LEUKOCYTE ADHERENCE DEFICIENCY MODEL STEM CELL TRANSDUCT

白细胞粘附缺陷模型干细胞转导

• 批准号: 6652843
• 负责人: DENNIS DURAND HICKSTEIN
• 金额: $ 20.94万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6652843
• 关键词: CD34 molecule; Retroviridae; autologous transplantation; biotechnology; clinical research; clinical trials; colony stimulating factor; dogs; flow cytometry; gene expression; gene therapy; hematopoietic growth factor; hematopoietic stem cells; human subject; human therapy evaluation; inborn immunodeficiency; integrins; leukapheresis; leukocyte adhesion molecules; leukocyte disorder; nonhuman therapy evaluation; protein structure function; stem cell transplantation; tissue /cell culture; transfection /expression vector

This project uses the genetic immunodeficiency disease leukocyte adhesion deficiency or LAD as a model to which to apply advances in understanding of stem cell biology developed in the context of this SCOR to enhance gene transfer into the hematopoietic stem cell. In LAD molecular defects in the leukocyte integrin CD18 molecule result in the failure of leukocytes to adhere to the vessel wall and migrate to the site of infection culminating in recurrent episodes of life-threatening bacterial. LAD is an attractive model of the proposed studies in that: 1) the defect in LAD involves a membrane receptor, therefore efficacy of gene transfer can be assessed by flow cytometric analysis of peripheral blood leukocytes; 2) the skin chamber assay allows CD18 gene corrected cells to be selectively detected in vivo; 3) the presence of severe and moderate deficiency phenotypes of LAD facilitates correlation between the phenotype and the persistence of CD18+ cells following the infusion of gene-corrected cells; and 4) a canine form of LAD (CLAD) enables the efficacy and safety of novel therapeutic approaches to be tested in a appropriate, large-animal model prior to their application in humans with the disease. The specific aims of this project are: 1) to expand our current clinical trial of ex vivo retroviral-mediated gene transfer of CD18 into CD34+ cells from patients with LAD using the PG13/LgCD18 retroviral vector to include moderate deficiency as well as severe deficiency patients; 2) to utilize the CLAD model and retroviral-mediated gene transfer of CD18 to determine whether a conditioning regimen will enable the engraftment of sufficient autologous, CD18 gene corrected hematopoietic stem cells to reverse the clinical phenotype; and 3) to design future clinical gene therapy trials in LAD based on the results from this project and the other projects in this SCOR. In both LAD and CLAD the efficacy of therapy will be assessed by monitoring the persistence of CD18+ SCOR. In both LAD and CLAD the efficacy of therapy will be assessed by monitoring the persistence of CD18+ cells in the peripheral blood flow by flow cytometry, the migration of CD18+ neutrophils into skin chambers, and the reversal of the clinical phenotype. Including the advances in understanding of the hematopoietic stem cell from other projects in this SCOR, as well as the results from this Project, will expedite the translation of basic science and clinical observations into novel approaches to hematopoietic stem cell gene therapy in humans.

该项目使用遗传免疫缺陷疾病白细胞粘附缺乏症或LAD作为一种模型，以了解在该SCOR背景下开发的干细胞生物学方面的进步，以增强基因转移到造血干细胞中。在白细胞整联蛋白CD18分子中的LAD分子缺陷中，白细胞失去了粘附在血管壁上并迁移到感染部位，最终在危及生命的细菌的复发性发作中最终导致。 LAD是拟议研究的有吸引力的模型：1）LAD中的缺陷涉及膜受体，因此可以通过对外周血白细胞的流式细胞仪分析来评估基因转移的功效； 2）皮肤腔室测定允许在体内选择性检测到CD18基因校正细胞； 3）LAD的严重和中度缺乏表型的存在促进了在输注基因校正细胞后CD18+细胞持续性之间的相关性； 4）LAD的犬类形式（clad）使新型治疗方法的功效和安全性在适当的大动物模型中进行测试，然后才能在患有该疾病的人类中使用。该项目的具体目的是：1）扩大我们对使用PG13/LGCD18逆转录病毒载体的LAD患者的离体逆转录病毒介导的基因转移到CD34+细胞中的临床试验，以包括中度缺陷以及严重的缺乏症患者； 2）利用CD18的层层模型和逆转录病毒介导的基因转移，以确定条件方案是否能够植入足够自体的CD18基因校正的造血干细胞以逆转临床表型； 3）根据该项目的结果以及该SCOR中的其他项目，在LAD中设计未来的临床基因治疗试验。在LAD和外壳中，将通过监测CD18+ SCOR的持久性来评估治疗的功效。在LAD和外壳中，将通过流式细胞仪，CD18+中性粒细胞迁移到皮肤室中的迁移以及临床表型的逆转来评估治疗的疗效。包括了解该SCOR中其他项目的造血干细胞的进步以及该项目的结果，将加快将基础科学和临床观察结果转化为人类造血干细胞基因疗法的新方法。