In vivo function and tolerance to Factor VIII variants

体内功能和对因子 VIII 变体的耐受性

基本信息

批准号：
7347439
负责人：
Brendan Lee
金额：
$ 38.38万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-12-21 至 2011-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7347439
关键词：
A Mouse Acute Address Adenovirus Vector Adoptive Transfer Animal Model Antibody Formation Antigens Apolipoprotein E Autologous Autologous Dendritic Cells B-Lymphocytes Balloon Occlusion Biochemical Biomedical Engineering Blood Coagulation Disorders CCRL2 gene Calcium Canis familiaris Catheters Cell Survival Cell Therapy Cell physiology Cells Chronic Clinical Complex Complication Data Dendritic Cell Therapy Dendritic Cells Development Disease Dose Elements Exhibits Exposure to Factor VIII Gene Expression Gene Proteins Gene Transfer Generations Genes Genetic Goals Hemophilia A Hepatic Hereditary Disease Hour Human Immune response Immunity Immunologics In Vitro Inborn Genetic Diseases Infection Interleukin-10 Lead Liver Liver Circulation Maintenance Mediating Methods Modeling Modification Morbidity - disease rate Mus Patients Peripheral Personal Satisfaction Phenotype Physiologic pulse Population Heterogeneity Precipitation Process Proteins Pulse taking Rate Recombinants Replacement Therapy System T-Lymphocyte Therapeutic Index Toxic effect Transgenes Translations Variant Venous Viral Genes base calcium phosphate clinically relevant cost desire gene therapy gene transfer vector helper-dependent adenoviral vector human F8 protein improved improved functioning in vivo inhibitor/antagonist interest liver metabolism mortality mouse model novel novel strategies promoter response therapeutic protein transgene expression vector

项目摘要

DESCRIPTION (provided by applicant): Hemophilia A (HA) is a common disorder of coagulation caused by deficiency of factor VIII (FVIII). The mainstay of therapy has been replacement therapy with recombinant human FVIII. However, because of high cost, access to therapy, and inhibitory antibody formation, patients continue to suffer from significant long-term morbidity and mortality. The generation of improved versions of FVIII molecules with increased specific activity and/or improved immunological profile would improve hemophilia treatment. The expression of these new variants with liver-directed gene therapy in conjunction with strategies to tolerize patients to the therapeutic protein would constitute important clinical advances. Our development of an optimized helper-dependent adenoviral gene vector (HDV) system has enabled us to achieve long term expression of both secreted and intracellular transgenes with absent chronic toxcity and persistence of vector in small and large animal models. However, acute toxicity remains an obstacle to clinical translation. To overcome this, we have developed a novel balloon catheter based delivery system into the hepatic circulation. We have also developed a novel method for generation of tolerogenic dendritic cells based on calcium phosphate-dependent transduction of HDV expressing TGF2 and IL10. Based on these preliminary data, we will address three important questions in genetic therapy for hemophilia a) Can we increase the therapeutic index by maximizing gene expression of more bioactive FVIII variants? b) Can we increase the maximal toxic dose by overcoming the threshold/nonlinear dose response of Ad gene transfer in canine HA and achieve re-administration of the vector? c) Can we tolerize the recipient of gene transfer or h FVIII by adoptive transfer of autologous tolerogenic DCs? The overall goal of these studies is to address the major obstacle to clinical translation of adenoviral gene therapy in HA: 1) To achieve clinical correction in a large animal model of HA at a dose relevant for human translation and 2) to develop an antigen-specific tolerizing strategy for FVIII expression after gene transfer.

描述（由申请人提供）：血友病A（HA）是由于因子VIII（FVIII）缺乏引起的一种常见凝血疾病。治疗的主要是用重组人FVIII替代治疗。但是，由于成本高，接受治疗和抑制性抗体的形成，患者继续患有显着的长期发病率和死亡率。随着特定活性和/或改善免疫学特征的增加的FVIII分子的改进版本的产生将改善血友病的治疗。这些新变异与肝脏定向基因治疗的表达以及耐受患者对治疗蛋白的策略的表达将构成重要的临床进展。我们开发了优化的辅助腺病毒基因载体（HDV）系统，使我们能够实现小动物模型中慢性毒性和载体持续性的分泌和细胞内转基因的长期表达。但是，急性毒性仍然是临床翻译的障碍。为了克服这一点，我们已经开发了一种新型的基于气球导管的递送系统到肝循环中。我们还开发了一种基于表达TGF2和IL10的HDV钙依赖性转导的钙依赖性转导的耐耐产生树突状细胞的新方法。基于这些初步数据，我们将解决血友病基因治疗的三个重要问题a）我们可以通过最大化更多生物活性FVIII变体的基因表达来增加治疗指数吗？ b）我们可以通过克服犬HA中AD基因转移的阈值/非线性剂量反应并实现对载体的重新给药来增加最大毒性剂量吗？ c）我们可以通过自体耐受性DC的生育转移来耐受基因转移或h FVIII的受体吗？这些研究的总体目的是解决HA中腺病毒基因治疗临床翻译的主要障碍：1）在与人类翻译相关的大型HA动物模型中实现临床校正和2）开发抗原 - 基因转移后FVIII表达的特定耐受策略。