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：
7646183
负责人：
DAVID Allen WILCOX
金额：
$ 33.81万
依托单位：
MEDICAL COLLEGE OF WISCONSIN
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-04-01 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7646183
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 Cell Transplants Cells Clinical Clot retraction Coagulants Coagulation Process Codon Nucleotides 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 Outcome Patients Pharmaceutical Preparations Plasma Plasma Proteins Platelet Activation Platelet Count measurement Platelet aggregation Play Proteins Protocols documentation 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 large scale production 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）：对于血小板蛋白中遗传性分子缺陷的个体，长期出血可能会危及生命，在保持止血作用中起着复杂的作用。用正常血小板或冷冻保存血浆成分输血已用于暂时缓解间歇性出血发作，而骨髓移植已有效地用于长期纠正某些遗传性出血疾病患者。虽然，多种输血是昂贵的，并且可能导致外国血小板和凝结因子的免疫介导的破坏或失活。此外，很难找到合适的骨髓供体，并且受体也可能遭受Graft-Vs-Host疾病和移植排斥的有害影响。最近，我们通过将慢病毒载体的治疗基因转移到造血干细胞中，显示了该问题的潜在可行解决方案。该策略被证明可用于校正Glanzmann血栓症（GT）的鼠模型中的出血。具有GT的个体在整合素家族的粘附受体家族AllBBETA3中具有遗传缺陷。编码ALLB或BETA3亚基的序列中的分子异常阻止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局限于血小板谱系。当前的建议将检验以下假设：该血小板特异性病毒系统可用于对受遗传血小板缺陷影响的大动物的出血进行治疗校正。由于ALLB异常，一系列受GT影响的大比利牛斯犬将作为测试大动物血小板靶向基因治疗的可行性的模型。具体目标1将测试新型策略，以提高其将转基因表达增加到从造血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正常水平的50％。具体目标2建议在GT的犬类模型中使用临床相关的遗传转移方案来开发一种可行的人类基因治疗方法，用于遗传性血小板出血疾病。在特定的目标3中，将使用ALLB驱动的慢病毒来探索通过靶向合成和凝结因子VIII与血小板A颗粒的纠正犬类血友病A的策略。这项研究应导致为遗传性血小板出血疾病和其他止血疾病的人类基因治疗的安全有效方法的发展。