Excellence in Research: Stress-induced expression and release of DAMPs: Regulatory role of Epigenetic Factors and Post-translational modifications

近年来得到支持的一种观点是，DNA的表观遗传变化和蛋白质的自然细胞修饰可能会影响应激诱导的免疫反应。细胞应激的主要后果之一是危险相关分子模式（DAMPS）的产生和分泌。DAMPs，包括高迁移率组盒蛋白1 （HMGB1）和热休克蛋白70 (HSP70)，通过与模式识别受体蛋白（能够识别DAMP分子的蛋白）相互作用诱导哺乳动物炎症。早期的研究提供了关于应激细胞分泌HMGB1的化学修饰蛋白的可能作用的证据。然而，我们不知道这些修饰是否也会影响HSP70的分泌。在这个项目中，研究人员将使用计算和细胞生物学方法来测量在五氯酚（一种广泛使用的有机氯除草剂和木材防腐剂）的影响下，肺和肝细胞释放DAMPs的DNA和蛋白质的化学变化。结合计算方法来理解这些基本的细胞机制将极大地改善HBCU参与校区，巴吞鲁日的南方大学和路易斯安那州的格兰布林州立大学所采用的研究方法。该项目的一个重要亮点是，通过为这些HBCU学生提供合格和训练有素的HBCU博士生的教师指导培训，增加非裔美国本科生进入研究科学家职业的人数，这也将培养后者的指导能力。这项卓越研究奖将使研究人员能够开发和建立一个严格的、高质量的研究项目，这将显著改善路易斯安那州少数民族学生的研究经验和教育。本研究旨在探讨应激条件下细胞模型中危险相关分子模式（Danger-Associated molecular Patterns, DAMPs）内源性表达和细胞外释放的相关分子机制。DAMPs包括高迁移率组盒蛋白1 （HMGB1）和热休克蛋白70 （HSP70）。这些蛋白通过与细胞表面或细胞质模式识别受体的相互作用诱导炎症。早期的研究提供了翻译后修饰（PTM）在应激细胞分泌HMGB1中的可能作用的证据，但对HSP70的分泌机制知之甚少。该项目的研究人员将使用计算和细胞生物学方法来确定表观遗传因素和ptm，这些因素和ptm负责五氯酚（一种有机氯除草剂和木材防腐剂）对肺和肝细胞释放DAMPs的影响。这将部分通过测量编码DAMPs基因启动子区域CpG序列上DNA的表观遗传甲基化，以及测量相同启动子区域的组蛋白修饰来完成。进一步结合计算方法来理解基本的细胞机制，将改变HBCU两个参与校区——巴吞鲁日的南方大学和路易斯安那州的格兰布林州立大学——所采用的研究方法。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

An idea that has gained support in recent years is that epigenetic changes to DNA and natural cellular modifications to proteins may effect stress-induced immune responses. One of the major consequences of cellular stress is the production and secretion of Danger-Associated Molecular Patterns (DAMPS). The DAMPs, including high-mobility group box protein 1 (HMGB1) and heat-shock protein 70 (HSP70), induce inflammation in mammals by interacting with pattern-recognition receptor proteins (proteins capable of recognizing DAMP molecules). Earlier studies provide evidence about the possible role of chemical modifications to proteins in the secretion of HMGB1 by stressed cells. We do not know, however, if these modifications also affect secretion of HSP70. In this project, the investigators will use both computational and cell biological approaches to measure chemical changes to both DNA and proteins responsible for the release of DAMPs from lung and liver cells challenged with pentachlorophenol, a widely used organochlorine herbicide and wood preservative. The incorporation of computational approaches to understand these basic cellular mechanisms will greatly improve the research approach employed at the participating HBCU Campuses, Southern University in Baton Rouge and Grambling State University, both in Louisiana. An important highlight of this project is to increase the number of undergraduate African-American students entering careers as research scientists by providing these HBCU students with faculty-supervised training from qualified and trained HBCU Ph.D. students, which will also develop the mentoring abilities of the latter. This Excellence in Research award will enable the investigators to develop and establish a rigorous and high-quality research program that will significantly improve the research experience and education of minority students in Louisiana.The proposal investigates the molecular mechanisms associated with the endogenous expression and extracellular release of Danger-Associated Molecular Patterns (DAMPs) in cellular models under stressed conditions. The DAMPs include high-mobility group box protein 1 (HMGB1) and heat-shock protein 70 (HSP70). These proteins induce inflammation through interaction with cell surface or cytosolic pattern recognition receptors. Earlier studies provided evidence about the possible role of post-translational modification (PTM) in the secretion of HMGB1 by stressed cells, but little is known about HSP70’s secretory mechanisms. The investigators on this project will use both computational and cell biological approaches to identify the epigenetic factors and PTMs responsible for the release of DAMPs from lung and liver cells challenged with pentachlorophenol, an organochlorine herbicide and wood preservative. This will be accomplished in part through measurement of epigenetic methylation of DNA on CpG sequences in promoter regions of genes encoding DAMPs, as well as measurements of histone modifications in those same promoter regions. The additional incorporation of computational approaches to understand the basic cellular mechanisms will transform the research approaches being employed at the two participating HBCU Campuses, Southern University in Baton Rouge and Grambling State University, both in Louisiana.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.