IL-1 RECEPTOR ANTAGONIST GENE THERAPY FOR THE TREATMENT OF INFLAMMATORY DISEASES

我们的实验室已经证明，用重组IL-1治疗 受体拮抗剂（IL-1 ra）通过重复注射显著抑制 炎症和组织损伤。 另外我们 最近发现溃疡性结肠炎和克罗恩病患者 与正常人相比， 对照组和自限性感染性结肠炎，这表明患者 患有炎症性肠病的患者可能缺乏 产生IL-1 ra。 因此，恢复生理平衡 IL-1 ra和IL-1之间的关系可能有助于减少肠道 炎症 我们的假设是， 可稳定生产和递送重组体的移植系统 IL-1 ra进入体循环可能代表了一个重要的进展 我们治疗炎症性疾病的能力， 肠道疾病和肝脏疾病，其中IL-1已被证明发挥作用， 一个重要的角色 这个假设将通过完成 具体目的如下：1）构建质粒载体， 兔IL-1 ra的分泌或细胞内形式 cDNA，并产生稳定转染的肌源性细胞系或原代 培养的成肌细胞合成大量重组IL-1 ra， 体外; 2）肌内注射培养的成肌细胞， 邻近正常的肌肉纤维，并将提供大量的分泌 体内IL-1 ra。 这些研究将采用最先进的分子 生物学技术结合经验证的免疫测定方法学。 的 该提案的最终目标是应用从这些 治疗和预防肝脏和胃肠道炎症的研究 通过利用基因增强IL-1 ra表达的疾病 治疗方法

Our laboratory has demonstrated that treatment with recombinant IL-1 receptor antagonist (IL-1ra) by repeated injections markedly suppresses inflammation and tissue damage in experimental colitis. In addition, we have recently shown that patients with ulcerative colitis and Crohn's disease have a decreased intestinal IL-1ra/IL-1 ratio compared to normal controls and self-limited infectious colitis, suggesting that patients with inflammatory bowel disease may have a deficit in their ability to produce IL-1ra. Therefore, restoration of the physiological balance between IL-1ra and IL-1 may prove beneficial in reducing intestinal inflammation. Our hypothesis is that development of a cellular transplantation system that can stably produce and deliver recombinant IL-1ra into the systemic circulation may represent an important advance in our ability to treat inflammatory disease, including inflammatory bowel disease and liver diseases in which IL-1 have been shown to play an important role. This hypothesis will be investigated by accomplishing the following specific aims: 1) construct plasmid vectors containing either the secreted(s) or the intracellular (ic) form of rabbit IL-1ra cDNA, and produce stably transfected myogenic cell lines or primary cultured myoblasts synthesizing large amounts of recombinant IL-1ra in vitro; 2) inject intramuscularly, cultured myoblasts that will fuse into adjacent normal muscle fibers and will deliver large amounts of secreted IL-1ra in vivo. These studies will employ state-of-the art molecular biology techniques combined with validated immunoassay methodology. The ultimate goal of this proposal is to apply information derived from these studies to treat and prevent liver and gastrointestinal inflammatory diseases through augmentation of IL-1ra expression by utilizing gene therapy methodologies.