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Clinical Application of Cell Cycle Control Therapy to Rheumatoid Arthritis

细胞周期调控疗法在类风湿性关节炎中的临床应用

基本信息

批准号：
13854014
负责人：
MIYASAKA Nobuyuki
金额：
$ 78.62万
依托单位：
Tokyo Medical and Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (S)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2005
项目状态：
已结题

项目摘要

Forced expression of a cyclin-dependent kinase inhibitor (CDKI) gene, p16^<INK4a> or p21^<Cip1> in the synovial tissues was effective in treating animal models of rheumatoid arthritis (RA). Subsequently, we have studied molecular mechanism and application of cell cycle control for treatment of rheumatoid arthritis (RA).1) Anti-inflammatory effects of CDKI gene therapy : CDKI gene therapy suppressed expression of inflammatory mediators and proteinases via both CDK inhibition dependent- and CDK inhibition independent-pathways.2) Improvement of gene transfer efficacy by modified adenovirses : Because of a fatal accident by gene therapy using adenovirus vector, safety of adenovirus vector is strongly demanded. Efficacies of gene transfer to rheumatoid synovial fibroblasts (RSF) by type 5 adenoviruses (Ad5) vector were improved by modification fiber-coat proteins expressing Arg-Gly-Asp (RGD) motif or by Ad5 carrying a part of the fiber molecule of adenovirus serotype 35. Comparing with non- … More modified Ad5, fewer quantities of these fiber-modified Ad5 vectors were required to treat synovitis of RA animal model.3) Development of CDKI-'biologics : CDKI therapy without viral vector was studied. Protein can be delivered intracellularly if it HIV transmembrane domain (TAT) is conjugated. TAT-conjugated p16^<INK4a> fusion protein inhibited RSF proliferation, and therapeutic effects for RA animal model is investigated.4) Synthetic small molecule (sm)-CDKI for treatment of RA : Some synthetic sm-CDKI compounds were effective for synovitis of RA animal model, and we have applied patents of two sm-CDKI compounds for RA therapy.5) Selective p16^<INK4a> induction in RSF by p21^<Cip1> : Forced expression of p21^<Cip1> in RSF induced p16^<INK4a> expression. This p16^<INK4a> induction was not observed in the other fibroblasts. Using fluorescent differential display or GeneChip (Affimetrix) techniques, we screened a group of molecules that might be involved in the p21^<Cip1>-p16^<INK4a> pathway. One of the candidate molecules, that is related to pregnancy, increased p16^<INK4a> promoter activity Less

在滑膜组织中强制表达细胞周期蛋白依赖性激酶抑制剂（CDKI）基因p16^<INK4a>或p21^<Cip1>可有效治疗类风湿性关节炎（RA）动物模型。随后，我们研究了细胞周期调控在类风湿关节炎（RA）治疗中的分子机制和应用。1) CDKI基因治疗的抗炎作用：CDKI基因治疗通过CDK抑制依赖通路和CDK抑制独立通路抑制炎症介质和蛋白酶的表达。2)修饰腺病毒提高基因转移效果：由于使用腺病毒载体进行基因治疗的致命事故，对腺病毒载体的安全性提出了强烈要求。5型腺病毒（Ad5）载体通过修饰表达arg - gy - asp （RGD）基序的纤维包被蛋白或Ad5载体携带部分35型腺病毒纤维分子，提高了基因转染类风湿滑膜成纤维细胞（RSF）的效果。与未修饰Ad5相比，这些纤维修饰Ad5载体治疗RA动物滑膜炎所需的数量较少。3) CDKI-’生物制剂的发展：研究了无病毒载体的CDKI治疗。结合HIV跨膜结构域（TAT）的蛋白可以在细胞内传递。tat偶联p16^<INK4a>融合蛋白抑制RSF增殖，并研究其对RA动物模型的治疗作用。4)合成小分子(sm)-CDKI治疗RA：一些合成的sm-CDKI化合物对RA动物模型滑膜炎有效，我们已经申请了两种sm-CDKI化合物治疗RA的专利。5) p21^<Cip1>在RSF中选择性诱导p16^<INK4a>: p21^<Cip1>在RSF诱导p16^<INK4a>表达中强制表达。在其他成纤维细胞中未观察到p16^<INK4a>诱导。利用荧光差分显示或基因芯片（affmetrix）技术，我们筛选了一组可能参与p21^<Cip1>-p16^<INK4a>通路的分子。其中一种候选分子与妊娠有关，可使p16^<INK4a>启动子活性增加较少