Lentiviral Gene Therapy for Sickle Cell Disease and Immunodeficiency Disorders

描述（由申请人提供）： 该计划项目的总体目标是开发用于镰状细胞病（SCD）、Wiscott-Aldrich综合征（WAS）和X连锁严重联合免疫缺陷（SCIDXI）患者的慢病毒载体。中心统一的假设是，自失活（SIN）慢病毒载体含有绝缘子元件和适当的内部启动子将是安全和有效的治疗这些血液和免疫细胞疾病。我们的一般方法建立在上一个资助期的进展基础上，该进展导致了用于血红蛋白病和SCID-Xl的新型SEN HIV载体，其具有显著较低的细胞原癌基因意外激活倾向。我们最近还开发了一种稳定的慢病毒生产系统，这将极大地促进所有3种疾病的临床载体生产。在项目1中，我们将产生和测试用于SCD的改进载体，并开发用于扩增和转导造血干细胞的新方法。在项目2中，WAS的慢病毒载体将在临床前模型中进行有效性和安全性测试，随后将在WAS基因治疗的临床试验中进行测试。在项目3中，SCID-X1慢病毒载体的现有稳定生产细胞系将用于支持SCID-X1的两项临床试验;一项针对新诊断的患者，第二项针对先前治疗失败的年龄较大的儿童。核心提供了这3个项目所需的基础设施要素。行政核心A将为POl活动提供一般行政支持。干细胞核心B将在临床前实验和临床试验中提供处理和转导HSC的方法。载体核心C将为所有3个项目获得稳定的慢病毒生产克隆，并提供临床试验中使用的载体的生产和认证方法。免疫学核心D将提供临床研究所需的标准化免疫重建测定，并协助招募和护理方案患者。这些项目和核心具有高度的协同作用，并具有广泛的互补互动。总的来说，我们预计这项提案中的工作将产生高影响力的临床结果，这些结果将定义慢病毒载体在治疗这些疾病中的作用，并将提供有关使用慢病毒载体进行人类干细胞基因治疗的开创性信息。

DESCRIPTION (provided by applicant): The overall goal of this Program Project is to develop lentiviral vectors for use in patients with sickle cell disease (SCD), Wiscott-Aldrich Syndrome (WAS), and X-linked severe combined immunodeficiency (SCIDXI). The central unifying hypothesis is that self-inactivating (SIN) lentiviral vectors containing insulator elements and appropriate intemal promoters will be both safe and effective for the treatment of these blood and immune cell disorders. Our general approach builds upon progress during the last funding period that has led to novel SEN HIV vectors for hemoglobinopathies and SCID-Xl that have a markedly lower propensity for inadvertent activation of cellular proto-oncogenes. We have also recently developed a stable lentiviral producer system that will greatly facilitate clinical vector production for all 3 disorders. In Project 1, we will generate and test improved vectors for SCD and develop new methods for expansion and transduction of hematopoietic stem cells. In Project 2, lentiviral vectors for WAS will be tested in preclinical models for efficacy and safety and later in a clinical trial of WAS gene therapy. In Project 3, an existing stable producer cell line for a SCID-Xl lentiviral vector will be used to support two clinical trials for SCID-Xl; one for newly diagnosed patients and a second trial for older children that have failed prior therapy. The Cores provide the infrastructural elements that are necessary for these 3 projects. Administrative Core A will provide general administrative support for the POl activities. Stem Cell Core B will provide the means for processing and transducing HSCs, both in preclinical experiments and in the clinical trials. Vector Core C will derive stable lentiviral producer clones for all 3 projects and provide methods for the production and certification of vectors used in the clinical trials. Immunology Core D will provide standardized immunologic reconstitution assays required for the clinical studies and assist in the recruitment and care of protocol patients. The Projects and Cores are highly synergistic and have extensive complementary interactions. Overall, we expect that the work in this proposal will lead to high impact clinical results that will define the role of lentiviral vectors for the treatment of these diseases, and that will provide pioneering information regarding the use of lentiviral vectors for human stem cell gene therapy.