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

近年来获得支持的一个想法是，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。 DAMP，包括高迁移率族蛋白1（HMGB1）和热休克蛋白70（HSP 70），通过与模式识别受体蛋白（能够识别DAMP分子的蛋白质）相互作用诱导哺乳动物的炎症。 早期的研究提供了有关蛋白质化学修饰在应激细胞分泌HMGB1中可能作用的证据。 然而，我们不知道这些修饰是否也影响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DNA和蛋白质的化学变化，这些变化负责从肺和肝细胞释放DAMP，这些细胞受到五氯苯酚的挑战，五氯苯酚是一种广泛使用的有机氯除草剂和木材防腐剂。 结合计算方法来理解这些基本的细胞机制将大大提高参与HBCU校园，南方大学在巴吞鲁日和格兰布林州立大学，都在路易斯安那州采用的研究方法。 该项目的一个重要亮点是通过为这些HBCU学生提供来自合格和训练有素的HBCU博士的教师监督培训，增加进入研究科学家职业的本科非洲裔美国学生的数量。学生，这也将发展后者的辅导能力。 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和高质量的研究计划，这将显着提高路易斯安那州少数民族学生的研究经验和教育。该提案调查了与内源性表达和细胞外释放的危险相关分子模式（DAMP）在细胞模型在应激条件下的分子机制。 DAMPs包括高迁移率族蛋白1（HMGB1）和热休克蛋白70（HSP 70）。 这些蛋白质通过与细胞表面或胞质模式识别受体相互作用诱导炎症。 早期的研究证实了翻译后修饰（PTM）在应激细胞分泌HMGB1中的可能作用，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负责从肺和肝细胞释放DAMP的表观遗传因子和PTM，这些细胞受到五氯苯酚（一种有机氯除草剂和木材防腐剂）的挑战。这将部分地通过测量编码DAMP的基因的启动子区域中的CpG序列上的DNA的表观遗传甲基化以及测量那些相同启动子区域中的组蛋白修饰来实现。 通过计算方法来理解基本的细胞机制，将改变HBCU两个参与校园，巴吞鲁日的南方大学和路易斯安那州的Grambling州立大学的研究方法。该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估来支持。

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.