Development of a Novel Method for Inhibiting Atherosclerosis in Diabetes

开发抑制糖尿病动脉粥样硬化的新方法

• 批准号: 7109891
• 负责人: DAVID Robert CLEMMONS
• 金额: $ 21.03万
• 依托单位: VASCULAR PHARMACEUTICALS
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7109891
• 关键词: atherosclerosis; biological signal transduction; cardiovascular disorder prevention; cell migration; cell proliferation; cysteine; diabetes mellitus therapy; growth factor receptors; immunologic substance development /preparation; immunotherapy; insulinlike growth factor; integrins; laboratory mouse; monoclonal antibody; polymerase chain reaction; protein structure function; smooth muscle; swine; therapy design /development

DESCRIPTION (provided by applicant): The long term objective of the proposed studies is to develop a monoclonal antibody that can be administered safely to patients with diabetes to determine if it can inhibit atherosclerosis. Insulin-like growth factor-l (IGF-I) has been implicated in initiating the proliferative phase of atherosclerotic lesion development but inhibiting IGF-I receptor activity may lead to toxicity. IGF-I functions cooperatively with the aV?3 integrin to stimulate smooth muscle celll(SMC) division and migration which are related to the development of atherosclerosis. Therefore the purpose of these studies will be to determine if targeting the aV?3 integrin disrupts IGF-I stimulated atherosclerotic lesion progression. A specific monoclonal antibody directed against the cysteine loop (amino acids 177-183) of the 03 subunit of the aV|33 integrin will be prepared and tested to determine if it inhibits IGF-I stimulated SMC growth and migration in vitro and if this response is due disruption of IGF-I receptor linked signaling mechanisms that lead to stimulation of cell growth. To determine if this antibody has efficacy in vivo Fab fragments of the antibody will be prepared and will be infused into the arteries of pigs that are developing atherosclerotic lesions. A control antibody will be infused into one of the lesion areas to rule out the possibility of nonspecific effects due to autoimmunization. To obtain the structural information that will be necessary to prepare a humanized form of the antibody the clone of cells secreting the antibody will be isolated and the cDNAs corresponding to the heavy and light chains will be amplified using RTPCR. The cDNAs will be isolated, cloned and their sequences determined. The antibody structure will be used to design humanized form of the antibody in phase II. These studies should definitively test the hypothesis that inhibiting IGF-I actions by inhibiting ligand occupancy of aV?33 in blood vessel walls inhibits atherosclerotic lesion progression. If these results show that this approach is feasible a humanized version of this monoclonal would be prepared, purified and tested for toxicity. The long term objective would be to administer this antibody to diabetic patients with atherosclerosis who have wide spread diffuse disease that is not amenable to current therapies such as drug eluting stents. Since atherosclerosis is the cause of death in 80% of diabetics and there is no currently available therapy that specifically treats atherosclerosis in diabetics there is a major need for new therapies that are directed toward the treatment of this complication.

描述（由申请人提供）：拟议研究的长期目标是开发一种单克隆抗体，可以安全地给药给糖尿病患者，以确定它是否可以抑制动脉粥样硬化。胰岛素样生长因子- 1 （IGF-I）参与了动脉粥样硬化病变发展的增殖阶段，但抑制IGF-I受体活性可能导致毒性。IGF-I与aV？3整合素刺激平滑肌细胞（SMC）的分裂和迁移，这与动脉粥样硬化的发展有关。因此，这些研究的目的将是确定是否靶向aV？整合素破坏igf - 1刺激的动脉粥样硬化病变进展。一种针对aV bbbb33整合素03亚基的半胱氨酸环（氨基酸177-183）的特异性单克隆抗体将被制备和测试，以确定它是否抑制IGF-I刺激的SMC生长和体外迁移，以及这种反应是否由于IGF-I受体相关信号机制的破坏而导致刺激细胞生长。为了确定这种抗体在体内是否有效，将制备抗体的Fab片段并将其注入正在发生动脉粥样硬化病变的猪的动脉中。对照抗体将被注入一个病变区域，以排除由于自身免疫而产生非特异性影响的可能性。为了获得制备人源化抗体所需的结构信息，将分离分泌抗体的细胞克隆，并使用RTPCR扩增重链和轻链对应的cdna。这些dna将被分离、克隆并确定其序列。该抗体结构将用于II期抗体的人源化设计。这些研究应该明确地验证通过抑制配体占用aV来抑制IGF-I作用的假设。33在血管壁抑制动脉粥样硬化病变进展。如果这些结果表明这种方法是可行的，将制备、纯化和测试该单克隆的人源化版本的毒性。长期目标是将这种抗体用于动脉粥样硬化的糖尿病患者，这些患者有广泛的弥漫性疾病，不适合目前的治疗方法，如药物洗脱支架。由于动脉粥样硬化是80%的糖尿病患者死亡的原因，目前还没有专门治疗糖尿病患者动脉粥样硬化的有效疗法，因此迫切需要针对这一并发症的新疗法。