Regulation of Type 1 Diabetes Using Ribozymes

使用核酶调节 1 型糖尿病

• 批准号: 6399867
• 负责人: CHIH-PIN LIU
• 金额: $ 17.5万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-30 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6399867
• 关键词: NOD mouse; Retroviridae; SCID mouse; T lymphocyte; adeno associated virus group; diabetes mellitus therapy; disease /disorder prevention /control; gene therapy; hematopoietic stem cells; insulin dependent diabetes mellitus; nonhuman therapy evaluation; ribozymes; technology /technique development; transfection /expression vector

The overall purpose of this proposal is to develop novel therapeutic approaches to prevent type 1 diabetes by utilizing catalytic RNAs or ribozymes. Type 1 diabetes is a T cell mediated autoimmune disease. The regulatory mechanisms that underlie the development and activation of pathogenic T cells in diabetes are not clear. Previous studies have shown that a breakdown of tolerance and an imbalance of Th1 vs. Th2 type of cytokines with a skewing toward the development of Th1 cells may be involved in the induction and development of type 1 diabetes. It is hypothesized that therapeutic treatment of the disease may be achieved using ribozymes to black the development of diabetogenic Th1 cells or to black adhesion molecules to induce peripheral tolerance of T cells. Ribozymes are enzymatic RNAs that can be engineered to site-specifically cleave targeted RNA and have been effectively used in cell culture and in animal models. For Specific Aim 1, hammerhead ribozymes will first be developed and the most effective ribozymes will then be expressed in T-cells and bone marrow stem cells. Functional expression of the ribozymes will be assayed in vito. Effective combinations of ribozymes and expression systems will be further tested by incorporation into murine retroviral or adeno associated viral (AAV) vectors for delivery into T-cells, bone marrow stem cells and pancreatic islets. In vivo testing of these genetically altered cells in Specific Aim 2 will be carried out in non-obese diabetic (NOB) mice. These studies will critically examine whether or not down regulation of the proposed targets by ribozymes has potential for treating type 1 diabetes.

该提案的总体目的是开发新的治疗方法，通过利用催化RNA或核酶来预防1型糖尿病。 1型糖尿病是一种T细胞介导的自身免疫性疾病。糖尿病中致病性T细胞的发展和激活的调控机制尚不清楚。 先前的研究表明，耐受性的破坏和Th1与Th2型细胞因子的失衡，并向Th1细胞的发展倾斜，可能参与1型糖尿病的诱导和发展。 据推测，该疾病的治疗性治疗可以通过使用核酶使致糖尿病性Th1细胞的发育变黑或使粘附分子变黑以诱导T细胞的外周耐受来实现。 核酶是酶促RNA，其可以被工程化以位点特异性地切割靶向RNA，并且已经有效地用于细胞培养和动物模型中。 对于特异性目标1，将首先开发锤头状核酶，然后在T细胞和骨髓干细胞中表达最有效的核酶。 将在体外测定核酶的功能性表达。 核酶和表达系统的有效组合将通过掺入鼠逆转录病毒或腺相关病毒（AAV）载体中以递送到T细胞、骨髓干细胞和胰岛中来进一步测试。 将在非肥胖糖尿病（NOB）小鼠中对特定目标2中的这些遗传改变细胞进行体内试验。 这些研究将严格检查核酶下调所提出的靶点是否具有治疗1型糖尿病的潜力。