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首要原因，转移性疾病占压倒性多数 最主要的死因。根据我们的初步发现，我们假设外显体起源于 高度活动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。在当前退伍军人事务部(VA)功绩审查资助期，我们确定并选择了 观察到的高度迁移的癌前呼吸道上皮细胞亚群 微尺度收缩的速率高达对照未选择细胞的100倍。这些高度迁移的细胞 在体内表现出增强的转移行为。我们从这些高度敏感的生物中分离并鉴定了外切体 移行性癌前病变上皮细胞以及恶性胸腔积液(MPE)来源 转移性癌细胞。在癌前疾病和转移性疾病的环境中，包含独特的 基因组特征将转移表型转移到非运动性细胞。从这些签名中，我们有 确定了调节转移行为和免疫抑制转移的潜在候选者。至 为了方便对转移疾病的研究，我们将利用我们的MPE-生物库来分离和 来自个体患者MPE来源的转移性肺癌细胞的特征。虽然外显体是已知的 为了在导致癌症转移的细胞相互作用中充当功能介体，这将是第一次 全面研究，以充分描述这些事件背后的机制在肺 癌前迁移，以及在MPE转移细胞的背景下。为了刻画和描述 了解这一过程在调节促进迁徙能力和 癌前病变和转移性肺癌细胞的免疫抑制我们将：1)确定 外切体通过表征非小细胞肺癌疾病谱来推动疾病进展 来自癌前和转移癌细胞的胞外物，2)决定了肿瘤的机制 外切体依赖促进上皮细胞间充质转化和迁移能力的研究 和肺癌细胞，以及3)确定外切体依赖的免疫抑制的机制。而当 据报道，外切体在导致癌症的细胞相互作用中起着功能中介作用。 转移，这将是第一个全面的研究，以充分表征潜在的机制充分 在肺癌前转移和转移性肺癌的背景下，疾病的范围都是如此。 在癌前病变和转移的背景下，外切体介导的转移调控的研究是独一无二的 将有助于定义隐匿性转移的生物学，为平行转移现象提供洞察力 进展，增加发现生物标记物的可能性，并为肺癌的阻断提供靶点。

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.