HSC transduction in situ by cellular delivery of integrating viral vectors

通过整合病毒载体的细胞递送进行 HSC 原位转导

• 批准号: 8318345
• 负责人: Peter Kurre
• 金额: $ 9.28万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-05 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8318345
• 关键词: Acyclovir; Autologous; Biochemical; Biological Assay; Blood; Bone Marrow; Bone Marrow Stem Cell; CXCR; Candidate Disease Gene; Cell Survival; Cells; Defect; Development; Disease; Disorder by Site; Dose; Engraftment; Event; Face; Fanconi' s Anemia; Gene Delivery; Genetic; Goals; HIV; HIV-1; Hematological Disease; Hematopoiesis; Hematopoietic; Hematopoietic stem cells; Hereditary Disease; Homing; Immune System Diseases; Immune system; Immunofluorescence Immunologic; Improve Access; In Situ; Injection of therapeutic agent; Laboratories; Life; Marrow; Metastatic to; Mitomycins; Modality; Modeling; Modification; Mus; Mutation; Neomycin; Oncogenes; Organ; Pathway interactions; Phenotype; Process; Radiation therapy; Regenerative Medicine; Resistance; Risk; Role; Series; Site-Directed Mutagenesis; Stem cells; Testing; Therapeutic; Tissues; Transgenic Organisms; Transplantation; Tumor Tissue; Viral Vector; Virion; base; blood treatment; cell motility; cell type; cellular imaging; cellular transduction; chemokine receptor; chemotherapy; conditioning; gene therapy; genetically modified cells; hematopoietic tissue; improved; in vivo; injury and repair; insight; novel; particle; promoter; success; tool; trafficking; tumor; vector

DESCRIPTION (provided by applicant): Therapies involving the genetic modification of stem cells hold substantial promise for the treatment of blood- and immunesystem disorders. They also face daunting obstacles limiting their feasibility and efficiency, related in part to poor target cell access and compromised stem cell survival in culture. We have recently made a series of novel observations that demonstrate the prolonged intracellular retention of HIV-derived lentivector particles by hematopoietic "carrier" cells and their capacity for secondary transduction events. The release of intact particles in murine transplantation studies led to preferential particle dissemination to host hematopoietic organs. These findings reveal a unique therapeutic opportunity to combine particle capture and active carrier cell homing for the delivery of lentivector to the marrow microenvironment. We hypothesize that lentivector particle capture and release by hematopoietic "carrier" cells, when combined with established homing mechanisms, can be exploited and optimized for the genetic modification of bone marrow stem cells in situ. We will systematically test this hypothesis in three specific aims. In a murine transplantation model with a stem cell defect we will ascertain the ability of carrier cells to phenotypically correct long-lived stem cells by in situ particle delivery, evaluate the mechanism by which this occurs and determine the dissemination to other organs. We will test strategies based on carrier cell homing and host conditioning to improve carrier cell access to the microenvironment and the efficiency of particle delivery. Finally, using a combination of biochemical assays and live-cell imaging we will investigate particle trafficking within carrier cells as a means of understanding and manipulating the mechanisms responsible for vector particle retention and release. The proposed strategy is based on our laboratory's observations, and adapts the established cancer gene therapy paradigm of using cellular carriers to target tumor tissue for virus particle delivery. Successful systemic delivery of lentivector particles by autologous cellular carriers directly to bone marrow stem cells will help overcome current impediments to gene therapy for hematopoietic and immune disorders. Without improved access to tissues and target cells, the therapeutic potential of gene therapy remains largely unfulfilled. The proposed development of a novel cellular approach to the delivery of genetic payload is therefore critically important not only for the intended correction of genetic disorders of the blood and immune system, but also as a tool for post injury repair in regenerative medicine.

描述（由申请人提供）：涉及干细胞基因修饰的疗法为治疗血液和免疫系统疾病带来了巨大希望。它们还面临着限制其可行性和效率的巨大障碍，部分原因与靶细胞接触不良和培养物中干细胞存活率受损有关。我们最近进行了一系列新的观察，证明了造血“载体”细胞对 HIV 衍生的慢载体颗粒的细胞内长时间保留及其二次转导事件的能力。小鼠移植研究中完整颗粒的释放导致颗粒优先传播到宿主造血器官。这些发现揭示了将颗粒捕获和主动载体细胞归巢相结合以将慢载体递送至骨髓微环境的独特治疗机会。我们假设造血“载体”细胞捕获和释放慢载体颗粒，与已建立的归巢机制相结合，可以利用和优化骨髓干细胞的原位遗传修饰。我们将在三个具体目标上系统地检验这一假设。在具有干细胞缺陷的小鼠移植模型中，我们将确定载体细胞通过原位颗粒递送在表型上纠正长寿命干细胞的能力，评估这种情况发生的机制并确定向其他器官的传播。我们将测试基于载体细胞归巢和宿主调节的策略，以改善载体细胞进入微环境和颗粒递送的效率。最后，结合生化测定和活细胞成像，我们将研究载体细胞内的颗粒运输，作为理解和操纵载体颗粒保留和释放机制的一种手段。所提出的策略基于我们实验室的观察，并采用了已建立的癌症基因治疗范例，即使用细胞载体靶向肿瘤组织以进行病毒颗粒递送。通过自体细胞载体将慢载体颗粒成功系统地直接递送至骨髓干细胞将有助于克服目前造血和免疫疾病基因治疗的障碍。如果不改善对组织和靶细胞的接触，基因疗法的治疗潜力在很大程度上仍然无法发挥。因此，所提议的开发一种新的细胞方法来传递遗传有效负载不仅对于血液和免疫系统遗传性疾病的预期纠正至关重要，而且作为再生医学中损伤后修复的工具也至关重要。