Identification of an IL-6 induced keratinocyte motogen

IL-6 诱导的角质形成细胞运动原的鉴定

• 批准号: 7227129
• 负责人: RANDLE Michael GALLUCCI
• 金额: $ 24.26万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7227129
• 关键词: Antisense Oligonucleotides; Biological Assay; Cells; Chromatography; Chronic; Coculture Techniques; Condition; Conditioned Culture Media; Control Animal; Cutaneous; Data; Dermal; Development; Disease; Epidermis; Epithelial; Fibroblasts; Fingerprint; Gel; Gene Expression; Genes; Genomics; Goals; Health Professional; Immigration; In Vitro; Individual; Interleukin-6; Ion-Exchange Chromatography Procedure; Lead; Length; Mediating; Mediator of activation protein; Messenger RNA; Migration Assay; Molecular Profiling; Mus; Neonatal; Personal Satisfaction; Play; Proteins; Proteomics; Psoriasis; Relative (related person); Research Personnel; Role; Skin; Skin Ulcer; Stimulation of Cell Proliferation; System; Time; Transgenic Mice; Two-Dimensional Gel Electrophoresis; United States; Wild Type Mouse; Wound Healing; cell motility; cytokine; design; in vitro Model; inhibitor/antagonist; keratinocyte; mass spectrometer; migration; neutralizing antibody; programs; research study; response; wound

DESCRIPTION (provided by applicant): In the United States, over 6 million individuals develop chronic skin ulcers annually. The augmentation of cutaneous wound healing has long been an elusive goal for health care professionals. Our previous studies indicate that interleukin-6 deficient transgenic mice (IL-6KO) display significantly delayed cutaneous wound healing compared to wild type control animals. While the role of IL-6 is well documented in disease conditions such as psoriasis, little is known about the role this cytokine might play in regenerative responses such as wound healing. To further describe the role of IL-6 in skin wound healing, an in vitro model was developed utilizing cultured epidermal keratinocyte and dermal fibroblast cells from neonatal IL-6KO mice. This system allows for the direct assessment of the effects of IL-6 on skin cells without the confounding presence of endogenous IL-6. Using this culture system we have found that IL-6 appears to significantly induce cell motility, in cultured keratinocytes. However, this effect appears to be indirect and requires co-culture with dermal fibroblasts. Preliminary gene array experiments do not indicate the induction of a secreted protein known to induce keratinocyte migration. In this application we propose to: 1) characterize and 2) identify the IL-6-induced dermal fibroblast produced migratory factor. To do this, further gene array experiments will be conducted with IL-6KO dermal fibroblasts exposed to IL-6, and epidermal keratinocytes exposed to IL-6/fibroblast conditioned media. IL- 6/fibroblast conditioned media will also be concentrated, and ion exchange chromatography will be performed. Isolated IL-6KO keratinocytes will be exposed to fractions collected from the chromatographic separations, and migratory potential will be assessed. Fractions that induce a migratory response will be subject to proteomic analysis utilizing 2D gel seperation and tryptic fingerprinting will be determined with a MALDI-TOF mass spectrometer. Once identified, the motogenic potential of migratory factor(s) will be assessed on isolated keratinocytes from IL-6KO and wild type mice. The results of these experiments will hopefully lead to the eventual development of a useful treatment for chronic wounds.

描述（由申请人提供）：