Keratin Biology and Disease Aspects in Simple Epithelia

单上皮细胞的角蛋白生物学和疾病方面

• 批准号: 8424315
• 负责人: Bishr Omary
• 金额: $ 42.62万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-02-01 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8424315
• 关键词: Abbreviations; Acetylglucosamine; Actins; Acute; Acute Liver Failure; Animal Model; Antibodies; Apoptosis; Apoptotic; Bacteria; Binding; Biochemical; Biological Models; Biology; Blood Circulation; Bulla; Caspase; Cell Culture Techniques; Cell surface; Cells; Cleaved cell; Colitis; Colon; Coupled; Cytoskeletal Proteins; Detection; Diagnostic; Digestion; Digestive System Disorders; Disease; Disease Association; Epithelium; Exocrine pancreas; Extravasation; Family; Gallbladder; Generations; Genes; Genetic Models; Goals; Health; Hepatobiliary; Hepatocyte; Housing; Hyperplasia; Inflammatory Bowel Diseases; Intermediate Filament Proteins; Intermediate Filaments; Intestines; K-18 conjugate; KRT19 gene; Keratin; Knockout Mice; Knowledge; Link; Liver; Liver diseases; Microbe; Microfilaments; Microtubules; Molecular; Molecular Profiling; Mutation; Myosin ATPase; Myosin Heavy Chains; Myosin Light Chains; O-GlcNAc transferase; Okadaic Acid; Organ; Patients; Phenotype; Phenylcarbamates; Phosphorylation; Physiological; Physiology; Play; Prognostic Marker; Protein Family; Proteins; Regulation; Risk Factors; Role; Simple Epithelium; Site; Streptozocin; System; Tissue Polypeptide Specific Antigen; Toll-like receptors; Transgenic Animals; Tubulin; cholangiocyte; disease-causing mutation; glycosylation; human disease; liver injury; member; non-muscle myosin; novel; peptide O-linked N-acetylglucosamine-beta-N-acetylglucosaminidase; polypeptide; prognostic; protein function; response; skin disorder; translational study

DESCRIPTION (provided by applicant): The overall goal of our studies is to understand the regulation, function, and disease association of the keratin intermediate filament (IF) proteins, keratin polypeptides 8, 18 and 19 (K8, K18 and K19). These keratins are the major IF proteins in epithelia of digestive organs. The functions of keratins are becoming increasingly understood and their significance is highlighted by their growing association with a broad range of human diseases. For example, several skin diseases are caused by mutations in epidermal keratins, and K8 and K18 mutations predispose to liver disease and its progression. Our overall hypothesis is that IF proteins, one of the three major cytoskeletal protein families that include actin microfilaments and tubulin microtubules, play important physiologic and disease-related roles whose significance is rapidly unfolding. We propose to use two approaches: (a) a targeted approach to understand specific aspects of keratin regulation; namely glycosylation, keratin associated proteins (KAPs), and keratin cleavage by caspases, and (b) an unbiased approach to explore results from expression profiling analysis of wild-type and keratin-8 null mouse colonocytes to understand the consequent colitis and colonic hyperplasia that are due to keratin absence. Our specific aims are: (i) Study the functional significance of keratin glycosylation in digestive organs. The hypothesis for Aim#1 is that keratin glycosylation plays an important role in regulating the phosphorylation of other essential cellular proteins. (ii) Characterize the interaction and function of the association between keratins and non-muscle myosin. The hypothesis for Aim#2 is that KAPs regulate keratin function and that KAP function may be regulated by keratins. (iii) Examine the utility of assessing keratin apoptotic fragments in patients with acute liver failure. The hypothesis for Aim#3 is that apoptosis during acute liver injury results in leakage of the highly abundant keratin caspase-cleaved fragments into the circulation and that the presence of such fragments will have diagnostic and/or prognostic implications. (iv) Understand how keratins and luminal microbes modulate colonic hyperproliferation via apoptotic effects. The hypothesis for Aim#4 is that changes in colonocyte gene and cell surface expression in response to keratin absence, coupled with the presence of luminal bacteria, inhibit apoptosis. Our proposal includes cell biologic, molecular, cell culture, genetic models and human disease-related studies aimed at helping understand the regulation, function and disease aspects of keratins in digestive organs. Fundamental findings that we hope to show include a function for keratin glycosylation, myosin as a novel phosphorylation-dependent keratin binding partner, keratin apoptotic fragments as diagnostic or prognostic markers in liver disease, and an unexpected role of keratins in the colon as proapoptotic proteins as contrasted with their role in the liver as anti-apoptotic.

描述（由申请人提供）：我们研究的总体目标是了解角蛋白中间丝（IF）蛋白，角蛋白多肽8、18和19（K8，K18和K19）的调节，功能和疾病关联。这些角蛋白是消化器官上皮中的蛋白质的主要IF。角蛋白的功能越来越众所周知，它们与广泛的人类疾病的关联增强了它们的意义。例如，几种皮肤疾病是由表皮角蛋白突变引起的，而K8和K18突变易感肝病及其进展。我们的总体假设是，如果蛋白质是包括肌动蛋白微丝和微管蛋白微管的三个主要细胞骨架蛋白家族之一，则扮演重要的生理和疾病相关角色，其意义正在迅速发展。我们建议使用两种方法：（a）一种有针对性的方法来了解角蛋白调节的特定方面；即胱天蛋白酶酶的糖基化，角蛋白相关的蛋白（KAPS）和角蛋白裂解，以及（b）一种无偏见的方法来探索野生型和角蛋白-8零小鼠结肠的表达分析分析的结果，以了解结果的结肠炎和结肠炎，是伴有伴蛋白蛋白的结果。我们的具体目的是：（i）研究消化器官中角蛋白糖基化的功能意义。 AIM＃1的假设是角蛋白糖基化在调节其他必需细胞蛋白的磷酸化方面起着重要作用。 （ii）表征角蛋白与非肌肉肌球蛋白之间关联的相互作用和功能。 AIM＃2的假设是KAPS调节角蛋白功能，而KAP功能可以由角蛋白调节。 （iii）检查评估急性肝衰竭患者角蛋白凋亡片段的实用性。 AIM＃3的假设是，急性肝损伤期间的凋亡导致高度丰富的角蛋白caspase裂解的片段泄漏到循环中，并且这种片段的存在将具有诊断和/或预后的含义。 （iv）了解角蛋白和腔微生物如何通过凋亡作用调节结肠高增殖。 AIM＃4的假设是，对角蛋白的缺乏，结肠细胞基因和细胞表面表达的变化以及腔细菌的存在，抑制了凋亡。我们的建议包括细胞生物学，分子，细胞培养，遗传模型和与人类疾病有关的研究，旨在帮助了解消化器官中角蛋白的调节，功能和疾病方面。我们希望表现出的基本发现包括角蛋白糖基化的功能，肌球蛋白作为一种新型磷酸化依赖性角蛋白结合伴侣，角质蛋白凋亡片段，作为诊断或预后标记在肝病中的诊断或预后标记，以及角质素在Colon中的意外作用，作为对比型的角色，因为它们与其在对比中的作用。