INHIBITION OF MONOCYTE TGFB1-INDUCED SKIN FIBROSIS

抑制单核细胞 TGFB1 诱导的皮肤纤维化

• 批准号: 6045340
• 负责人: ANITA C GILLIAM
• 金额: $ 12.2万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-07-14 至 2003-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6045340
• 关键词: RNase protection assay; cellular immunity; flow cytometry; gene therapy; genetically modified animals; graft versus host disease; humoral immunity; in situ hybridization; laboratory mouse; migration inhibition factor; monocyte; pathologic process; scleroderma; sclerosis; transcription factor; transforming growth factors

The candidate, Anita C. Gilliam, is a junior faculty member on tenure track in Dermatology at Case Western Reserve University (CWRU), where she is developing a research career in molecular mechanisms of autoimmune disease. The proposed work draws on her experience in molecular biology and cutaneous immunobiology, and requests support for a mentored Clinical Scientist Development Award to acquire new expertise in monocyte biology under the mentorship of Dr. Kevin D. Cooper (Dermatology). The research environment, resources, and opportunities for career development at CWRU are superb, with CWRU School of Medicine recently listed as one of the top 10 research institutions in the country, and the Department of Dermatology as the top US program in NIH funding. The candidate's immediate goals are to develop the animal model in the proposed work into a vehicle useful for testing of interventions in scleroderma; long term goals are to develop the science of cutaneous monocyte biology and gene transfer in autoimmune disease as an independently funded investigator in an outstanding research environment. The proposal involves the study of systemic sclerosis/scleroderma, a chronic autoimmune disease of unknown etiology characterized by altered humoral and cell-mediated immunity, and excessive deposition of collagen in viscerae and skin, which is thought to be driven by activated monocytes making TGFbeta. We have characterized a very promising murine model for scleroderma that recapitulates many important features of scleroderma. Hypothesis: Monocytes are critical effector cells in Scl GVHD. TGF -beta1 is a major fibrogenic cytokine driving skin fibrosis in mice with Scl GVHD; TGF-beta2 and TGF-beta3 play minor if any roles in this cutaneous fibrosis. Skin fibrosis can be inhibited by: 1) antibodies to TGF-beta, 2), and latency associated peptide (LAP), a naturally occurring antagonist for TGF-beta. Specific Aim 1: To investigate the mechanism and sequence of early critical events leading to skin fibrosis in animals with Scl GVHD to better understand the pathophysiology of fibrosing disease and to devise novel focused interventions. Establishing the critical parameters of monocyte influx into skin, production of fibrogenic TGFbeta, and upregulation of proalpha(I) collagen mRNA synthesis provides a foundation for in vivo focused interventions. Specific Aim II: To further characterize in vivo interventions that inhibit TGFbeta1 in Scl GVHD. We have shown in preliminary experiments that TGF-beta LAP and antibodies to TGF-beta inhibit skin fibrosis in animals with Scl GVHD. Further characterization of these observations have high relevance to our understanding of basic monocyte biology, to the TGFbeta-driven fibrosing process in the animal model and in the autoimmune disease scleroderma, and potentially to the treatment of scleroderma and human graft versus host disease.

候选人安妮塔·C·乔里安是凯斯西储大学(CWRU)皮肤科终身教职的初级教员，在那里她正在发展自身免疫性疾病分子机制的研究生涯。这项拟议的工作借鉴了她在分子生物学和皮肤免疫生物学方面的经验，并要求支持临床科学家发展导师奖，以在凯文·D·库珀博士(皮肤科)的指导下获得单核细胞生物学方面的新专业知识。CWRU的研究环境、资源和职业发展机会一流，CWRU医学院最近被列为全国十大研究机构之一，皮肤科被NIH资助的美国项目中排名第一。候选人的近期目标是将拟议工作中的动物模型发展成一种可用于硬皮病干预测试的工具；长期目标是发展皮肤单核细胞生物学和自身免疫性疾病中的基因转移科学，作为一名独立资助的研究员，在出色的研究环境中。该提案涉及对系统性硬化症/硬皮病的研究，这是一种病因不明的慢性自身免疫性疾病，其特征是体液和细胞介导的免疫功能改变，以及内脏和皮肤中胶原的过度沉积，这被认为是由激活的单核细胞产生TGFbeta推动的。我们已经确定了一种非常有希望的硬皮病小鼠模型，该模型概括了硬皮病的许多重要特征。假设：单核细胞是SCL GVHD中的关键效应细胞。在慢性移植物抗宿主病小鼠中，转化生长因子-β1是导致皮肤纤维化的主要细胞因子；转化生长因子-β2和转化生长因子-β3在皮肤纤维化中的作用很小。皮肤纤维化可以通过：1)抗转化生长因子-β抗体，2)和潜伏期相关肽(LAP)，一种自然产生的转化生长因子-β拮抗剂来抑制。具体目标1：研究SCL GVHD动物早期导致皮肤纤维化的关键事件的机制和顺序，以更好地了解纤维化疾病的病理生理学机制，并设计新的有针对性的干预措施。建立单核细胞进入皮肤、产生致纤维化的转化生长因子β和上调前α(I)胶原基因合成的关键参数，为体内有针对性的干预提供了基础。特定目的II：进一步确定抑制SCL GVHD中TGFbeta1的体内干预措施的特征。我们已经在初步实验中表明，转化生长因子-βLAP和转化生长因子-β抗体可抑制SCL-GVHD动物的皮肤纤维化。这些观察结果的进一步表征与我们对基础单核细胞生物学的理解高度相关，与转化生长因子β驱动的动物模型和自身免疫性疾病硬皮病的纤维化过程密切相关，并可能与硬皮病和人类移植物抗宿主病的治疗有关。