Integrin-dependent mechanism of T cell-mediated pulmonary fibrosis

T细胞介导的肺纤维化的整合素依赖性机制

• 批准号: 8195953
• 负责人: Irina G. Luzina
• 金额: --
• 依托单位: BALTIMORE VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2012-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8195953
• 关键词: Address; Adverse effects; Allergic inflammation; Animal Model; Animals; Antibodies; Area; Arteriosclerosis; Asthma; Autoimmune Diseases; Autoimmunity; Binding; Bleomycin; Bronchiolitis Obliterans; CCL18 gene; Cause of Death; Cell Communication; Cell Culture Techniques; Cell surface; Cells; Cessation of life; Characteristics; Chronic; Chronic Disease; Cicatrix; Coculture Techniques; Collagen; Connective Tissue; Cross-Sectional Studies; Data; Deposition; Development; Diffuse; Disease; Elderly; Environmental Exposure; Equilibrium; Extracellular Matrix; Extrinsic allergic alveolitis; Fibroblasts; Fibrosis; Future; Goals; Hamman-Rich syndrome; Health; Home environment; Homing; Human; Immune; In Vitro; Infiltration; Inflammation; Inflammatory; Injury; Integrin alphaVbeta3; Integrins; Interferon Type II; Interferons; Interstitial Lung Diseases; Kidney; Knowledge; Liver; Lung; Lung diseases; Lymphocyte; Mediating; Membrane; Military Personnel; Mining; Mission; Modeling; Nature; Organ; Pathologic; Pathologic Processes; Patient Care; Patients; Phenotype; Play; Pneumonia; Population; Process; Production; Pulmonary Fibrosis; Pulmonology; Radiation; Regulation; Research; Rheumatoid Arthritis; Role; Sarcoidosis; Scleroderma; Small Interfering RNA; Syndrome; Systemic disease; T-Lymphocyte; TNF gene; Tissues; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Veterans; Wound Healing; airway remodeling; attenuation; base; chemotherapy; cytokine; healthy volunteer; human EBAF protein; human TNF protein; human subject; in vivo Model; integrin alphaVbeta5; male; neutralizing antibody; novel; overexpression; public health relevance; pulmonary function; research study; wound

DESCRIPTION (provided by applicant): Pulmonary fibrosis is a frequent consequence of chronic inflammatory processes and environmental expo- sures. Available antifibrotic therapies are inefficient. Data suggest that T cells are commonly present in the lungs of patients in association with fibrosis and express integrins, and that these integrins may be a master switch that defines a profibrotic or antifibrotic phenotype of T cells. The Overall Goal of this study is to clarify the mechanistic role of integrin-expressing T cells in the pro- and antifibrotic regulation in the lungs. The Spe- cific Hypothesis of this study is that the expression and functional activity of integrins 1V23, 1V25, 22- containing and 1E-containing integrins, on pulmonary T cells defines their regulatory phenotype (TGF-2+ simi- lar to Th3 and Tregs acting through membrane-bound TGF-2), persistence in the lung, and stimulation of col- lagen production. We further hypothesize that pulmonary infiltration of T cells that do not express integrins leads to a decrease in active TGF-2, elevated expression of TNF-1 and IFN-3, and subsequently to attenuation of collagen production. The following Specific Aims will be addressed. 1. Determine whether elevated expres- sion of integrins on pulmonary T cells is associated with a TGF-2-associated regulatory phenotype of T cells and with lower pulmonary function, whereas infiltration of T lymphocytes not expressing integrins is associated with inflammatory phenotype and higher pulmonary function in patients with interstitial lung disease. 2. Deter- mine whether elevated expression levels and functional activity of integrins on pulmonary T lymphocytes in the CCL18 overexpression model in vivo define a) their prolonged persistence in the lungs, b) the increased pro- duction, activation, and cell surface binding of TGF-2, and c) collagen accumulation. Determine whether partial neutralization of bleomycin-induced collagen accumulation by CCL18 overexpression is dependent on de- creased expression of 1V-containing integrins on pulmonary T lymphocytes, their decreased expression of ac- tive TGF-2, and elevation of TNF-1 and IFN-3. Determine whether these integrin-mediated pro- and antifibrotic mechanisms can be therapeutically modulated to regulate collagen accumulation in the lungs of experimental animals. 3. Determine whether 1V-, 22-, or 1E-containing integrins facilitate cell-cell interactions between pul- monary T cell and primary fibroblasts in vitro, the increase in TGF-2 production and/or activation, cell surface binding of TGF-2 by T cells, and stimulation of collagen production by fibroblasts in cell culture. Determine whether neutralization of the expression of integrins on T cells with siRNA or of the integrin function with neu- tralizing antibodies abrogate their regulatory phenotype and the profibrotic effect in co-cultures. Determine whether overexpression of 1V-containing integrins in pulmonary T cells derived from healthy volunteers causes changes toward regulatory profibrotic phenotype. Investigate the ability of CCL18 to directly modulate the phe- notype of normal human pulmonary T cells toward profibrotic, by stimulating the expression, activation, and cell surface binding of TGF-2, and expression of 1V-containing and 22-containing integrins. PUBLIC HEALTH RELEVANCE: Relevance of the Proposed Research to Veterans Health. Excessive scarring, or fibrosis, plays a pivotal role in various diseases of the lungs, liver, and kidneys, in wound granulation, arteriosclerosis and in chronic inflammation in all organs. New knowledge about mechanisms of fibrosis in one organ or disease may be broadly applicable to various diseases in which unwanted excessive scarring occurs. Scarring mechanisms in different diseases may share certain key pathologic components. For example, scarring of the lungs occurs in such diseases as scleroderma, rheumatoid arthritis, hypersensitivity pneumonitis, sarcoidosis, and idiopathic pulmonary fibrosis. Rates of lung fibrosis-associated deaths are even higher in the elderly and male populations, and those subjected to military exposures. These groups are abundantly represented in the veteran population. This proposal is broadly applicable to pulmonary medicine, inflammatory and immune abnormalities, autoimmunity, and immune-mediated fibrosis. Therefore, this research is applicable to the VA patient care mission, the designated priority areas of pulmonary disorders, autoimmune disorders, and wound healing and to the designated research area of chronic diseases.

