Triplex-mediated DNA repair as a novel therapy for Hurler Syndrome patients

三重介导的 DNA 修复作为 Hurler 综合征患者的新型疗法

• 批准号: 8454945
• 负责人: Elizabeth Peterson-Roth
• 金额: $ 26.82万
• 依托单位: L2 DIAGNOSTICS, LLC
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-30 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8454945
• 关键词: Address; Affect; Allogenic; Autologous; Back; Binding; Blood; Blood - brain barrier anatomy; CD34 gene; Caucasians; Caucasoid Race; Cell Therapy; Cells; Clinical; Clinical Research; Collection; DNA; DNA Repair; Deterioration; Development; Diagnostic; Disease; Enzymes; Fabry Disease; Frequencies; Gaucher Disease; Gene Mutation; Gene Targeting; Gene-Modified; Genes; Genetic; Genetic Recombination; Genome; Harvest; Health Personnel; Hematologist; Hematopoietic Stem Cell Transplantation; Hereditary Disease; Hospital Formularies; Hospitals; Human; Immune; Inborn Errors of Metabolism; Incidence; L-Iduronidase; Laboratories; Letters; Live Birth; Lysosomal Storage Diseases; Marketing; Mediating; Modification; Morbidity - disease rate; Mucopolysaccharidosis I H; Multiple Organ Failure; Mus; Mutation; Neuraxis; Neurologic; Nonsense Mutation; Oligonucleotides; Patients; Peptide Nucleic Acids; Pharmacologic Substance; Phase; Physiological; Process; Proteins; Protocols documentation; Qualifying; Recombinant Proteins; Specialist; Stem cells; Supportive care; Symptoms; Technology; Testing; Therapeutic; TimeLine; Transplantation; United States; Vial device; Viral Vector; Work; cost; design; disease-causing mutation; effective therapy; enzyme replacement therapy; genetic element; in vivo; monocyte; mortality; novel; phase 1 study; repaired; success; sugar; therapeutic gene

DESCRIPTION (provided by applicant): This is a proposal to test the feasibility of using triplex molecules for use as a therapeutic to correct Hurler Syndrome (HS). Lysosomal storage diseases (LSDs), such as HS, represent a diverse collection of monogenic disorders that have devastating clinical features. LSDs are caused by mutations in genes that encode lysosomal enzymes and other proteins that are critical for lysosomal function. HS is caused by mutations in the ¿ -L-iduronidase gene (IDUA). For many LSDs, treatment consists of enzyme replacement therapy and supportive care. However, enzyme replacement therapy costs from $90,000 to $565,000 per patient. Alternatively, allogeneic hematopoietic stem cell transplantation (HSCT) represents a highly effective treatment. However, allogeneic HSCT requires an HLA-matched donor, is associated with significant morbidity and mortality and is expensive, about $200,000 per patient. Genetic modification of autologous HSCT, using CD34+ cells, is emerging as a viable therapeutic alternative to allogeneic HSCT. Autologous HSCT is preferred because it is far less expensive and safer than allogeneic HSCT. One emerging approach to targeted genome modification in CD34+ cells is the use of triplex molecules, either in the form of triplex-forming oligonucleotides or peptide nucleic acids. These molecules bind to duplex DNA in a sequence-specific manner and stimulate recombination when combined with donor DNA molecules. Gene modification occurs via recruitment of the cells own DNA repair machinery, without the need for viral vectors. Viral vectors have proven problematic in such therapies. Our Specific Aim is to test the feasibility of using triplex molecules to correct one of the most prominent gene mutations in the IDUA gene, W402X. For this technology to be viable as a commercial therapy, we must demonstrate that we can achieve a success rate of at least 3% for correction of our gene target in CD34+ cells. Next, we must demonstrate that we can generate a sufficient number of monocytes from these stem cells to be utilized as part of a bridging therapy to wholesale HSCT. In Phase II, we will carry out in vivo studies to demonstrate in vivo efficacy and work on developmental and IND enabling studies to move this technology toward clinical studies. PUBLIC HEALTH RELEVANCE: Hurler Syndrome (HS), a devastating genetic disease that results in multiple organ failure with progressive deterioration in the central nervous system, is caused by mutations in a single gene that is involved in the processing of large sugar molecules within compartments in our cells. L2 Diagnostics LLC is proposing to develop a therapeutic gene targeting protocol to correct one of the most prominent mutations responsible for HS. Our protocol would repair this mutation in isolated blood stem cells from HS patients and then reintroduce the corrected cells back into the patient with the potential of having a significant impact on the symptoms of the disease.

描述（由申请人提供）：这是一份提案，旨在测试使用三分子作为治疗赫勒综合征（HS）的可行性。溶酶体贮积病（lsd），如HS，代表了具有破坏性临床特征的单基因疾病的多样化集合。lsd是由编码溶酶体酶和其他对溶酶体功能至关重要的蛋白质的基因突变引起的。HS是由- l - IDUA基因（IDUA）突变引起的。对于许多lsd，治疗包括酶替代疗法和支持性护理。然而，酶替代疗法的费用从每位患者9万美元到56.5万美元不等。另外，同种异体造血干细胞移植（HSCT）是一种非常有效的治疗方法。然而，同种异体造血干细胞移植需要hla匹配的供体，与显著的发病率和死亡率相关，而且价格昂贵，每名患者约20万美元。利用CD34+细胞对自体HSCT进行基因修饰，正成为替代同种异体HSCT的可行治疗方法。自体造血干细胞移植是首选，因为它比异体造血干细胞移植便宜得多，也安全得多。在CD34+细胞中靶向基因组修饰的一种新兴方法是使用三联体分子，以三联体形成的寡核苷酸或肽核酸的形式。这些分子以序列特异性的方式与双链DNA结合，并在与供体DNA分子结合时刺激重组。基因修饰通过细胞自身的DNA修复机制进行，而不需要病毒载体。病毒载体已被证明在这类治疗中存在问题。我们的具体目标是测试使用三重分子来纠正IDUA基因中最突出的基因突变之一W402X的可行性。为了使这项技术成为可行的商业疗法，我们必须证明我们可以在CD34+细胞中实现至少3%的成功率来校正我们的基因靶标。下一步，我们必须证明我们可以从这些干细胞中产生足够数量的单核细胞，作为大规模造血干细胞移植的桥接治疗的一部分。在第二阶段，我们将进行体内研究，以证明体内疗效，并开展发展和IND研究，使这项技术走向临床研究。