Differentiating Ulcerative Colitis and Crohns Colitis Through Proteomic Patterns

通过蛋白质组模式区分溃疡性结肠炎和克罗恩斯结肠炎

• 批准号: 8445993
• 负责人: Amosy Ephreim M'Koma
• 金额: $ 22.71万
• 依托单位: MEHARRY MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8445993
• 关键词: 3-hydroxy-3-methylglutaryl-coenzyme A; Anastomosis - action; Antibodies; Anus; Biological; Biological Assay; Biological Markers; Biometry; Biopsy; Caring; Categories; Charge; Classification; Clinical; Clinical/Radiologic; Colectomy; Colitis; Collaborations; Colon; Colon Carcinoma; Complication; Crohn' s disease; Development; Diagnosis; Diagnostic; Disease; Enzyme-Linked Immunosorbent Assay; Excision; Failure; Feces; Fingerprint; Fingers; Funding; Histologic; Ileal Reservoirs; Ileostomy; Immunohistochemistry; Individual; Inflammatory; Inflammatory Bowel Diseases; Inflammatory disease of the intestine; Interleukin-7; Lactoferrin; Lead; Leukocyte Elastase; Leukocyte L1 Antigen Complex; Marketing; Mass Spectrum Analysis; Medical; Methodology; Molecular; Monoclonal Antibodies; Mucous Membrane; Natural History; Normal tissue morphology; Operative Surgical Procedures; Oryctolagus cuniculus; Outcome; Oxidoreductase; Parents; Pathologic; Pathology; Patients; Pattern; Peptides; Phenotype; Placental Growth Factor; Printing; Prognostic Marker; Protein Databases; Proteins; Proteome; Proteomics; Research; S100A12 gene; Sampling; Sequence Analysis; Serum; Signal Transduction; Specimen; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Submucosa; Surgeon; Technology; Testing; Time; Tissue Sample; Tissues; Ulcerative Colitis; Western Blotting; base; density; desmoglein; diagnostic accuracy; disease classification; experience; falls; improved; insight; interest; interleukin-12 subunit p40; large bowel Crohn' s disease; liquid chromatography mass spectrometry; novel; novel diagnostics; outcome forecast; overexpression; platelet protein P47; prognostic; public health relevance; response; rho; therapeutic target

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.

描述（由申请人提供）：尽管克罗恩氏结肠炎（CC）和溃疡性结肠炎（UC）有几个共同的临床特征，但它们是不同的，具有不同的病因和组织损伤的离散机制，治疗选择也有显著差异。因此，炎症性肠病（IBD）的准确诊断在医疗护理、外科干预和预后方面至关重要。UC和CC之间的区别是基于临床，放射学，内窥镜和病理学解释，但在高达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在结肠切除术时被诊断为暴发性结肠炎，随后的治疗关键取决于正确的诊断。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，不同的炎症性结肠炎。先前的研究已经在血清、粪便和粘膜活检生物标志物研究中成功地区分了活动性疾病和静止状态。然而，这些生物标志物不能区分CC和UC。专业的基质辅助激光解吸/电离质谱（MALDI MS）提供了直接评估组织蛋白质组生物标志物的可能性。在我们的初步研究中，使用MALDI MS分析了来自确诊为UC、CC、IC、结肠癌的患者的结肠切除术样品的组织学层，以及用作对照的相关非炎症正常组织。结果成功地确定了11个高度显着的质荷比（m/z）信号，区分UC从CC和正常对照。这些m/z值显示所有这些在CC中显示更大的倍数诱导。这些特征中的一些仅在结肠粘膜下层中发现，而另一些在粘膜和粘膜下层中发现。有了R21的资助，在目标1中，我们希望使用质谱尖端技术识别11个具有统计学意义的指纹。第二个目的是使用IHC、Western印迹和/或ELISA测定来验证结肠组织层中所鉴定的蛋白质的存在。这将允许寻求进一步的资金（R 01）， 鉴定在目标1和2中发现并验证的血清标记物，以确定血清中结肠炎特异性指纹，并最终在市场上没有可用免疫试剂的情况下开发新蛋白质的抗体。这也将允许通过识别结肠炎患者的内窥镜组织样本中的特征以及最终创建血清生物标志物测定以使用相同的蛋白质描绘炎性结肠炎来测试我们将IC描绘为UC或CC的假设。这些信息可能为开发新的诊断、预后和治疗靶点提供新的途径。