Beta-Globin Gene Correction Using Peptide Nucleic Acids for the Treatment of Sick

使用肽核酸校正β-珠蛋白基因来治疗疾病

基本信息

批准号：
7997382
负责人：
Gerald Francis Vovis
金额：
$ 22.85万
依托单位：
HELIX THERAPEUTICS, LLC
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-08-01 至 2011-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7997382
关键词：
Address Affect Allogenic Amino Acids Autologous Autologous Transplantation Blood CD34 gene Cell Survival Cells Clinical Research Cost of Illness DNA DNA Repair Disease Engraftment Erythrocytes FDA approved Frequencies Gene Targeting Gene Transduction Agent Genes Genetic Genetic Recombination Globin Hematopoietic Stem Cell Transplantation Hematopoietic stem cells Hemoglobin Hereditary Disease Human Human Genetics In Situ Individual Inherited Laboratory Research Life Expectancy Mammalian Cell Measures Methods Modality Modification Morbidity - disease rate Mutation Oligonucleotides Oxygen Patients Peptide Nucleic Acids Phase Point Mutation Preventive Procedures Production Proteins Sickle Cell Anemia Site Stem cell transplant Stem cells Techniques Technology Testing Therapeutic Transplantation United States Viral Viral Vector Work beta Globin gene correction gene repair high risk human disease hydroxyurea immunodeficient mouse model interest minimally invasive mortality mouse model public health relevance research clinical testing research study success therapeutic gene triple helix tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): This is a proposal to test the feasibility of using triplex-forming oligonucleotides to correct the sickle cell disease (SCD) mutation in human CD34+ cells. SCD is one of the most common human genetic diseases and is the result of one specific inherited mutation in the 2-globin gene. Hydroxyurea is currently the only FDA-approved treatment for SCD. Both supportive and preventive measures remain the mainstay of treatment for this disease. Allogeneic hematopoietic stem cell transplantation is curative, although such a treatment typically requires an HLA-matched donor, is associated with significant morbidity and is very expensive. However, genetic modification of autologous CD34+ cells is becoming a viable therapeutic modality. Recent studies suggest that this technique can be used to treat hemoglobin disorders. Current methods for correcting mutations in CD34+ cells require non-directed insertion of viral vectors. This method is expensive, has low success rates and carries a high risk of induced tumorigenesis. Some non-viral methods have been developed for directly correcting the genes of interest in CD34+ cells. However, these current approaches are expensive and have a low rate of success. To address the above weaknesses in current curative methods, we propose to test the feasibility of using our triplex-forming oligonucleotide approach. Our procedure has been shown to stimulate recombination in mammalian cells by the ability of triple helices to provoke DNA repair and, thus, sensitize the target site to recombination. This technology constitutes minimally invasive gene repair, as gene modification occurs in situ via use of the cell's own DNA repair machinery, without the need for viral vectors. Helix Therapeutics was formed to commercialize this technology for treating common human diseases, including SCD. Our approach promises to be safe and inexpensive. In the proposed studies, we will determine an effective combination of triplex and donor DNA molecules, as well as an effective cellular delivery method for our gene targeting molecules. Our primary proof of principle will be to demonstrate that this method can induce directed mutations of the 2-globin gene at a rate of 10% or higher, which is the expected level required to become clinically effective. We will carry out this proof of principle test in human CD34+ cells and evaluate whether the cells are able to engraft and properly differentiate in a mouse model of stem cell transplantation. These experiments will constitute a proof-of-concept study. Additional improvements to the methods and other studies necessary for filing an IND for clinical testing will take place in a Phase II application. PUBLIC HEALTH RELEVANCE: Sickle cell disease (SCD), also called sickle cell anemia, is among the most common human genetic disorders and is the result of one specific inherited mutation in the 2-globin gene, which is involved in the synthesis of hemoglobin. Hemoglobin is the protein in red blood cells that carries oxygen. Helix Therapeutics is proposing to develop a therapeutic gene targeting agent to correct, in human hematopoietic stem cells, the mutation responsible for causing SCD and, thus, cure the disease permanently.

描述（由申请人提供）：这是一项测试使用三链体形成寡核苷酸来纠正人类 CD34+ 细胞中镰状细胞病（SCD）突变的可行性的提案。 SCD 是最常见的人类遗传疾病之一，是 2-珠蛋白基因中一种特定遗传突变的结果。羟基脲是目前 FDA 唯一批准的治疗 SCD 的药物。支持性和预防性措施仍然是治疗这种疾病的主要方法。同种异体造血干细胞移植具有治愈性，但这种治疗通常需要 HLA 匹配的供体，且与显着的发病率相关并且非常昂贵。然而，自体 CD34+ 细胞的基因修饰正在成为一种可行的治疗方式。最近的研究表明，这种技术可用于治疗血红蛋白疾病。目前纠正 CD34+ 细胞突变的方法需要非定向插入病毒载体。该方法成本高、成功率低、诱发肿瘤的风险高。一些非病毒方法已被开发用于直接校正 CD34+ 细胞中感兴趣的基因。然而，目前的这些方法成本高昂且成功率较低。为了解决当前治疗方法中的上述弱点，我们建议测试使用我们的三链体形成寡核苷酸方法的可行性。我们的程序已被证明可以通过三螺旋激发 DNA 修复的能力来刺激哺乳动物细胞中的重组，从而使靶位点对重组敏感。这项技术构成了微创基因修复，因为基因修饰是通过使用细胞自身的 DNA 修复机制在原位发生的，而不需要病毒载体。 Helix Therapeutics 的成立是为了将这项技术商业化，用于治疗常见的人类疾病，包括 SCD。我们的方法有望安全且廉价。在拟议的研究中，我们将确定三链体和供体 DNA 分子的有效组合，以及基因靶向分子的有效细胞递送方法。我们的主要原理证明将证明该方法可以以 10% 或更高的速率诱导 2-珠蛋白基因的定向突变，这是临床有效所需的预期水平。我们将在人类 CD34+ 细胞中进行这一原理验证测试，并评估这些细胞是否能够在干细胞移植的小鼠模型中移植并正确分化。这些实验将构成概念验证研究。对提交临床试验 IND 所需的方法和其他研究的进一步改进将在 II 期申请中进行。公共卫生相关性：镰状细胞病 (SCD) 也称为镰状细胞性贫血，是最常见的人类遗传性疾病之一，是 2-珠蛋白基因（参与血红蛋白合成）的一种特定遗传突变的结果。血红蛋白是红细胞中携带氧气的蛋白质。 Helix Therapeutics 提议开发一种治疗性基因靶向剂，以纠正人类造血干细胞中导致 SCD 的突变，从而永久治愈该疾病。