COORDINATE REGULATION OF APOPTOSIS AND CELL CYCLE IN RA

RA中细胞凋亡和细胞周期的协调调控

• 批准号: 6789963
• 负责人: Harris R Perlman
• 金额: $ 11.67万
• 依托单位: SAINT LOUIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-15 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6789963
• 关键词: BCL2 gene /protein; CD95 molecule; apoptosis; cell cycle; cell proliferation; cellular pathology; fibroblasts; gene therapy; human tissue; laboratory rat; membrane permeability; mitochondrial membrane; nonhuman therapy evaluation; rheumatoid arthritis; ribozymes; synovial membrane

The work proposed for this grant focuses on the coordinate regulation of proliferation and apoptosis in rheumatoid arthritis (RA). Analysis of human RA synovial tissues (ST) sections revealed increased rates of synovial fibroblast proliferation and low rates of apoptosis, though the functional significance of reduced apoptosis remains to be elucidated. However, analysis of the rates of in vivo proliferation and apoptosis are limited in human-STs as tissue sections were taken late in disease course. Thus, utilization of animal models is vital for an understanding of the molecular pathways of proliferation and apoptosis in RA. Recently, adenoviral mediated delivery of Fas ligand (Ad-FasL), a known apoptotic inducer ameliorated experimental arthritis, suggesting that enhancing the rate of apoptosis by gene therapy may be a potential effective therapy. A caveat to Ad-FasL therapy is that high levels of Fas ligand is cytotoxic to many tissues of the body, thus development of other genes to be delivered to the RA joint is essential. We demonstrated that the anti-apoptotic protein and cell cycle modulator, Bcl-2 was highly expressed in RA compared with osteoarthritis synovial tissues, particularly in the CD68- negative, fibroblast-like synoviocyte population. In order to determine the importance of endogenous Bcl-2, an adenoviral vector expressing a hammerhead ribozyme to Bcl-2 (Ad-Rbz-Bcl-2) mRNA was employed. Ad-Rbz-Bcl-2 infection resulted in reduced Bcl-2 expression and cell viability in synovial fibroblasts isolated from RA-synovial tissues. In addition, Ad-Rbz-Bcl-2- induced mitochondrial permeability transition, cytochrome c release, activation of caspases 9 and 3, and DNA fragmentation. These data suggest that Bcl-2 is necessary for synovial fibroblast survival. In this proposal we describe studies to delineate the mechanism of the induction of mitochondrial permeabilty transition following Ad-Rbz-Bcl-2 infection. In addition we will investigate whether adenoviral mediated delivery of the Bcl-2 ribozyme is efficient in ameliorating adjuvant- induced arthritis in rats. The expected outcome is the suppression of AIA through the inhibition of fibroblast proliferation and increased apoptosis. This approach could lead to the development of a new therapeutic strategy for gene therapy in patients with rheumatoid arthritis.

这项资助的工作重点是类风湿关节炎（RA）中细胞增殖和细胞凋亡的协调调节。 对人RA滑膜组织（ST）切片的分析显示滑膜成纤维细胞增殖率增加，细胞凋亡率降低，但细胞凋亡减少的功能意义仍有待阐明。 然而，由于组织切片是在疾病过程的后期进行的，因此对人ST的体内增殖和凋亡率的分析是有限的。因此，利用动物模型是至关重要的，了解在RA增殖和凋亡的分子途径。最近，腺病毒介导的Fas配体（Ad-FasL），一种已知的凋亡诱导剂的交付改善实验性关节炎，这表明，通过基因治疗提高细胞凋亡率可能是一个潜在的有效治疗。 Ad-FasL治疗的警告是高水平的Fas配体对身体的许多组织具有细胞毒性，因此开发其他基因以递送至RA关节是必要的。我们证明了抗凋亡蛋白和细胞周期调节剂Bcl-2在RA中的表达高于骨关节炎滑膜组织，特别是在CD 68阴性的成纤维细胞样滑膜细胞群体中。 为了确定内源性Bcl-2的重要性，使用表达针对Bcl-2的锤头状核酶（Ad-Rbz-Bcl-2）mRNA的腺病毒载体。 Ad-Rbz-Bcl-2感染导致从RA滑膜组织分离的滑膜成纤维细胞中Bcl-2表达和细胞活力降低。 此外，Ad-Rbz-Bcl-2诱导线粒体通透性转换，细胞色素c释放，半胱氨酸天冬氨酸蛋白酶9和3的激活，以及DNA片段化。这些数据表明，Bcl-2是滑膜成纤维细胞存活所必需的。 在这个建议中，我们描述的研究，描绘的机制，诱导线粒体通透性转换后的Ad-Rbz-Bcl-2感染。 此外，我们将研究腺病毒介导的Bcl-2核酶递送是否有效改善大鼠佐剂诱导的关节炎。 预期的结果是通过抑制成纤维细胞增殖和增加细胞凋亡来抑制AIA。 这种方法可能会导致开发一种新的治疗策略，用于类风湿性关节炎患者的基因治疗。

项目成果

BH3-only proteins in rheumatoid arthritis: potential targets for therapeutic intervention.

• DOI: 10.1038/onc.2009.54
• 发表时间: 2008-12
• 期刊: Oncogene
• 影响因子: 8
• 作者: Hutcheson J;Perlman H
• 通讯作者: Perlman H

Regulation of apoptosis and cell cycle activity in rheumatoid arthritis.

类风湿关节炎中细胞凋亡和细胞周期活性的调节。

• DOI: 10.2174/1566524013363429
• 发表时间: 2001
• 期刊: Current molecular medicine
• 影响因子: 2.5
• 作者: Perlman,H;Pagliari,LJ;Volin,MV
• 通讯作者: Volin,MV

A cell-cycle independent role for p21 in regulating synovial fibroblast migration in rheumatoid arthritis.

p21在调节类风湿关节炎中的滑膜成纤维细胞迁移中的独立作用。

• DOI: 10.1186/ar1999
• 发表时间: 2006
• 期刊: ARTHRITIS RESEARCH & THERAPY
• 影响因子: 4.9
• 作者: Woods, James M;Klosowska, Karolina;Spoden, Darrin J;Stumbo, Nataliya G;Paige, Douglas J;Scatizzi, John C;Volin, Michael V;Rao, Malathi S;Perlman, Harris
• 通讯作者: Perlman, Harris