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）：涉及干细胞遗传修饰的疗法对治疗血液和免疫系统疾病的治疗具有实质性的希望。他们还面临着艰巨的障碍，限制了它们的可行性和效率，这部分与靶细胞的通道不良和培养中的干细胞存活损害有关。最近，我们进行了一系列新的观察结果，这些观察结果表明了通过造血“载体”细胞及其二次转导事件的能力延长了艾滋病毒衍生的慢病毒颗粒的细胞内保留。在鼠移植研究中，完整的颗粒的释放导致了宿主造血器官的优先颗粒传播。这些发现揭示了将颗粒捕获和活跃的载体细胞归巢组合起来，以将烯烃传递到骨髓微环境中。我们假设烟熏颗粒颗粒捕获并释放造血“载体”细胞与已建立的归巢机制相结合时，可以利用并优化，以原位骨髓干细胞的遗传修饰。我们将在三个特定目标中系统地检验这一假设。在具有干细胞缺陷的鼠移植模型中，我们将确定载体细胞通过原位粒子递送通过表型纠正长寿命干细胞的能力，评估发生这种情况的机制并确定对其他器官的传播。我们将基于载体细胞的归纳和宿主调节来测试策略，以改善载体细胞对微环境的访问和颗粒输送的效率。最后，使用生物化学测定和活细胞成像的组合，我们将研究载流子细胞内的粒子运输作为理解和操纵负责载体颗粒保留和释放的机制的手段。拟议的策略基于我们实验室的观察结果，并适应了使用细胞载体靶向肿瘤组织进行病毒颗粒递送的既定癌症基因治疗范式。通过自体细胞载体直接传递到骨髓干细胞的成功全身输送烟液颗粒将有助于克服当前对造血和免疫疾病基因治疗的障碍。没有改善组织和靶细胞的机会，基因疗法的治疗潜力在很大程度上仍未实现。因此，提出的一种新型细胞方法的遗传有效载荷的开发对于血液和免疫系统的遗传疾病的预期纠正至关重要，而且对于再生医学损伤后修复的工具至关重要。