REGULATION OF TRANSCRIPTIONAL COMPETENCE BY CHROMIUM

六价铬(Cr(VI))是ATSDR/EPA优先考虑的前20种化合物之一，因为它促进间质肺纤维化，诱发哮喘，并被公认为可能的人类肺癌致癌物。尽管有流行病学证据表明吸入铬(VI)后会引起职业性和环境性肺部疾病，但很少有研究明确铬(VI)暴露肺的病理变化的细胞和分子基础。总体目标：本研究的总体目标是明确非细胞毒性浓度的铬(VI)改变呼吸道和肺泡上皮细胞诱导性细胞因子和促纤维化基因表达的分子信号机制。我们最近定义了一种新的途径，通过促进必要的共激活因子CREB结合蛋白(CBP)向c-Jun.募集，从而抑制转录因子NF-kappaB的转录能力。我们假设，铬(VI)诱导的共激活因子募集的改变改变了可诱导基因的表达谱，并增强了Fas诱导的细胞凋亡。铬(VI)的这些作用有利于肺纤维化的发生。具体目标：赠款的目标1将使用培养的人肺细胞来定义细胞信号通路，以调节铬(VI)诱导的CBP向特定转录因子(如c-Jun)的募集。Aim 2将使用这些模型来证明CBP的竞争是铬(VI)诱导的核因子-kappaB反式激活丧失的限速步骤。CBP依赖的转录因子能力的改变对Fas诱导的细胞凋亡和基因表达的影响将被检测。最终目的将使用正常和独特的转基因小鼠模型来检测体内铬(VI)诱导的转录因子活性的调节以及可诱导的促凋亡和促纤维化的表型变化。意义：这些研究将确定可诱导基因表达改变和暴露于铬(VI)后增加呼吸道和肺泡上皮细胞对凋亡的易感性的基本表观遗传学机制。现在认为，呼吸道细胞凋亡是肺纤维化的一种潜在的非炎症机制。因此，这些研究将把细胞培养中的观察转化为体内暴露模型，以极大地提高对铬(VI)如何促进肺部疾病的基本理解。

Hexavalent chromium (Cr(VI)) is in the top 20 compounds of the ATSDR/EPA priority list since it promotes interstitial lung fibrosis, induces asthma, and is recognized as a probable human lung carcinogen. Despite the epidemiological evidence for both occupationally and environmentally-derived pulmonary disease following inhalation of Cr(VI), there are few studies that define the cellular and molecular basis for pathologic changes in the Cr(VI)-exposed lung. Overall Objective: The overall objective of the proposed studies is to define the molecular signaling mechanisms through which non-cytotoxic concentrations of Cr(VI) alter inducible cytokine and profibrotic gene expression in airway and alveolar epithelial cells. We have recently defined a novel pathway through which Cr(VI) inhibits the transcriptional competence of the transcription factor NF-kappaB by promoting recruitment of the essential co-activator CREB-binding protein (CBP) to c-Jun. We hypothesize that Cr(VI)-induced alteration of co-activator recruitment changes the profile of inducible gene expression and potentiates Fas-induced apoptosis. These effects of Cr(VI) favor development of pulmonary fibrosis. Specific Objectives: Aim 1 of the grant will use human lung cells in culture to define cellular signaling pathways that regulate Cr(VI)-induced recruitment of CBP to specific transcription factors (e.g. c-Jun). Aim 2 will use these models to demonstrate that competition for CBP is the rate limiting step in Cr(VI)- induced loss of NF-kappaB transactivation. The consequence of a switch in CBP-dependent transcription factor competence on FAS- induced apoptosis and gene expression will be examined. The final Aim will use normal and unique transgenic mouse models to examine Cr(VI)-induced regulation of transcription factor activity and of inducible pro-apoptotic and pro-fibrotic phenotypic changes in vivo. Significance: The studies will define fundamental epigenetic mechanisms for altered inducible gene expression and increased susceptibility of airway and alveolar epithelial cells to apoptosis following exposure to Cr(VI). Apoptosis of the airway is now recognized as an underlying non-inflammatory mechanism for lung fibrosis. Thus, these studies will translate observations in cell culture into in vivo models of exposure to greatly improve the basic understanding of how Cr(VI) promotes pulmonary diseases.