Therapeutic-Expression of a Platelet-Specific Integrin

血小板特异性整合素的治疗性表达

基本信息

批准号：
7318162
负责人：
DAVID Allen WILCOX
金额：
$ 35.11万
依托单位：
MEDICAL COLLEGE OF WISCONSIN
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-07-01 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7318162
关键词：
A-factor (Streptomyces)Adhesiveness Affect Allogenic Animal Model Animals Area Autologous Autologous Bone Marrow Transplantation Bernard-Soulier Syndrome Binding Bleeding time procedure Blood Cells Blood Coagulation Factor Blood Platelets Blood Vessels Bone Marrow Bone Marrow Transplantation Busulfan CD34 gene Canis familiaris Carmustine Cell Line Cells Clinical Clot retraction Coagulants Coagulation Process Codon Nucleotides Condition Contusions Cytoplasmic Granules Data Defect Development Diagnosis Disease Distal Event Factor VIII Family Fibrin Fibrinogen Foundations Future Gene Expression Gene Transfer Genes Genetic Genetic Carriers Genetic Enhancer Element Globin Goals Graft Rejection Grant Hematopoietic Hematopoietic stem cells Hemophilia A Hemorrhage Hemorrhagic Disorders Hemostatic function Histocompatibility Testing Human Immune Individual Inherited Injury Integrins Investigation Knowledge Lead Lentivirus Vector Life Marrow Mediating Megakaryocytes Methods Modeling Modification Molecular Molecular Abnormality Molecular Genetics Mus Mutagenesis Mutation Myelosuppression Other Therapy Outcome Patients Pharmaceutical Preparations Plasma Plasma Proteins Platelet Activation Platelet Count measurement Platelet aggregation Play Production Proteins Protocols documentation Purpose Recombinant Proteins Research Personnel Resistance Retroviral Vector Role Roller Bottle Site Solutions Source Subfamily lentivirinae Surface System Testing Therapeutic Thrombasthenia Transfusion Transplantation Treatment Protocols Virus Work adhesion receptor clinically relevant conditioning cost effective cytokine gene therapy gene transfer vector graft vs host disease human F8 protein in vitro Assay in vivo inhibitor/antagonist irradiation member mimetics novel strategies preclinical study preconditioning prevent programs promoter receptor recombinant antihemophilic factor VIII response therapeutic gene therapeutic target tissue culture transgene expression vector

项目摘要

DESCRIPTION (provided by applicant): Prolonged bleeding can be life-threatening for individuals with inherited molecular defects in platelet proteins that play an intricate role in maintaining hemostasis. Transfusion with normal platelets or cryopreserved plasma components has been used for temporary relief of intermittent bleeding episodes, while bone marrow transplant has been used effectively for long-term correction of some patients with inherited bleeding disorders. Although, multiple transfusions are costly and can lead to the immune mediated destruction or inactivation of foreign platelets and coagulation factors. In addition, it can be difficult to find suitable marrow donors and recipients may also suffer deleterious effects from graft-vs-host disease and graft rejection. We have recently shown a potentially feasible solution for this problem by transferring a therapeutic gene with a lentivirus vector into hematopoietic stem cells. This strategy proved useful for correction of bleeding in the murine model for Glanzmann Thrombasthenia (GT). Individuals with GT have genetic defects in a member of the integrin family of adhesion receptors, allbbeta3. Molecular abnormalities in the sequences encoding either the allb or beta3 subunit prevent GT platelets from binding fibrinogen and forming platelet aggregates. Our gene transfer vector utilizes the allb gene promoter to confine transgene expression specifically to the platelet lineage. The current proposal will test the hypothesis that this platelet-specific lentivirus system can be utilized for therapeutic correction of bleeding in large animals affected with inherited platelet defects. A line of Great Pyrenees dogs affected with GT due to an abnormality in allb will serve as a model to test the feasibility for platelet-targeted gene therapy in large animals. Specific Aim 1 will test novel strategies for their potential to increase transgene expression to 50% of normal levels on platelets derived from hematopoietic stem cells transduced with an allb gene promoter controlled lentivirus vector. Specific Aim 2 proposes to develop a feasible approach for human gene therapy for inherited platelet bleeding disorders using a clinically relevant genetic transfer protocol in a canine model for GT. In Specific Aim 3, an allb-driven lentivirus will be used to explore strategies to correct canine Hemophilia A via targeted synthesis and storage of coagulation factor VIII to platelet a-granules. This study should lead to the development of safe and effective methods for human gene therapy for inherited platelet bleeding disorders and other diseases of hemostasis.

描述（由申请人提供）：对于具有血小板蛋白遗传性分子缺陷的个体来说，长时间出血可能会危及生命，而血小板蛋白在维持止血方面发挥着复杂的作用。输入正常血小板或冷冻保存的血浆成分已被用于暂时缓解间歇性出血事件，而骨髓移植已被有效地用于长期纠正一些遗传性出血性疾病患者。然而，多次输血成本高昂，并且可能导致免疫介导的外源血小板和凝血因子的破坏或失活。此外，很难找到合适的骨髓捐献者，受者也可能遭受移植物抗宿主病和移植物排斥的有害影响。我们最近通过将治疗基因与慢病毒载体转移到造血干细胞中，展示了解决这个问题的潜在可行的解决方案。事实证明，该策略对于纠正格兰兹曼血小板无力症 (GT) 小鼠模型中的出血非常有用。 GT 患者的粘附受体整合素家族成员 allbbeta3 存在遗传缺陷。编码 allb 或 beta3 亚基的序列中的分子异常会阻止 GT 血小板结合纤维蛋白原并形成血小板聚集体。我们的基因转移载体利用 allb 基因启动子将转基因表达特异性限制在血小板谱系中。目前的提案将检验这一假设，即这种血小板特异性慢病毒系统可用于治疗性纠正患有遗传性血小板缺陷的大型动物的出血。一组因 Allb 异常而感染 GT 的大比利牛斯犬系将作为模型，测试大型动物血小板靶向基因治疗的可行性。具体目标 1 将测试新策略的潜力，以将来自用 allb 基因启动子控制的慢病毒载体转导的造血干细胞的血小板上的转基因表达增加到正常水平的 50%。具体目标 2 提议在 GT 犬模型中使用临床相关的基因转移方案，开发一种针对遗传性血小板出血性疾病的人类基因治疗的可行方法。在具体目标 3 中，将使用 allb 驱动的慢病毒探索通过将凝血因子 VIII 定向合成和储存到血小板 a 颗粒来纠正犬 A 型血友病的策略。这项研究应该会导致开发出安全有效的人类基因治疗方法，用于治疗遗传性血小板出血性疾病和其他止血疾病。