Targeted in vivo gene transfer to hematopoietic stem cells

体内靶向基因转移至造血干细胞

• 批准号: 7630452
• 负责人: Nikunj V Somia
• 金额: $ 22.65万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-01 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7630452
• 关键词: Acquired Immunodeficiency Syndrome; Applications Grants; Binding; Biological Assay; Blood; Blood Cells; Bone Marrow; CD11 Antigens; CD3 Antigens; CD34 gene; Cell Culture System; Cell Lineage; Cell Surface Receptors; Cell surface; Cells; Chimera organism; Dendritic Cells; Disease; Engineering; Evaluation; Fanconi' s Anemia; Fc Receptor; Flow Cytometry; Gene Transfer; General Population; Genes; Hematological Disease; Hematopoietic; Hematopoietic Stem Cell Mobilization; Hematopoietic System; Hematopoietic stem cells; Hemoglobinopathies; Human; Immunologic Deficiency Syndromes; In Vitro; Infection; Infusion procedures; Injection of therapeutic agent; Lentivirus Vector; Ligands; Lymphoid Cell; Marrow; Measures; Membrane Fusion; Modeling; Mus; Myelogenous; PTPRC gene; Partner in relationship; Patients; Phenotype; Phosphotransferases; Population; Protein Binding Domain; Proto-Oncogene Protein c-kit; Recombinants; Retroviral Vector; Route; Sindbis Virus; Specificity; Stem Cell Factor; Stem cells; Surface; Testing; Therapeutic; Time; Tissues; Transgenic Organisms; Transplant Recipients; Transplantation; Viral Load result; Viral Vector; base; cell type; congenic; env Gene Products; gene therapy; gene transfer vector; human stem cells; in vivo; molecular marker; mutant; peripheral blood; public health relevance; receptor; research study; targeted delivery; therapeutic gene; vector; viral vector development; virus envelope

DESCRIPTION (provided by applicant): Efficient gene transfer to human hematopoietic stem cells (HSC) has the potential to correct a number of human hematologic and non-hematologic diseases. At present, gene transfer into HSC's is typically achieved ex vivo, involving isolation of stem cells, transferring a corrective gene into the cells and re-infusion of cells into the patient. For the purpose of achieving therapeutic gene transfer, there would be great utility in a vector that could achieve targeted gene transfer in vivo to HSC. For this R21 proposal we aim to develop and test in vivo lentiviral based gene transfer vectors that will target infection through molecules that are expressed on the surface of hematopoietic stem cells. We will (i) develop lentivirus vectors for targeting infection of HSC. This will be achieved by fusing protein domains that bind cell surface receptors expressed on HCS (including CD34, and the kinase receptors c-kit and Flt3) onto a mutant Sindbis virus envelope that is severely impaired for binding its receptor but is competent for membrane fusion. These vectors will be extensively characterized using in vitro cell culture systems. We will then proceed to (ii) evaluate vectors generated for targeting infection of hematopoietic stem cells in vivo. We will investigate two routes of delivery for targeting lentiviral vectors to HSC: (a) injection into the bone marrow and (b) systemic delivery after mobilization of HSC out of the marrow into blood. These experiments will critically test the feasibility of targeting infection to HSC in vivo. Targeting vector to cells in vivo has the potential advantage of increasing gene transfer to cells while decreasing viral load. It also avoids gene transfer to cells that could impede gene therapy i.e. dendritic cells. The targeting of hematopoietic stem cells will provide proof of principle for in vivo targeting and support its use as a therapeutic approach for key hematologic disorders, such as immunodeficiencies, AIDS and Fanconi anemia. Furthermore, results from these studies will have significant implications for targeted therapeutic gene transfer using retroviral vectors in general. Public Health Relevance: This grant proposal aims to develop and test viral vectors that are engineered to infect a specific cell type in the body. Here we propose to develop these to target infection of stem cells that are responsible for making all the cells of the blood. The development of these viral vectors will enable treatment of blood disorders by direct injection of these vectors into the body. The discoveries we make can also be applied to other diseases.

描述(申请人提供)：高效的基因转移到人类造血干细胞(HSC)有可能纠正一些人类血液病和非血液病。目前，向HSC的基因转移通常是在体外实现的，包括分离干细胞，将纠正基因转移到细胞中，然后将细胞重新注入患者体内。为了达到治疗性基因转移的目的，能够在体内实现靶向基因转移的载体将具有很大的实用价值。对于这个R21方案，我们的目标是在体内开发和测试基于慢病毒的基因转移载体，该载体将通过表达在造血干细胞表面的分子来靶向感染。我们将(I)开发针对HSC感染的慢病毒载体。这将通过将结合在HCS上表达的细胞表面受体(包括CD34，以及激酶受体c-kit和Flt3)的蛋白结构域融合到突变的Sindbis病毒被膜上来实现，该病毒被膜严重受损而不能结合其受体，但具有膜融合的能力。这些载体将使用体外细胞培养系统进行广泛的表征。然后，我们将继续(Ii)评估为体内靶向感染造血干细胞而产生的载体。我们将研究靶向HSC的慢病毒载体的两种输送途径：(A)骨髓内注射和(B)动员HSC进入血液后的全身输送。这些实验将关键地测试体内靶向感染HSC的可行性。在体内靶向细胞的载体具有在减少病毒载量的同时增加向细胞的基因转移的潜在优势。它还避免将基因转移到可能阻碍基因治疗的细胞，即树突状细胞。造血干细胞的靶向将为体内靶向提供原则证据，并支持将其用作治疗免疫缺陷、艾滋病和范科尼贫血等关键血液病的方法。此外，这些研究的结果将对使用逆转录病毒载体进行靶向治疗性基因转移具有重要意义。 公共卫生相关性：这项拨款提案旨在开发和测试病毒载体，这些病毒载体经过改造后可以感染体内的特定细胞类型。在这里，我们建议开发这些细胞来针对干细胞的感染，这些干细胞负责制造血液中的所有细胞。这些病毒载体的发展将使通过将这些载体直接注射到体内来治疗血液疾病成为可能。我们的发现也可以应用于其他疾病。