CORE--Canine

核心--犬类

• 批准号: 6959252
• 负责人: Timothy Charles Nichols
• 金额: $ 22.53万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6959252
• 关键词: animal colony; biomedical resource; blood chemistry; coagulation factor IX; coagulation factor VIII; disease /disorder model; dogs; gene expression; gene therapy; hemophilia As; hemophilia B; histochemistry /cytochemistry; nonhuman therapy evaluation; pathology; sex linked trait; toxicology; transfection /expression vector

The Francis Owen Blood Research Laboratory (FOBRL) will be the location of the Animal Core in this Program Project Grant (PPG) for Gene Therapy of Hemophilia. Our main objective is to use dogs with hemophilia A and hemophilia B to determine the most efficient and least toxic method of gene therapy for these inherited disorders and to monitor the expression and persistence of F.VIII and F.IX. We hypothesize that replacement of F.VIII and F.IX by gene therapy will correct the bleeder phenotype in hemophilia A and B dogs, respectively. The FOBRL, established in 1960 and dedicated to the preservation and study of severely affected animal models of bleeding disorders, has been a resource for the study of canine hemophilia A and B. Both canine hemophilia A and B in the FOBRL colony are inherited as a sex-linked recessive traits; hemophilia A is due to an intron 22 inversion and hemophilia B is due to a point mutation in the catalytic domain of F.IX. Affected dogs have no detectable antihemophilic factor functionally or antigenically. Infusion of canine or human F.VIII and F.IX concentrates corrects prolonged partial thromboplastin times and halts the severe bleeding episodes characteristic of the hemophilias. Correction of canine hemophilia A and B by liver transplantation presaged successful vector-mediated gene therapy. This strain of hemophilia B dogs is the first animal model of hemophilia to enjoy long-term benefit of gene therapy with continuous expression of canine F.IX mediated by AAV-vectors from the High laboratory for over 7 years without production of anti-canine F.IX antibodies. Some dogs have exhibited a lower bleeding rate supporting our hypothesis. Current limitations of hemophilia gene therapy include low levels of transgene expression and the relative paucity of strategies for hemophilia patients with inhibitory antibodies. The Core function, then, is to provide professional and technical personnel and laboratory facilities for the conduct of F.VIII and F.IX gene transfer and vector toxicity studies for this PPG to address these limitations through novel strategies proposed in each of the four projects. Production of the hemophilic dogs dedicated to this PPG is supported by HL63098 "Maintenance of Animal Models of Hemophilia and VWD." This core will support experimental dogs, expert personnel for liver surgery, histochemical and morphological hepatic pathology, and determination of both therapeutic levels and toxic profiles of immunosuppressive agents, transportation of dogs for experimental procedures, and storage of plasmas, gross specimens, preserved tissues, and molecular materials (DNA, RNA, constructs, cell lines, etc.) from experimental and control dogs. These hemophilic dogs with the accompanying professional and technical support are a unique resource for basic studies focused on developing methods for gene therapy of hemophilia and testing novel gene expression and transfer vectors.

弗朗西斯·欧文（Francis Owen）血液研究实验室（FOBRL）将是该计划项目赠款（PPG）的动物核心的位置，用于血友病基因治疗。我们的主要目的是使用血友病A和血友病B的狗来确定这些遗传性疾病的最有效，最不毒的基因治疗方法，并监测F.VIII和F.IX的表达和持久性。我们假设通过基因疗法替换F.VIII和F.IX将分别纠正血友病A和B犬中的流血表型。 FOBRL成立于1960年 并致力于保护和研究严重影响的出血性疾病的动物模型，一直是研究犬类血友病和B的研究资源。犬犬A和B犬A和B中的FOBRL菌落中的两种犬类血友病A和B均被遗传为是性别连接的隐性特征。血友病A是由于内含子22反转和血友病B造成的，这是由于f.ix的催化结构域中的点突变引起的。受影响的狗在功能或抗原上没有可检测到的抗生物友因因子。输注犬或人类F.VIII和F.IX浓缩物纠正了长时间的部分血小板素时间，并停止了血友病的严重出血发作。通过肝移植对犬类血友病A和B的矫正预示了成功的载体介导的基因治疗。这种血友病B狗的菌株是血友病的第一个动物模型，可享受基因治疗的长期益处，并连续表达犬F.ix 由高实验室的AAV媒介介导了7年以上，而没有产生抗官方F.IX抗体。一些狗表现出较低的出血率，支持我们的假设。血友病基因治疗的当前局限性包括低水平的转基因表达和抑制性抗体血友病患者策略的相对缺乏。因此，核心功能是为该PPG的F.VIII和F.IX基因转移和媒介毒性研究提供专业和技术人员和实验室设施，以通过四个项目中提出的新策略来解决这些局限性。 HL63098“维持血友病和VWD的动物模型”支持了致力于该PPG的血友病犬的生产。该核心将支持实验犬，肝手术专家人员，组织化学和形态学肝病理学以及 确定免疫抑制剂的治疗水平和有毒谱，狗的运输进行实验程序，以及从实验和对照狗中的血浆，总样本，保留组织和分子材料（DNA，RNA，构造，细胞系等）的存储。这些具有随附专业和技术支持的血友病犬是针对开发血友病基因疗法和测试新基因表达和转移载体的基础研究的独特资源。