Treatment Leukocyte Adhesion Deficiency by Foamy Virus

泡沫病毒治疗白细胞粘附缺陷

• 批准号: 7128279
• 负责人: David W Russell
• 金额: $ 37.23万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-01 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7128279
• 关键词: CD antigens; CD34 molecule; DNA damage; Retroviridae; T lymphocyte; autosomal recessive trait; biotechnology; cell population study; disease /disorder model; dogs; gene therapy; genetic promoter element; hematopoietic tissue transplantation; immunodeficiency; leukemia; leukocyte adhesion molecules; longitudinal animal study; neutrophil; nonhuman therapy evaluation; polymerase chain reaction; provirus; stem cell transplantation; therapy adverse effect; therapy design /development; transfection /expression vector; virus integration

DESCRIPTION (provided by applicant): This proposal will develop foamy virus (FV) vectors for the treatment of leukocyte adhesion deficiency (LAD). In LAD, mutations in the CD18 gene prevent the migration of white blood cells (leukocytes) into tissues, resulting in life-threatening bacterial infections. Transplantation of autologous hematopoietic stem cells (HSCs) transduced by CD18-expressing vectors could cure this disease. However, human and large animal HSCs are difficult to transduce efficiently, and integrating vectors based on murine leukemia virus (MLV) have been shown to cause leukemia by activating nearby cellular proto-oncogenes. FVs are retroviruses and vectors based on them are able to efficiently deliver genes to HSCs. They are not pathogenic so FV vectors may be safer than other types of retroviral vectors. Here FV vectors will be used to deliver the canine CD18 gene to the HSCs of dogs with canine LAD (CLAD). These HSCs will then be transplanted into CLAD dogs, where they will differentiate into mature blood cells that express CD18. If adequate CD18 levels are obtained, the transplanted dogs should improve clinically and avoid an early death due to infections. The data will be used to determine the efficiency of transferring genes into canine HSCs with FV vectors. Integration site analysis will establish how many different transduced cell clones contributed to blood formation, and whether there was selective expansion of particular clones. The transplanted animals will be followed long-term to look for potential side effects, including the development of leukemia. Different conditioning regimens used to promote engraftment of transplanted cells will be tested. FV vectors that use different promoters will be evaluated. FV vectors will be compared to similar lentiviral vectors that express CD18 to determine the best type of vector for HSC gene therapy of LAD. These experiments will determine if FV vectors can efficiently transfer genes to canine bone marrow stem cells, and if FV vectors expressing CD 18 can cure CLAD. This could lead to the development of new treatments for LAD, as well as other genetic or acquired blood diseases, which could improve the health of many Americans. If shown to be safe and effective, the FV vectors tested in the proposed experiments could then be used in human clinical trials to treat LAD.

描述(由申请人提供)： 这项提议将开发泡沫病毒(FV)载体用于治疗白细胞黏附缺陷(LAD)。在LAD中，CD18基因的突变阻止了白细胞(白细胞)迁移到组织中，导致威胁生命的细菌感染。CD18表达载体转导的自体造血干细胞(HSCs)移植可治愈该病。然而，人和大动物的HSCs很难有效地转导，基于小鼠白血病病毒(MLV)的整合载体被证明通过激活附近的细胞原癌基因而导致白血病。FV是一种逆转录病毒，基于它们的载体能够有效地将基因输送到HSCs。它们不是致病性的，因此FV载体可能比其他类型的逆转录病毒载体更安全。在此，我们将利用FV载体将犬CD18基因导入患有犬LAD(CLAD)的犬的HSCs。然后，这些造血干细胞将被移植到穿着衣服的狗身上，在那里它们将分化为表达CD18的成熟血细胞。如果获得足够的CD18水平，移植犬的临床情况应该会有所改善，避免因感染而过早死亡。这些数据将被用来确定用FV载体将基因转移到犬HSCs的效率。整合位点分析将确定有多少不同的转导细胞克隆有助于造血，以及是否存在特定克隆的选择性扩增。将对移植的动物进行长期跟踪，以寻找潜在的副作用，包括发展成白血病。用于促进移植细胞植入的不同调节方案将进行测试。使用不同启动子的FV载体将被评估。将FV载体与表达CD18的类似慢病毒载体进行比较，以确定用于LAD HSC基因治疗的最佳载体类型。这些实验将确定Fv载体是否能有效地将基因转移到犬骨髓干细胞，以及表达CD18的Fv载体是否能治愈包膜。这可能会导致LAD以及其他遗传性或获得性血液疾病的新疗法的开发，这可能会改善许多美国人的健康。如果被证明是安全和有效的，在拟议的实验中测试的FV载体随后可以用于治疗LAD的人体临床试验。