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.

本项目以遗传免疫缺陷疾病白细胞粘附缺陷（LAD）为模型，应用在SCOR背景下对干细胞生物学的理解进展，以增强基因向造血干细胞的转移。在LAD中，白细胞整合素CD18分子中的分子缺陷导致白细胞无法粘附血管壁并迁移到感染部位，最终导致危及生命的细菌反复发作。LAD是一个有吸引力的模型，因为：1)LAD的缺陷涉及膜受体，因此可以通过外周血白细胞的流式细胞术分析来评估基因转移的有效性；2)皮肤室法允许在体内选择性检测CD18基因校正的细胞；3) LAD重度和中度缺陷表型的存在促进了基因校正细胞输注后CD18+细胞的表型与持久性之间的相关性；4)犬类LAD （CLAD）使新的治疗方法能够在适当的大型动物模型中进行有效性和安全性测试，然后再应用于患有该疾病的人类。该项目的具体目标是：1)扩大我们目前的临床试验，利用PG13/LgCD18逆转录病毒载体将CD18体外介导的基因转移到来自LAD患者的CD34+细胞中，以包括中度缺乏和严重缺乏患者；2)利用CLAD模型和逆转录病毒介导的CD18基因转移来确定一种调节方案是否能够使植入足够的自体CD18基因校正的造血干细胞来逆转临床表型；3)基于本项目和SCOR其他项目的结果，设计未来LAD的临床基因治疗试验。在LAD和CLAD中，治疗效果将通过监测CD18+ SCOR的持续性来评估。在LAD和CLAD中，治疗效果将通过流式细胞术监测外周血中CD18+细胞的持久性、CD18+中性粒细胞向皮肤腔室的迁移以及临床表型的逆转来评估。包括SCOR中其他项目对造血干细胞理解的进展，以及该项目的结果，将加速将基础科学和临床观察转化为人类造血干细胞基因治疗的新方法。