MOLECULAR MECHANISMS CONTRIBUTING TO GENDER DISPARITY IN MULTIPLE SCLEROSIS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 多发性硬化症（MS）影响大约40万美国人，其中三分之二是女性。 MS的原因仍然难以捉摸，但髓鞘蛋白可能是负责启动CNS炎症反应的自身抗原，性类固醇可能调节对MS的易感性。阐明这些激素影响小胶质细胞功能的机制，并了解重要的髓鞘特异性蛋白的调节将是至关重要的，在理解MS性别差异的基础。 该项目的目标是确定导致MS中性别偏见的分子机制。将通过观察iNOS的表达水平和调节来评估女性性类固醇对小胶质细胞功能调节的影响，iNOS是负责该疾病特征性髓鞘随后降解的炎症分子产生的关键酶。 将iNOS-荧光素酶构建体瞬时转染到小胶质细胞中以检查该途径，并且最近几个月在实验室中优化了用于这些转染的有效系统。 实验正在进行中，以评估iNOS表达的女性性类固醇添加到培养基中。 性类固醇对免疫反应的调节可能导致MS的性别差异，但潜在自身抗原的性别特异性表达介导对疾病的易感性也可能有贡献。 现有的数据表明，在小鼠睾丸的Leydig细胞中，Plp mRNA的表达编码在成熟CNS髓鞘中发现的最丰富的蛋白质，以前被认为仅在CNS中表达，在雌性对应物中没有外周表达。 我们已经获得了蛋白质印迹数据证实睾丸组织中的Plp蛋白的表达。 这种外周表达可能在雄性中建立保护性耐受。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Multiple sclerosis (MS) affects approximately 400,000 Americans, two-thirds of whom are female. The causes of MS remain elusive, but a myelin protein is likely to be the autoantigen responsible for initiating the inflammatory response in the CNS, and sex steroids may modulate susceptibility to MS. Elucidation of the mechanisms by which these hormones influence microglial function and an understanding of the regulation of important myelin-specific proteins will be crucial in understanding the basis of gender differences in MS. The goals of this project are to determine molecular mechanisms that result in a gender bias in MS. Effects of female sex steroids upon the regulation of microglia function will be assessed by looking at expression levels and regulation of iNOS, a key enzyme in the production of inflammatory molecules responsible for the subsequent degradation of myelin characteristic of the disease. Transient transfections of iNOS-luciferase constructs into microglial cells will be done to examine this pathway, and an efficient system for these transfections has been optimized in the lab in recent months. Experiments are underway to assess iNOS expression in response to female sex steroids added to culture media. Modulation of the immune response by sex steroids is likely to contribute to the gender disparity of MS, but gender-specific expression of potential autoantigens in mediating susceptibility to the disease may also contribute. Existing data demonstrate expression of Plp mRNA encoding the most abundant protein found in mature CNS myelin, previously thought to be expressed exclusively in the CNS, in Leydig cells of the testes of mice, with no peripheral expression in female counterparts. We have obtained Western blot data confirming expression of Plp protein in testicular tissue. It is possible this peripheral expression could establish protective tolerance in males.