Characterization of cinnabarinic acid mediated cytoprotection upon liver injury

 描述（由申请方提供）：过度或持续的肝细胞凋亡是各种急性肝损伤（如暴发性肝炎、缺血和再灌注损伤、药物诱导的肝毒性和与病毒性肝炎、非酒精性脂肪肝和酒精性肝病相关的慢性损伤）的潜在病理学。该提案建立在 我最近观察到，芳烃受体（AhR）激动剂朱砂酸（CA）上调AhR靶基因斯钙素2（STC 2）的表达，该基因对ER/氧化应激诱导的肝细胞凋亡起关键的细胞保护作用。此外，STC 2诱导发生在响应内源性AhR激动剂朱砂酸，但不是典型的外源性AhR激动剂2，3，7，8-四氯二苯并-p-二恶英（TCDD）。本申请的具体目的1测试内源性和外源性AhR激动剂可以诱导具有不同生理后果的二分信号传导级联的假设。我将专门研究CA，而不是TCDD，诱导STC 2表达负责细胞保护的机制。具体目标2开始内源性AhR激动剂CA的全基因组功能评估。这项研究将特别确定使用下一代测序CA调节AhR靶基因的库。具体目标3将在慢性酒精诱导的肝毒性小鼠模型中测试CA的体内细胞保护特性。这一目标将解决潜在的CA作为一个先导化合物在治疗上有用的药物对肝损伤的发展。该研究项目与结构化的职业发展计划相结合。我的职业发展计划的目标是：1）发展一个独立的研究计划，在AhR生物学为中心，了解受体的作用，在肝损伤和激动剂特异性AhR信号。2)通过获得质谱分析、下一代测序分析、肝功能测试和补充生化技术方面的高级培训，扩展我的技能。3)与肝病、细胞凋亡、药物开发、生物物理学和生物信息学领域的研究人员建立合作关系。4)确保高质量的出版物，演示文稿的跟踪记录，并成为竞争对手发起的资金，以开发AhR生物学计划。这项研究意义重大，因为它将决定 STC 2的激动剂特异性表达和功能的分子基础，鉴定CA诱导的具有生理学重要作用的AhR依赖性靶基因，并将作为开发CA作为未来细胞保护治疗剂的先导化合物的基础。为了实现我的研究和职业发展目标，我将接受由校外资助的高级研究人员和技术专家组成的跨学科指导团队的指导。这种指导与其他丰富的活动，包括研讨会，课程，赠款写作和技术讲习班相结合。在获奖期间获得的指导和培训的结合，再加上一个令人兴奋的研究计划的成熟，即-CA介导的AhR依赖性细胞保护反应肝损伤的表征-将赋予我必要的专业知识，以建立一个独立的研究者的职业生涯。

 DESCRIPTION (provided by applicant): Excessive or sustained hepatocyte apoptosis is an underlying pathology of various acute liver injuries like fulminant hepatitis, ischemia and reperfusion damage, drug induced hepatotoxicity and chronic injuries associated with viral hepatitis, non-alcoholic fatty liver disease and alcoholic liver disease. This proposal is built on my recent observation that the aryl hydrocarbon receptor (AhR) agonist cinnabarinic acid (CA) upregulates expression of AhR target gene, Stanniocalcin 2 (STC2), which plays a crucial cytoprotective role against ER/oxidative stress induced apoptosis in hepatocytes. Moreover, STC2 induction occurs in response to the endogenous AhR agonist cinnabarinic acid, but not the prototypical exogenous AhR agonist 2,3,7,8- tetrachlorodibenzo-p-dioxin (TCDD). Specific aim 1 of this application tests the hypothesis that endogenous and exogenous AhR agonists can induce dichotomous signaling cascades with distinct physiological consequences. I will specifically examine mechanism by which CA, but not TCDD, induces STC2 expression responsible for cytoprotection. Specific aim 2 embarks on genome-wide functional assessment of endogenous AhR agonist CA. This study will particularly identify repertoire of CA regulated AhR target genes using next generation sequencing. Specific Aim 3 will test the cytoprotective property of CA in vivo in a mouse model of chronic alcohol induced hepatotoxicity. This aim will address potential of CA as a lead compound in the development of therapeutically useful drugs against liver injury. This research project is integrated with a structured career development plan. The objectives of my career development plan are: 1) To develop an independent research program in AhR biology centered on understanding receptor's role in liver injury and agonist specific AhR signaling. 2) Expand my skill sets by obtaining advanced training in mass spectrometry, next generation sequencing analysis, liver function tests and complementary biochemical techniques. 3) Establish collaborative relationships with researchers in liver diseases, apoptosis, drug development, biophysics and bioinformatics fields. 4) Secure a track record of high quality publications, presentations and become competitive for investigator-initiated funding to develop program in AhR biology. This study is significant as it will determine molecular basis for the agonist specific expression and function of STC2, identify CA induced AhR dependent target genes with physiologically important roles, and will serve as a foundation for developing CA as a lead compound in the development of future cytoprotective therapeutics. To accomplish my research and career development objectives, I will receive guidance from an interdisciplinary mentoring team of extramurally funded senior investigators and technical experts. This mentorship is integrated with other enrichment events including seminars, courses, grant writing and technical workshops. The combination of mentoring and training obtained during the award period coupled with the maturation of an exciting research program namely - characterization of CA mediated AhR dependent cytoprotection in response to liver injury - will endow me with the requisite expertise to establish a career as an independent investigator.