The Role of Caveolin-1 in Stress-Induced Premature Sensescence

Caveolin-1 在应激诱导的过早衰老中的作用

• 批准号: 7484460
• 负责人: Janine Bartholomew
• 金额: $ 1.58万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-06 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7484460
• 关键词: Age; Aging; Binding; Biochemical; CDKN1A gene; Caveolae; Caveolins; Cell Aging; Cell Cycle Arrest; Cells; Condition; Cyclin-Dependent Kinase Inhibitor 2A; Data; Dependence; Dermal; Embryo; Environment; Fibroblasts; Ficusin; Free Radicals; Galactosidase; Homeostasis; Hydrogen Peroxide; In Vitro; Inflammation; Investigation; Knockout Mice; Laboratories; MDM2 gene; MDM2 gene; Mediating; Membrane; Molecular; Mus; Mutation; Organism; Oxidative Stress; PUVA Photochemotherapy; Pathway interactions; Phenotype; Prevention; Proteins; Psoralens; Research Design; Role; Signal Transduction; Skin; Skin Aging; Staining method; Stains; Stress; Structural Protein; TP53 gene; Techniques; Tertiary Protein Structure; Testing; Time; Tissues; Transcriptional Activation; Up-Regulation; age related; beta-Galactosidase; carcinogenesis; caveolin 1; cell transformation; design; in vivo; interest; novel therapeutics; oncoprotein p21; photoactivation; response; scaffold; senescence; therapeutic target

DESCRIPTION (provided by applicant): Emerging data reveals that cellular senescence in vivo is due to stress-induced premature senescence (SIPS). SIPS is involved in organismal aging, under both pathological and normal conditions, with studies showing that senescent cells have negative effects on overall organism homeostasis. Psoralen photoactivation (PUVA), a common free radical-producing treatment for many skin inflammation ailments, is known to promote premature senescence of dermal fibroblasts and aging of the skin. However, the molecular mechanism underlying PUVA-induced senescence/aging remains largely unknown. The long term objective of this study is to determine cellular mechanisms that may be involved in stress induced premature senescence in vivo. Specifically, this proposal is interested in determining the role of caveolin-1 during PUVA-induced SIPS. This proposal investigates the hypothesis that caveolin-1 expression is required for PUVA-induced SIPS of dermal fibroblasts in vitro and in vivo through activation of the p53 pathway after sequestration of MDM2 into caveolar membranes. The following specific aims are designed to explore this hypothesis: 1) Show that PUVA induces SIPS in dermal fibroblasts through sequestration of MDM2 into caveolae in vitro and 2) determine if in vivo PUVA-induced SIPS of dermal fibroblasts is dependent upon Cav-1. Research designs consist of deriving dermal fibroblasts from mice and treating with PUVA to induce SIPS. Using mutational analysis of MDM2, we plan to determine whether the protein's sequestration by Caveolin-1 is needed for SIPS. We will be using common biochemical techniques along with senescence associated beta galactosidase staining to assess SIPS. Additionally, we plan to treat mice with PUVA and examine their dermal layers for PUVA-induced SIPS. By using wildtype and Caveolin-1 knockout mice, it will be assessed whether Caveolin-1 is necessary for PUVA-induced SIPS. These data will elucidate molecular players related to both normal and pathogenic organismal aging. Additionally, because PUVA is known to increase likeliness of carcinogenesis while aging the skin, and senescent cells are believed to create an environment conducive for preneoplastic cell transformation, our investigations intend to propose caveolin-1 as a novel therapeutic target for the treatment/ prevention of aging and aging-related carcinogenesis.

描述（由申请人提供）：新数据显示，体内细胞衰老是由于应激诱导的早衰（SIPS）。SIPS在病理和正常条件下参与生物体衰老，研究表明衰老细胞对整体生物体内稳态具有负面影响。已知Peglien光活化（PUVA），一种用于许多皮肤炎症疾病的常见的产生自由基的治疗，促进真皮成纤维细胞的过早衰老和皮肤老化。然而，PUVA诱导的衰老/老化的分子机制在很大程度上仍然未知。本研究的长期目标是确定可能参与应激诱导的体内早衰的细胞机制。具体而言，该提案感兴趣的是确定窖蛋白-1在PUVA诱导的SIPS过程中的作用。 该提案研究了这样的假设，即小窝蛋白-1表达是体外和体内皮肤成纤维细胞PUVA诱导的SIPS所需的，通过在MDM 2隔离到小窝膜后激活p53途径。设计以下具体目的以探索该假设：1）显示PUVA通过在体外将MDM 2隔离到小窝中来诱导真皮成纤维细胞中的SIPS，和2）确定体内PUVA诱导的真皮成纤维细胞的SIPS是否依赖于Cav-1。研究设计包括从小鼠中获得真皮成纤维细胞并用PUVA处理以诱导SIPS。使用MDM 2的突变分析，我们计划确定是否需要通过小窝蛋白-1的蛋白质的螯合SIPS。我们将使用普通的生化技术沿着衰老相关的β半乳糖苷酶染色来评估SIPS。此外，我们计划用PUVA治疗小鼠，并检查它们的真皮层是否存在PUVA诱导的SIPS。通过使用野生型和小窝蛋白-1敲除小鼠，将评估小窝蛋白-1对于PUVA诱导的SIPS是否是必需的。这些数据将阐明与正常和致病性有机体衰老相关的分子参与者。此外，由于已知PUVA在老化皮肤的同时增加致癌的可能性，并且衰老细胞被认为创造了有利于癌前细胞转化的环境，因此我们的研究旨在提出小窝蛋白-1作为治疗/预防衰老和衰老相关致癌的新治疗靶点。