that exosomes derived from highly motile premalignant cells can transfer metastatic phenotypes to neighboring normal cells via paracrine signaling. In the current Veteran Affairs (VA) Merit Review funding period, we identified and selected a subpopulation of highly migratory premalignant airway epithelial cells that were observed to migrate through microscale constrictions at up to 100-fold the rate of the control unselected cells. These highly migratory cells demonstrate enhanced metastatic behavior in vivo. We isolated and characterized exosomes from these highly migratory premalignant human airway epithelial cells, as well as, malignant pleural effusion (MPE)-derived metastatic cancer cells. In both settings of premalignancy and metastatic disease, exosomes containing unique omic signatures transferred the metastatic phenotype to non-motile cells. From these signatures, we have identified potential candidates mediating the transfer of metastatic behavior and immunosuppression. To facilitate the study of metastatic disease, we will utilize our MPE-biobank for exosome isolation and characterization from individual patient's MPE-derived metastatic lung cancer cells. While exosomes are known to serve as functional mediators in cell interaction leading to cancer metastasis, this will be the first comprehensive study to fully characterize the mechanisms underlying these events in the context of pulmonary premalignant migration, as well as, in the context of metastatic cells from MPEs. In order to characterize and understand the mechanisms of this process in regulating the promotion of migratory capacity and immunosuppression in premalignant and metastatic lung cancer cells we will: 1) Determine the capacity of exosomes to drive disease progression across the spectrum of disease in NSCLC by characterizing the exosomal cargo derived from premalignant and metastatic cancer cells, 2) Determine the mechanisms of exosome-dependent promotion of epithelial mesenchymal transition (EMT) and migratory capacity of epithelial and lung cancer cells and, 3) determine the mechanisms of exosome-dependent immune suppression. While exosomes have been reported to serve as functional mediators in cellular interaction leading to cancer metastasis, this will be the first comprehensive study to fully characterize the mechanisms underlying the full spectrum of disease, both in the context of pulmonary premalignant migration as well as metastatic lung cancer. This study of exosome mediated metastatic-modulation in the context of premalignancy and metastasis is unique and will help define the biology of occult metastasis, provide insights into the phenomenon of parallel progression, increase the possibility of biomarker discovery and provide targets for lung cancer interception.