Gene Therapy for Patients with Fanconi Anemia

范可尼贫血患者的基因治疗

• 批准号: 7154580
• 负责人: HANS-PETER KIEM
• 金额: $ 32.27万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-29 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7154580
• 关键词: CD34 molecule; Lentivirus; autologous transplantation; biotechnology; clinical research; congenital aplastic anemia; cooperative study; cyclophosphamide; flow cytometry; gene therapy; genetic manipulation; genetic transduction; hematopoietic growth factor; hematopoietic stem cells; hematopoietic tissue transplantation; human subject; human therapy evaluation; patient oriented research; stem cell transplantation; therapy design /development; tissue /cell culture; transfection /expression vector

The major objective of this project is to develop hematopoietic stem cell (HSC) gene therapy for patients with Fanconi anemia (FA). FA is an autosomal recessive syndrome characterized by congenital abnormalities, predisposition to malignancy, and bone marrow failure, the latter being the major cause of morbidity and mortality. Allogeneic HSC transplantation from unaffected donors is the only proven curative treatment for patients suffering from marrow failure. However, transplantation for patients with unrelated donors has been associated with significant toxicity, and outcome for these patients has been less successful than for patients with an HLA-matched sibling donor. In addition, recent data suggest that graft-versus-host disease (GVHD) increases the incidence of head and neck cancers, and leads to increased mortality in patients with FA. Gene replacement therapy using autologous hematopoietic stem cells is a potential alternative treatment modality, particularly since gene-corrected FA cells have a survival advantage. In addition, FA cells are highly sensitive to low doses of cyclophosphamide, which could be used to increase the proportion of genetically modified cells and also eliminate unmodified cells. Gene therapy for FA, however, has to date been limited by low gene transfer efficiency resulting in only transient detection of genetically modified cells and, ultimately, no clinical benefit. Part of this problem has been the limitation of oncoretroviral vectors, which require cell division and extended cell culture periods for efficient transduction. This is a particular problem for FA, since FA cells have an increased rate of apoptosis, and thus, their ability to divide and grow in culture is significantly reduced. In contrast to oncoretroviral vector, lentiviral vectors do not require cell division for transduction and can transduce stem cells even with very short transduction protocols. Using mouse and large animal models, we have recently demonstrated efficient lentiviral HSC gene transfer with an overnight transduction protocol. Thus, we propose to 1) generate lentiviral FANCA and FANCC vectors, 2) determine safety and feasibility of infusing gene-corrected cells, 3) determine persistence and in vivo growth advantage of gene-corrected cells, 4) evaluate whether cyclophosphamide can enhance the survival advantage of corrected FA cells, and 5) analyze lentiviral integration sites in patients.

该项目的主要目的是为范科尼贫血（FA）患者开发造血干细胞（HSC）基因治疗。 FA是一种常染色体隐性综合征，其特征是先天性异常，恶性肿瘤和骨髓衰竭，后者是发病率和死亡率的主要原因。未受影响的供体的同种异体HSC移植是唯一针对患有骨髓衰竭患者的治疗方法。但是，无关供体的患者的移植与明显的毒性有关，这些患者的预后不如患者成功 与HLA匹配的兄弟姐妹捐赠者。此外，最近的数据表明，移植物与宿主病（GVHD）增加了头颈癌的发生率，并导致FA患者的死亡率增加。使用自体造血干细胞替代基因替代疗法是一种潜在的替代治疗方式，尤其是因为基因校正的FA细胞具有生存优势。此外，FA细胞对低剂量的环磷酰胺高度敏感，可用于增加基因修饰的比例 细胞并消除未修饰的细胞。然而，迄今为止，FA的基因治疗受到低基因转移效率的限制，导致仅瞬时检测转基因细胞，最终没有临床益处。该问题的一部分是癌症载体的局限性，这些载体需要细胞分裂和延长的细胞培养时间才能有效转导。对于FA来说，这是一个特殊的问题，因为FA细胞的细胞凋亡率增加，因此，它们在培养中分裂和生长的能力大大降低。与癌逆转录病毒载体相反，慢病毒载体也不需要细胞分裂进行转导，即使使用非常短的转导方案也可以转导干细胞。使用鼠标和大型动物模型， 我们最近通过过夜转导方案证明了有效的慢病毒HSC基因转移。因此，我们建议1）产生慢病毒粉丝和fancc载体，2）确定注入基因校正细胞的安全性和可行性，3）确定基因校正细胞的持久性和体内生长优势，4）校正的FA细胞的生存优势，5）分析患者的慢病毒整合位点。