Antisense Treatment for Thalassemia. Preclinical study.

地中海贫血的反义治疗。

• 批准号: 7097163
• 负责人: RYSZARD KOLE
• 金额: $ 58.86万
• 依托单位: ERCOLE BIOTECH, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-30 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7097163
• 关键词: RNA splicing; antisense nucleic acid; chemical conjugate; clinical research; erythroid stem cell; hemoglobin; human subject; human tissue; laboratory mouse; oligonucleotides; polymerase chain reaction; precursor mRNA; spliceosomes; thalassemia; therapy design /development

DESCRIPTION (provided by applicant): The ultimate goal of this project is treatment of beta-thalassemia, a genetic blood disease, with antisense oligonucleotides. The project is based on the technology developed by the PI in which antisense oligonucleotides are used to restore correct functioning of a defective gene rather than, as commonly practiced, to inhibit the expression of an undesirable gene. Specifically, oligonucleotides are used to reverse aberrant splicing of human beta-globin pre-mRNA by blocking splice sites activated by a mutation IVS2-654 in intron 2 of the beta-globin gene. Aberrant splicing is also reversed for betaE mutation in exon 1. The oligonucleotides not only inhibit aberrant splicing but, by forcing the spliceosomes to form on the existing correct splice sites, restore correct splicing of beta-globin pre-mRNA, which is translated into the beta-globin polypeptide, ultimately restoring expression of hemoglobin. The main goal of this Phase I proposal is to improve the effectiveness of morpholino oligonucleotides targted to the aberrant IVS2-654 splice site by conjugating them with penetrating tat-like peptides. The proposed work will be carried mostly in erythroid cells from the IVS2-654 mice, in vivo in this mouse model of thalassemia and in cells from thalassemia patients with IVS2-654 thalassemia. The Specific Aims of the project are: 1) To test if morpholino-peptide conjugates effectively correct splicing of beta-globin pre-mRNA and restore expression of hemoglobin in vitro in erythroid cells from IVS2-654 mouse. 2) To test if morpholino-peptide conjugates effectively correct splicing of beta-globin pre-mRNA and restore expression of hemoglobin in vivo in IVS2-654 mouse. 3) To confirm that the most effective oligonucleotides identified in Specific Aims 1 and 2 are effective in erythroid cells from patients with thalassemia.

描述（由申请人提供）：该项目的最终目的是治疗β-甲性疾病（一种遗传性血液疾病），具有反义寡核苷酸。该项目基于PI开发的技术，在该技术中，反义寡核苷酸用于恢复有缺陷基因的正确功能，而不是通常实用的，以抑制不良基因的表达。具体而言，寡核苷酸用于通过阻断β-珠蛋白基因内含子2中突变IVS2-654激活的剪接位点来扭转人β-珠蛋白前MRNA的异常剪接。 Aberrant splicing is also reversed for betaE mutation in exon 1. The oligonucleotides not only inhibit aberrant splicing but, by forcing the spliceosomes to form on the existing correct splice sites, restore correct splicing of beta-globin pre-mRNA, which is translated into the beta-globin polypeptide, ultimately restoring expression of hemoglobin.该阶段I提案的主要目标是通过将它们与穿透性Tat类肽结合到异常的IVS2-654剪接位点上的形态寡核苷酸的有效性。所提出的工作将主要用于来自IVS2-654小鼠的红细胞细胞，在这种小鼠thalassymia小鼠模型中，以及来自IVS2-654 Thalassya的Thalassybia患者的细胞中。该项目的具体目的是：1）测试吗啡肽是否有效地纠正β-珠蛋白pre-mRNA的剪接并从IVS2-654小鼠的红细胞细胞中体外恢复体外的血红蛋白表达。 2）测试吗啡 - 肽是否有效地纠正β-珠蛋白前MRNA的剪接并恢复体内IVS2-654小鼠体内血红蛋白的表达。 3）确认在特定目的1和2中鉴定出的最有效的寡核苷酸在丘脑贫血患者的红细胞细胞中有效。