Mechanism of Keratinocyte Motility

角质形成细胞运动机制

• 批准号: 7227110
• 负责人: DAVID T. WOODLEY
• 金额: $ 34万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-03-24 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7227110
• 关键词: Actins; Acute; Address; Animal Model; Appendix; Area; Basement membrane; Bathing; Beds; Biological; Biological Assay; Blood Vessels; Bovine Pituitary Extract; California; Cell Communication; Cell Cycle Regulation; Cell secretion; Cells; Chicago; Clinical; Cloning; Closure; Coagulation Process; Collaborations; Collagen; Computer Assisted; Computer Systems Development; Condition; Connective Tissue; Data; Depth; Desmosomes; Doctor of Medicine; Dominant-Negative Mutation; Educational process of instructing; Elements; Engineering; Environment; Event; Extracellular Matrix; Fibroblasts; Fibronectins; Filopodia; Focal Adhesion Kinase 1; Focal Adhesions; Funding; Gelatinase A; Gelatinase B; Gene Expression; Genes; Genetic Medicine; Genetics and Medicine; Goals; Grant; Green Fluorescent Proteins; Growth Factor; Hand; Healed; Hemidesmosomes; Human; Human Engineering; Hypoxia; Immigration; Immunocompromised Host; In Vitro; Inbred HRS Mice; Infection; Institutes; Integrins; Investigation; Laboratory Research; Lateral; Lead; Left; Link; Literature; MAPK14 gene; MCDB 153; Malignant Neoplasms; Matrix Metalloproteinases; Mediating; Metalloproteases; Microscopy; Migration Assay; Mitogen Activated Protein Kinase 1; Modeling; Molecular; Mothers; Mus; Nature; Noise; Nori; Numbers; Occlusive Dressings; Oxygen measurement, partial pressure, arterial; Paper; Pathway interactions; Pattern; Pharmaceutical Preparations; Physiological; Plasma; Play; Principal Investigator; Process; Production; Proteins; Publishing; Reader; Recruitment Activity; Regulation; Research; Research Personnel; Role; Running; Series; Serum; Side; Signal Pathway; Signal Transduction; Site; Skin; Source; Sterile coverings; Stimulus; Stress; Suggestion; Surface; System; Technology; Testing; Time; Transcriptional Activation; Transfection; Transgenes; Universities; Up-Regulation; Work; Wound Healing; appendage; cell motility; concept; cytokine; design; experience; extracellular; ezrin; gene delivery system; gene function; healing; human MAPK14 protein; in vivo; keratinocyte; laminin-1; laminin-5; migration; mitogen-activated protein kinase p38; moesin; mutant; new technology; novel; novel strategies; professor; programs; radixin protein; receptor; research study; response; transduction efficiency; wound

DESCRIPTION (provided by applicant): Human keratinocyte (HK) motility is a critical event in the process of wound healing. The biological elements that directly influence HK migration are incompletely understood. The microenvironment of the healing skin wound dictates experiments be designed that directly examine elements of the wound environment and evaluate their influence. Our 3 independent, computer-assisted HK migration assays are able to dissect HK motility from HK proliferation. We demonstrated that the type of connective tissue components juxtaposed to the HK has a major influence upon HK migration. In the last cycle, our data support a paradigm in which extracellular matrices (ECMs) can initiate HK migration in the absence of growth factors (GFs), so-called "ECM-initiated" motility or haptotaxis. GFs cannot initiate HK motility. Selected GFs are needed to enhance and optimize ECM-initiated migration. Hypoxia also increases HK migration on ECMs. This is in accordance with human skin wounds treated by semi-permeable occlusive dressings which promote re-epithelialization but make the wound oxygen tension immeasurably low. Therefore, we have identified 3 separable biological elements that profoundly influence HK motility: ECMs, GFs and hypoxia. During the last cycle, we focused on how hypoxia promotes HK motility at the cellular level. We found that hypoxia did notjalter the HK integrin profile, but decreased HK-derived laminins 1 and 5 (2 motility "brakes") and increased the lamellipodia-associated proteins - ezrin, moesin and radixin. It also increased selected matrix metalloproteinases such as MMP-9. We also identified 4 important signaling pathways needed for HK motility--p38 MAP kinase, ERK 1/2, PKC delta and FAK-driven signaling. In this proposal, we wish to determine (i) if hypoxia-enhanced motility is mediated via one or more of the 4 pathways we've identified as necessary for HK migration and if hypoxia-enhanced motility is mediated via ECM-initiated, GF-enhanced motility or both and (ii) which signaling pathways are responsible for cellular, morpholological, structural, mechanisms needed for motility such as focal adhesion turnover, lamillipodia formation, and cell polarization. Lastly, in our Third Aim, we will develop a novel animal model to examine the veracity of our in vitro data. This model will consist of genetically engineered human skin equivalents grafted onto hairless mice.

描述（由申请人提供）： 人角质形成细胞（HK）的运动是伤口愈合过程中的一个关键事件。直接影响香港移民的生物因素还不完全清楚。愈合皮肤伤口的微环境要求设计实验，直接检查伤口环境的元素并评估其影响。我们的3个独立的，计算机辅助的HK迁移试验能够从HK增殖中分离HK运动性。我们证明了与HK并列的结缔组织成分的类型对HK迁移有重大影响。在最后一个周期中，我们的数据支持这样一个范例，即细胞外基质（ECM）可以在没有生长因子（GF）的情况下启动HK迁移，即所谓的“ECM启动”运动或趋触性。GF不能启动HK运动。需要选定GF来增强和优化ECM启动的迁移。缺氧也会增加ECM上的HK迁移。这与通过半渗透性封闭敷料治疗的人类皮肤伤口一致，所述半渗透性封闭敷料促进上皮再生但使伤口氧分压不可测量地低。因此，我们已经确定了3个可分离的生物元素，深刻影响香港运动：ECM，GF和缺氧。在上一个周期中，我们重点关注缺氧如何在细胞水平上促进HK运动。我们发现缺氧并没有改变HK整合素的分布，但减少了HK衍生的层粘连蛋白1和5（2个运动性“刹车”），并增加了板状脂质相关蛋白- ezrin，moesin和radixin。它还增加了选定的基质金属蛋白酶，如MMP-9。我们还确定了HK运动所需的4条重要信号通路--p38 MAP激酶、ERK 1/2、PKC δ和FAK驱动的信号通路。在这个提议中，我们希望确定（i）缺氧增强的运动性是否通过我们已经确定为HK迁移所必需的4种途径中的一种或多种介导，以及缺氧增强的运动性是否通过ECM启动的、GF增强的运动性或两者介导，以及（ii）哪些信号传导途径负责运动性所需的细胞、形态学、结构机制，如粘着斑转换、板状伪足形成、和细胞极化。最后，在我们的第三个目标中，我们将开发一种新的动物模型来检查我们体外数据的准确性。这个模型将由移植到无毛小鼠身上的基因工程人类皮肤等同物组成。