Gene therapy for Fanconi anemia

范可尼贫血的基因治疗

• 批准号: 7139140
• 负责人: HANS-PETER KIEM
• 金额: $ 65.14万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-24 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7139140
• 关键词: CD34 molecule; Lentivirus; biotechnology; blood disorder chemotherapy; clinical research; clinical trial phase I; combination therapy; congenital aplastic anemia; cyclophosphamide; cytogenetics; gene delivery system; gene therapy; genetically modified animals; hematopoiesis; hematopoietic stem cells; hematopoietic tissue transplantation; human subject; human therapy evaluation; laboratory mouse; nonhuman therapy evaluation; polymerase chain reaction; stem cell transplantation; transfection /expression vector

DESCRIPTION (provided by applicant): 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 gammaretroviral 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 gammaretroviral 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) complete preclinical studies of FANCA and FANCC lentiviral vectors in human FA cells and in FANCA-/- and FABCC-/- mice and optimize procedures for clinical transduction, 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 gene- corrected FA cells, and 5) analyze lentiviral integration sites and monitor for the development of dominant clones in patients after infusion of gene-modified cells. Our ultimate goal is to improve or cure the hematopoietic symptoms in FA patients by lentivirus-mediated HSC gene transfer, and develop methodologies that would be applicable to other genetic disorders and hematological diseases.

描述（由申请人提供）：该项目的主要目的是针对Fanconi贫血患者（FA）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，对无关供体的患者的移植与显着的毒性有关，这些患者的预后不如HLA匹配的兄弟姐妹供体的患者成功。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。另外，FA细胞对低剂量的环磷酰胺高度敏感，可用于增加转基因细胞的比例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γ逆转录病毒载体的局限性，该载体需要细胞分裂和延长的细胞培养周期才能有效转导。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γ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。使用小鼠和大型动物模型，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，我们提出1）对人FA细胞和FANCA细胞中的FANCA和FANCC慢病毒载体的完整临床前研究，以及FACCA - / - 以及FABCC - / - 小鼠，并优化了临床转导的程序，2）确定被感染的基因校正的安全性和可行性细胞，3）确定基因校正细胞的持久性和体内生长优势，4）评估环磷酰胺是否可以增强基因校正的FA细胞的存活优势，5）分析慢病毒整合位点并监控为开发主要的克隆在主要的克隆中的发展。输注基因改性细胞后的患者。我们的最终目标是通过慢病毒介导的HSC基因转移改善或治愈FA患者的造血症状，并开发出适用于其他遗传疾病和血液学疾病的方法。