Predictive biomarkers for EAE development

描述(申请人提供)：大约千分之一的人患上多发性硬化症，但即使在表达多发性硬化症相关的MHC II等位基因的人中，我们也无法预测谁会患上疾病，而且只有千分之一的人会得多发性硬化症。令人惊讶的是，即使在多发性硬化症的小鼠实验性自身免疫性脑脊髓炎(EAE)模型的研究中，在明确定义的条件下诱发主动或被动疾病，但并不是所有基因相同的小鼠都会患上相同的疾病轨迹或严重程度，我们也无法准确预测谁会患上疾病。在我们自己的初步研究中，我们已经使用我们最近报道的一种新的定量蛋白质组学方法确定了几个与EAE的发病、高峰和缓解相关的蛋白质生物标志物候选(Raphael，I.Electric，2012)。此外，我们开发了一种方法，使用过继转移EAE结合转移前细胞因子ELISPOT分析来诱导预定严重程度的EAE。重要的是，我们已经有了初步的结果，可以用ELISA法在EAE小鼠的血清中检测到预测的候选蛋白质生物标志物。通过结合这些诱发EAE的方法和将蛋白质表达与疾病进展相关联，我们将在一个独特的系统中提供原则证明，以确定疾病发病率和严重性的预测标记。这项提案的目的是利用EAE模型提供预测生物标记物的原则证明。我们的中心假设是，在临床上出现EAE症状之前，中枢神经系统疾病特异性蛋白会释放到血液(血清)中，这种蛋白可以用来预测哪种动物会发展成疾病以及病情有多严重。这一假设是基于我们自己的初步结果，并得到了已发表的研究的支持。提出这项研究的基本原理是，EAE模型中的原则证明将为开发MS患者的同源预测生物标志物提供基础，这将为MS的治疗和药物开发带来革命性的变化。我们将以以下具体目标来检验我们的中心假设：目的1.确定在EAE发病前中枢神经系统疾病相关蛋白亚型的关键表达变化，并作为疾病严重程度的函数。目的2.确定最能预测EAE发病和严重程度的CNS疾病相关蛋白亚型释放到血液(血清)。我们预计，拟议的研究将提供目前无法获得的重要信息，并将对MS的研究和治疗产生重大和持久的影响。

DESCRIPTION (provided by applicant): Approximately 1/1000 people develop multiple sclerosis (MS), but even among people that express the MS- associated MHC II allele HLA-DR2 we cannot predict who will develop disease and only 1/200 HLA-DR2+ people ever will. Surprisingly, even in studies of the murine experimental autoimmune encephalomyelitis (EAE) model of MS, where active or passive disease is induced under well-defined conditions, not all of the genetically-identical mice develop disease with the same trajectory or severity and we cannot accurately predict who will develop disease. In our own preliminary studies we have identified several protein biomarker candidates that correlate with onset, peak, and remission of EAE using a novel quantitative proteomics method that we recently reported (Raphael, I. Electrophoresis, 2012). Furthermore, we have developed a method to induce EAE of predetermined severity using adoptive transfer EAE in combination with pre-transfer cytokine ELISPOT analysis. Importantly, we have preliminary results that predicted candidate protein biomarkers can be detected in serum of mice with EAE by ELISA. By combining these approaches for inducing EAE and correlating protein expression to disease progression we will provide proof-of-principle in a unique system to identifying predictive markers of disease incidence and severity. The objective of this proposal is to provide proof-of-principle of predictive biomarkers using the EAE model. Our central hypothesis is that the clinical onset of EAE symptoms is preceded by release of CNS disease- specific proteins into blood (serum) that can be used to predict which animal will develop disease and how severe. This hypothesis is based on our own preliminary results and supported by published studies. The rationale for the proposed research is that proof-of-principle in the EAE model will provide the basis for developing homologous predictive biomarkers for MS patients, which will revolutionize treatment and drug development for MS. We will test our central hypothesis with the following specific aims: Aim 1. To determine key CNS disease-related protein isoforms with altered expression in CNS tissue prior to the onset of EAE and as a function of disease severity. Aim 2. To determine the CNS disease-related proteins isoforms released into blood (serum) most predictive of EAE incidence and severity. We expect that the proposed studies will provide important information that is currently not available and that will have significant and lasting impact on research and treatment of MS.