Clinical Application of Cell Cycle Control Therapy to Rheumatoid Arthritis

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>对治疗类风湿性关节炎（RA）动物模型有效。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抑制依赖性和CDK抑制非依赖性途径抑制炎症介质和蛋白酶的表达。由于腺病毒载体基因治疗中的致命事故，人们对腺病毒载体的安全性提出了更高的要求。通过修饰表达Arg-Gly-Asp（RGD）基序的纤维外壳蛋白或携带部分35型腺病毒纤维分子的5型腺病毒（Ad 5）载体，提高了Ad 5对类风湿性滑膜成纤维细胞（RSF）的基因转移效率。与非- ...更多信息 3）CDKI生物制剂的开发：研究了无病毒载体的CDKI治疗。如果HIV跨膜结构域（达特）被缀合，则蛋白质可以在细胞内递送。TAT偶联的p16^<INK4a>融合蛋白抑制RSF增殖，并观察其对RA动物模型的治疗作用。4）合成小分子CDKI治疗RA：合成的sm-CDKI化合物对RA动物模型滑膜炎有效，我们已申请了两种sm-CDKI化合物用于RA治疗的专利<INK4a><Cip1><Cip1><INK4a>。在<INK4a>其他成纤维细胞中未观察到这种p16 β诱导。利用荧光差异显示或基因芯片技术，我们筛选了一组可能参与p21^<Cip1>-p16^<INK4a>通路的分子。其中一种与怀孕有关的候选分子增加了p16<INK4a>启动子的活性。

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