描述（由申请人提供）： 肺纤维化是慢性炎症过程和环境暴露的常见后果。现有的抗纤维化疗法效率低下。数据表明，T 细胞通常存在于与纤维化相关的患者肺部，并表达整合素，这些整合素可能是定义 T 细胞促纤维化或抗纤维化表型的主开关。本研究的总体目标是阐明表达整合素的 T 细胞在肺部促纤维化和抗纤维化调节中的机制作用。本研究的具体假设是，肺 T 细胞上整合素 1V23、1V25、22 和 1E 的表达和功能活性定义了它们的调节表型（TGF-2+ 类似于 Th3 和 Tregs）通过膜结合的 TGF-2 发挥作用，在肺中持久存在，并刺激胶原蛋白的产生。我们进一步假设，不表达整合素的 T 细胞的肺部浸润导致活性 TGF-2 减少，TNF-1 和 IFN-3 表达升高，随后导致胶原蛋白产生减弱。将解决以下具体目标。 1. 确定肺 T 细胞上整合素表达升高是否与 T 细胞的 TGF-2 相关调节表型和较低的肺功能相关，而不表达整合素的 T 淋巴细胞浸润与炎症表型和较高的肺功能相关。间质性肺疾病患者的功能。 2. 确定体内 CCL18 过表达模型中肺 T 淋巴细胞上整合素表达水平和功能活性的升高是否定义了 a）它们在肺部的长期持续存在，b）整合素的产生、激活和细胞表面结合的增加TGF-2，和c) 胶原蛋白积累。确定 CCL18 过表达对博来霉素诱导的胶原蛋白积累的部分中和是否依赖于肺 T 淋巴细胞上含 1V 整联蛋白表达的减少、活性 TGF-2 表达的减少以及 TNF-1 和 IFN-α 的升高。 3.确定是否可以通过治疗性调节这些整合素介导的促纤维化和抗纤维化机制来调节实验动物肺部的胶原蛋白积累。 3. 确定含有 1V、22 或 1E 的整联蛋白是否促进体外肺 T 细胞和原代成纤维细胞之间的细胞间相互作用、TGF-2 产生和/或激活的增加、TGF-β 的细胞表面结合。 2 通过 T 细胞，并通过细胞培养物中的成纤维细胞刺激胶原蛋白的产生。确定用 siRNA 中和 T 细胞上整合素的表达或用中和抗体中和整合素功能是否会消除共培养物中的调节表型和促纤维化作用。确定来自健康志愿者的肺 T 细胞中含有 1V 的整合素的过度表达是否会导致调节性促纤维化表型的变化。通过刺激 TGF-2 的表达、激活和细胞表面结合以及含 1V 和 22 整联蛋白的表达，研究 CCL18 直接调节正常人肺 T 细胞表型向促纤维化的能力。 公共卫生相关性： 拟议研究与退伍军人健康的相关性。过度的疤痕或纤维化在肺、肝、肾的各种疾病、伤口肉芽形成、动脉硬化和所有器官的慢性炎症中起着关键作用。关于一种器官或疾病的纤维化机制的新知识可能广泛适用于发生不必要的过度疤痕的各种疾病。不同疾病的疤痕机制可能具有某些共同的关键病理成分。例如，肺部疤痕发生在硬皮病、类风湿性关节炎、过敏性肺炎、结节病和特发性肺纤维化等疾病中。老年人、男性以及那些经历过军事暴露的人中，与肺纤维化相关的死亡率甚至更高。这些群体在退伍军人群体中占有相当大的比例。该提案广泛适用于肺科、炎症和免疫异常、自身免疫和免疫介导的纤维化。因此，本研究适用于 VA 患者护理任务、肺部疾病、自身免疫性疾病和伤口愈合的指定优先领域以及慢性病的指定研究领域。