Stat3 Downstream Genes as Lung Adenocarcinoma Biomarkers

描述（由申请人提供）：这项工作的长期目标是确定用于动物和人类肺腺癌诊断和预后的分泌蛋白生物标志物。除了气体交换，肺也是宿主通过炎症反应进行防御的器官。炎症是促进病原体清除和修复肺组织损伤的保护过程。然而，过度的炎症会导致严重的后果，并导致致癌。在肺部炎症中，一个常见的促炎基因组是白细胞介素6 （IL-6）家族细胞因子。在与细胞表面受体结合后，IL-6家族成员通过Janus-activated kinase （JAKs）触发信号换能器和转录激活因子3 （Stat3）的激活（磷酸化）。为了评估STAT3在肺中持续激活的后果，我们实验室最近建立了一个多西环素控制的CCSP-rtTA/(tetO)7-CMV-Stat3C双基因小鼠模型，该模型在肺泡II型（AT II）上皮细胞中过表达STAT3C （STAT3的构成活性形式）。在连续的步骤中，Stat3C过表达以高频率（80%）促进肺部炎症和腺癌。这支持了Stat3信号触发的持续炎症诱导肺腺癌的概念。在寻找人类肺癌样本中，Stat3上调与腺癌和鳞状细胞癌高度相关。因此，Stat3及其下游基因可作为动物和人类肺癌诊断的生物标志物。通过Affymetrix基因芯片分析，在CCSP-rtTA/(tetO)7-CMV-Stat3C双转基因小鼠中鉴定出多个Stat3下游基因。从这个列表中，一组变化最多的基因在人类腺癌中也出现了类似的变化，证实Stat3下游基因适合肺癌的诊断和预后。由于许多Stat3下游基因是血浆蛋白，我们计划将它们用于动物肺腺癌模型和人类肺癌患者的血液检测。该提案的中心假设是Stat3下游基因的分泌蛋白产物可用于动物模型和人类肺腺癌的诊断和预后。为了验证中心假设，我们提出了两个目标：1)在动物中鉴定Stat3下游基因作为肺癌生物标志物。将使用三种与炎症相关的肺腺癌动物模型。虽然它们代表不同的诱导肺腺癌的分子机制，但Stat3基因的上调和激活是这三种动物模型的共同机制；2) Stat3下游基因作为人类肺癌生物标志物的研究。将在来自人类腺癌、鳞状细胞癌和小细胞肺癌的肺组织和血液样本中筛选推定的分泌生物标志物。在这些研究之后，我们希望在动物模型和人类肺癌患者的血液中形成一组用于诊断和预后的蛋白质生物标志物。

DESCRIPTION (provided by applicant): The long-term goal of this work is to identify secretory protein biomarkers for lung adenocarcinoma diagnosis and prognosis in animals and in humans. In addition to gas exchange, the lung is an organ for host defense through inflammatory responses. Inflammation is a protective process that facilitates pathogen clearance and repairs tissue injury in the lung. However, exuberant inflammation can cause severe consequences and lead to carcinogenesis. One commonly induced pro-inflammatory gene group during pulmonary inflammation is the interleukin 6 (IL-6) family cytokines. Upon binding to their cell-surface receptors, IL-6 family members trigger activation (phosphorylation) of signal transducer and activator of transcription 3 (Stat3) by Janus-activated kinases (JAKs). To assess the consequences of STAT3 persistent activation in the lung, a doxycycline-controlled CCSP-rtTA/(tetO)7-CMV-Stat3C bitransgenic mouse model was generated recently in our laboratory that over-expresses STAT3C (the constitutively active form of STAT3) in alveolar type II (AT II) epithelial cells. In sequential steps, Stat3C over-expression promoted inflammation and adenocarcinoma in the lung with a high frequency (80%). This supports a concept that persistent inflammation triggered by the Stat3 signaling induces lung adenocarcinoma. In searching for human lung cancer samples, Stat3 up-regulation was highly associated with adenocarcinoma and squamous cell carcinoma. Therefore, Stat3 and its downstream genes can be used as biomarkers for lung cancer diagnosis in animals and humans. By Affymetrix GeneChip microarray analysis, multiple Stat3 downstream genes were identified in CCSP-rtTA/(tetO)7-CMV-Stat3C bitransgenic mice. A set of mostly-changed-genes from this list showed similar changes in human adenocarcinoma, confirming that Stat3 downstream genes are suitable for lung cancer diagnosis and prognosis. Since many Stat3 downstream genes are plasma proteins, we plan to use them for blood testing in both animal lung adenocarcinoma models and in human lung cancer patients. The central hypothesis for this proposal is that secreted protein products of Stat3 downstream genes can be used for diagnosis and prognosis of lung adenocarcinomas in animal models and in humans. Two aims are proposed to test the central hypothesis: 1) Characterization of Stat3 downstream genes as lung cancer biomarkers in animals. Three animal models that developed lung adenocarcinoma in association with inflammation will be used. Although they represent different molecular mechanisms for inducing lung adenocarcinoma, Stat3 gene up-regulation and activation are the common mechanism in all three animal models; 2) Characterization of Stat3 downstream genes as lung cancer biomarkers in humans. Putative secreted biomarkers will be screened in lung tissues and blood samples that are from human adenocarcinoma, squamous cell carcinoma and small cell lung cancer. After these studies, we would like to formulate a panel of protein biomarkers for diagnosis and prognosis in the blood of animal models and in human lung cancer patients. PUBLIC HEALTH RELEVANCE: Lung cancer is one of the biggest public health challenges facing the United States and many other countries. This work will identify protein biomarkers for lung cancer diagnosis and prognosis in humans and in animals.