Differentiating Ulcerative Colitis and Crohns Colitis Through Proteomic Patterns

描述（由申请人提供）：虽然克罗恩结肠炎（CC）和溃疡性结肠炎（UC）有几个共同的临床特征，但它们不同，有不同的病因和组织损伤的离散机制，治疗方案也有很大不同。因此，准确诊断炎症性肠病（IBD）在医疗护理、手术干预和预后方面具有至关重要的意义。UC和CC的区别是基于临床、放射学、内窥镜和病理解释，但在多达15%的IBD患者中无法区分。这被称为不确定性结肠炎（IC）。约90%的IC在因暴发性结肠炎而行结肠切除术时被诊断出来，随后的治疗严重依赖于正确的诊断。我们已经开发了一种适用的蛋白质组学方法，支持诊断的可行性，以区分分子，不同的炎性结肠炎。以往的研究在血清、粪便和粘膜活检生物标志物研究中都取得了不同程度的成功，以区分预后活动性疾病和静止状态。然而，这些生物标志物在CC和UC之间没有区别。专门的基质辅助激光解吸/电离质谱（MALDI MS）提供了直接对组织进行蛋白质组学生物标志物评估的可能性。在我们的初步研究中，使用MALDI质谱分析来自确诊UC、CC、IC、结肠癌和相关非炎症正常组织（作为对照）的患者的结肠切除术样本的组织学层，进行蛋白质组学分析。结果成功地识别了11个非常重要的质量电荷比（m/z）信号，这些信号将UC与CC和正常对照区分开。这些m/z值显示所有这些特征在CC中都显示出更大的折叠诱导。其中一些特征仅在结肠粘膜下层发现，而另一些特征在粘膜和粘膜下层都发现。在R21基金的资助下，在目标1中，我们将使用质谱技术识别11个具有统计意义的指纹。第二个目的是通过免疫组化、免疫印迹和/或酶联免疫吸附试验验证所鉴定的蛋白在结肠组织层中的存在。这将允许追求进一步的资金（R01），这将允许

DESCRIPTION (provided by applicant): Although Crohn's colitis (CC) and ulcerative colitis (UC) share several clinical features they are different and have different causes and discrete mechanisms of tissue damage and treatment options differ significantly. Therefore the accurate diagnosis of inflammatory bowel disease (IBD) is of paramount importance in terms of medical care, surgical intervention and prognosis. The distinction between UC and CC is made on the basis of clinical, radiologic, endoscopic, and pathologic interpretations, but cannot be differentiated in up to 15% of IBD patients. This is called indeterminate colitis (IC). About 90% o IC is diagnosed at the time of colectomy for fulminant colitis and subsequent management critically depends on the correct diagnosis. We have developed an amenable proteomic methodology that supports the diagnostic feasibility to discriminate molecularly, different inflammatory colitis. Previous research has been variably successful in serum, in stool and in mucosal biopsy biomarker studies to differentiate prognostically active disease and quiescent state. These biomarkers however, were not discriminatory between CC and UC. Specialized matrix assisted laser desorption/ionization mass spectrometry (MALDI MS) offers the possibility of proteomic biomarker assessment of the tissue directly. In our preliminary studies, the histologic layers of colectomy samples from patients with confirmed UC, CC, IC, colon cancer, and associated non inflamed normal tissue used as controls) were analyzed using MALDI MS for proteomic profiling. The results have successfully identified 11 highly significant mass-to-charge ratio (m/z) signals that distinguish UC from CC and from normal controls. These m/z values displays all of which showed greater fold induction in CC. Some of these signatures were only found in the colonic submucosa while others were found in both the mucosa and submucosa. With the R21 funding, in Aim 1, we would like to identify the 11 statistically significant finger prints using Mass spectrometry cutting-edge technology. The second aim will be to validate the presence of the identified proteins in the colon tissue layers using IHC, Western blot and/ or ELISA assays. This will allow pursuit of further funding (R01) that will allow identification of serum-based markers found and validated in aims 1 and 2 for colitis-specific fingerprint in serum, and eventually develop antibodies for the novel proteins with no available immunoreagents in the market. This will also allow testing our hypothesis of delineating IC into either UC or CC through identifying signatures in endoscopic tissue samples from patients with colitis as well as eventually creating a serum biomarker assay to delineate the inflammatory colitides using same protein(s). This information may provide new avenues for the development of novel diagnostic, prognostic and therapeutic targets.