Adhesion Junction Regulation in Normal and Diseased Skin

正常和患病皮肤的粘附连接调节

基本信息

批准号：
8524144
负责人：
DAVID S RUBENSTEIN
金额：
$ 40.91万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-07-01 至 2014-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8524144
关键词：
Acantholysis Actins Adhesions Autoantibodies Autoimmune Process Autoimmune Responses Binding Biochemical Biological Biological Process Biology Bulla Cell membrane Cell-Cell Adhesion Cells Characteristics Clinic Clinical Clinical Trials Design Combination Drug Therapy Complex Cytoskeleton Desmosomes Disease Dissociation Drug usage Early Endosome Endocytosis Epidermal Growth Factor Receptor Epithelial Epithelium Event Family Funding Genetic Goals Grant Heat Shock Protein 27 Histologic Human Immune response Immunoglobulin G Immunosuppressive Agents In Vitro Intermediate Filaments Keratin Laboratories Leucine Zippers Life MAPK14 gene MAPK8 gene Mediating Mitogen-Activated Protein Kinases Modeling Molecular Mucous Membrane Mus Neonatal Oncogenes Organ Outcome Study Pathogenesis Pathway interactions Patients Pemphigus Pemphigus Vulgaris Pharmaceutical Preparations Phosphorylation Physiological Physiology Process Protein Kinase Proteins Regulation Role Secondary to Series Serum Signal Transduction Skin Steroids Testing Toxic effect Translating Variant Wound Healing computerized data processing desmocollin desmoglein desmoglein III desmoplakin effective therapy efficacy testing human MAPK14 protein human disease in vitro testing in vivo inhibitor/antagonist insight interest keratinocyte mortality novel strategies plakoglobin plakophilins skin disorder therapy development

项目摘要

DESCRIPTION (provided by applicant): We are interested in understanding (1) how cells regulate their ability to adhere to one another, (2) how cell- cell adhesion complexes might also function to transduce signals that inform the cell about the state of adhesion, a process known as signal transduction, and (3) the relationship of these regulatory events to human skin disease. This application represents the competitive continuation for this grant, which is funded through June 2012. Study of the mechanism by which PV IgG induces loss of adhesion in target keratinocytes has enabled us to build a model of desmosome regulation and signaling in which PV IgG bind to desmoglein 3 on the keratinocyte cell membrane and initiate a series of biochemical events including phosphorylation of p38 mitogen activated protein kinase (p38MAPK, p38) and heat shock protein 27 (HSP27). Funding of this grant over the past 4 years has enabled us to further elucidate the mechanism by which pemphigus IgG induce loss of adhesion in target keratinocytes. We have identified additional regulatory components of this mechanism enabling us to build a more integrated model of pathogenesis. Events downstream of p38 activation include (i) EGFR activation/phosphorylation, (ii) endocytosis of dsg3, plakoglobin and EGFR into early endosomes, (iii) and HSP27 phosphorylation. This process culminates in the destabilization of desmosomes, retraction of keratin intermediate filaments and alterations to the actin cytoskeleton, which collectively contribute to the loss of cell-cell adhesion/acantholysis. Additionally, we have attempted to translate our observations from the laboratory to the clinic through a clinical trial designed to test the efficacy of a proprietary orl p38 inhibitor in patients with pemphigus vulgaris. We hypothesize that the signaling and intracellular biochemical changes induced by PV IgG are an exaggeration of normal physiology that provides insight into the regulation of desmosome mediated cell-cell adhesion. In this proposal, we outline our plan to (i) further elucidate the intracellular signaling processes and mechanism of pemphigus acantholysis, (ii) to extend our understanding of desmosome regulation by pemphigus IgG to other pathophysiologic processes by determining if and characterizing these desmosome signaling and regulatory processes in biological events involving transitions in desmosome adhesion, and (iii) test the hypothesis in vitro and in vivo if targeting multiple components of the pathway simultaneously with a combination of different drugs can inhibit acantholyis with the goal to minimize drug concentrations when used in combination and therefore minimize potential off-target effects and systemic toxicities compared to when these drugs are used individually. As such, a significant outcome of these studies will be to translate our observations into effective treatments for pemphigus.

描述（由申请人提供）：我们有兴趣了解（1）细胞如何调节其相互粘附的能力，（2）细胞-细胞粘附复合物如何也可以发挥作用来转导信号，告知细胞粘附状态，一个称为信号转导的过程，以及（3）这些调节事件与人类皮肤病的关系。该申请代表了该资助的竞争性延续，该资助截止至 2012 年 6 月。对 PV IgG 诱导目标角质形成细胞粘附丧失的机制的研究使我们能够建立桥粒调节和信号传导模型，其中 PV IgG 与桥粒芯糖蛋白 3 位于角质形成细胞的细胞膜上，并启动一系列生化事件，包括 p38 丝裂原激活蛋白激酶 (p38MAPK、p38) 和热休克蛋白的磷酸化27 (HSP27)。过去 4 年的资助使我们能够进一步阐明天疱疮 IgG 诱导目标角质形成细胞粘附丧失的机制。我们已经确定了该机制的其他调节成分，使我们能够建立更加综合的发病机制模型。 p38 激活的下游事件包括 (i) EGFR 激活/磷酸化，(ii) dsg3、斑珠蛋白和 EGFR 内吞到早期内体中，(iii) 和 HSP27 磷酸化。这一过程最终导致桥粒不稳定、角蛋白中间丝回缩和肌动蛋白细胞骨架改变，这些共同导致细胞间粘附/棘层松解的丧失。此外，我们还尝试通过一项临床试验将我们的实验室观察结果转化为临床，该临床试验旨在测试专有的 orl p38 抑制剂对寻常型天疱疮患者的疗效。我们假设 PV IgG 诱导的信号传导和细胞内生化变化是正常生理学的夸大，这为了解桥粒介导的细胞间粘附的调节提供了见解。在本提案中，我们概述了我们的计划：(i) 进一步阐明天疱疮棘层松解症的细胞内信号传导过程和机制，(ii) 通过确定和表征这些桥粒信号传导和特征，将我们对天疱疮 IgG 桥粒调节的理解扩展到其他病理生理过程。涉及桥粒粘附转变的生物事件中的调节过程，以及（iii）在体外和体内测试假设，如果同时使用不同药物的组合靶向该途径的多个成分可以抑制acantholyis 的目标是在联合使用时最大限度地降低药物浓度，从而与这些药物单独使用时相比，最大限度地减少潜在的脱靶效应和全身毒性。因此，这些研究的一个重要成果是将我们的观察结果转化为天疱疮的有效治疗方法。