PHARMACOLOGICAL & GENE THERAPY OF ARPKD--FROM CELL TO ANIMAL

药理

• 批准号: 6493083
• 负责人: Ellis David Avner
• 金额: $ 16.2万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2002-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6493083
• 关键词: antisense nucleic acid; autosomal recessive trait; cell line; combination therapy; disease /disorder model; enzyme inhibitors; epidermal growth factor; gene therapy; growth factor receptors; kidney pharmacology; laboratory mouse; liposomes; polycystic kidney; protein tyrosine kinase; receptor mediated endocytosis; transfection; transfection /expression vector; transforming growth factors

Description: (Taken directly from the application) The objective of this research is to develop and test therapies for autosomal recessive polycystic kidney disease (ARPKD), which are specifically designed to target key processes in disease pathogenesis. The overall hypothesis to be tested is that interruption or therapeutic targeting of abnormalities in the epidermal growth factor (EGF)-transforming growth factor alpha (TGF-alpha) epidermal growth factor receptor (EGFR) axis in ARPKD will result in structural and functional improvement in cystic kidney disease. Published data from a number of laboratories demonstrate that altered EGFR expression in cystic tubular epithelial cells is common to human and murine ARPKD as well as autosomal dominant polycystic kidney disease (ADPKD). Although the specific projects proposed will focus on animal models of ARPKD and human ARPKD cells, therapies developed during the course of this study may be generalizable to ADPKD as well. Studies will be conducted in primary and immortalized cell lines, organ culture and intact animals. Pharmacologic, genetic and cell biology techniques will be employed to accomplish the following Specific Aims: 1. Develop therapeutic strategies aimed at interrupting EGF-TGFalpha-EGFR axis activity in ARPKD. Specific hypotheses to be tested are: a.) Inhibition of EGFR activity results in sustained improvement of renal function without toxicity in ARPKD; b.) Inhibition of the production and/or processing of biologically active ligands (EGF/TGFalpha ) has therapeutic effectiveness in ARPKD: and c.) Combined inhibition of EGFR activity and processing/production of EGF/TGFalpha is more effective and less toxic than either therapy given alone. Methods to be utilized in the proposed studies include in vitro/vivo trials of EGFR tyrosine kinase inhibitors, agents which alter ligand production, and antisense oligonucleotides. 2. Develop new strategies for gene therapy in ARPKD. Specific hypotheses to be tested are: a.) Gene replacement following disease development can alter disease progression in ARPKD; b.) Effectiveness and specificity of gene therapy can be improved by utilizing elements of the EGF/TGFalpha /EGFR axis to augment gene delivery; and c.) Genetic therapy directed at targeting key pathogenic processes following disease development can alter disease progression in ARPKD. Methods to be utilized in the proposed studies include liposome mediated gene transfer, adeno-associated virus-mediated gene transfer, and the use of EGFR-ligand or antibody to deliver genes by receptor mediated endocytosis. These studies will provide new scientific insights and information which may be useful in the design of specific treatments for human ARPKD and ADPKD.

描述：（直接摘自申请）这项研究的目的是开发和测试常染色体隐性多囊肾病 (ARPKD) 的治疗方法，这些治疗方法专门针对疾病发病机制的关键过程而设计。待测试的总体假设是，针对 ARPKD 中表皮生长因子 (EGF) 转化生长因子 α (TGF-α) 表皮生长因子受体 (EGFR) 轴异常的中断或治疗目标将导致 ARPKD 的结构和功能改善。囊性肾病。许多实验室公布的数据表明，囊性肾小管上皮细胞中 EGFR 表达的改变在人类和小鼠 ARPKD 以及常染色体显性多囊肾病 (ADPKD) 中很常见。尽管提出的具体项目将重点关注 ARPKD 动物模型和人类 ARPKD 细胞，但本研究过程中开发的疗法也可能适用于 ADPKD。研究将在原代细胞系和永生化细胞系、器官培养物和完整动物中进行。将采用药理学、遗传和细胞生物学技术来实现以下具体目标： 1. 制定旨在中断 ARPKD 中 EGF-TGFα-EGFR 轴活性的治疗策略。要测试的具体假设是： a.) 抑制 EGFR 活性可导致肾功能持续改善，且对 ARPKD 没有毒性； b.) 抑制生物活性配体 (EGF/TGFα) 的产生和/或加工对 ARPKD 具有治疗效果：和 c.) 联合抑制 EGFR 活性和 EGF/TGFα 的加工/产生比其他药物更有效且毒性更小单独给予任一疗法。拟议研究中使用的方法包括 EGFR 酪氨酸激酶抑制剂、改变配体产生的药物和反义寡核苷酸的体外/体内试验。 2. 制定 ARPKD 基因治疗新策略。要测试的具体假设是： a.) 疾病发展后的基因替换可以改变 ARPKD 的疾病进展； b.) 通过利用 EGF/TGFα/EGFR 轴的元件来增强基因递送，可以提高基因治疗的有效性和特异性； c.) 针对疾病发展后关键致病过程的基因治疗可以改变 ARPKD 的疾病进展。拟议研究中使用的方法包括脂质体介导的基因转移、腺相关病毒介导的基因转移以及使用EGFR配体或抗体通过受体介导的内吞作用递送基因。这些研究将提供新的科学见解和信息，可能有助于设计人类 ARPKD 和 ADPKD 的具体治疗方法。