Lentiviral gene therapy for MPS type I

I 型 MPS 的慢病毒基因治疗

• 批准号: 6861180
• 负责人: Chester B. Whitley
• 金额: $ 18.22万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6861180
• 关键词: CD34 molecule; Lentivirus; NOD mouse; SCID mouse; colony stimulating factor; enzyme activity; gene delivery system; gene therapy; hematopoietic stem cells; human subject; mucopolysaccharidosis type I; recombinant proteins

Previous experience with clinical bone marrow transplantation has shown that normal marrow is capable of providing life-long systemic metabolic correction for patients with MPS diseases; however, translation of this principle to ex vivo hematopoietic stem cell gene therapy has been limited by: (a) the inability of MLV-based retroviruses to integrate into non-dividing cells; (b) high-titer vector preparations to achieve a substantial MOI; and (c) immunologic selection against transgene-expressing cells. However, recent development of third-generation lentiviral vectors suggest an approach that may obviate these problems with broad implications for many applications of ex vivo hematopoietic stem cell gene therapy. The overall objective of this project is to accomplish the requisite preclinical studies for ex vivo hematopoietic stem cell gene therapy for mucopolysaccharidosis type I (i.e., Hurler syndrome and variants). Toward this objective, Specific Aim 1 will be to construct HIV based lentiviral gene transfer systems that will be useful for treatment of human neuropathic conditions represented by the MPS diseases, and particularly targeted at transduction in human hematopoietic stem cells. Specific Aim 2 will be to evaluate the potential for intravenous administration as a means of in vivo lentiviral bone marrow stem cell transduction and metabolic correction, as well as the potential for in vivo selection of transduced cells. Follow-up evaluations will study the response to genetically-modified hematopoietic stem cells, specifically, determining the level of recombinant MPS enzyme activity achieved, the duration of expression, and clinical response. Toward an initial clinical trial, Specific Aim 3 will study vectors for expression of the normal IDUA human enzyme delivered by ex vivo lentiviral transduction of G-CSF mobilized CD34+ human peripheral blood progenitor cells after engraftment into NOD/SCID mice. In phases of this study, more complex bicistronic message vectors will be studied for the ability to enhance IDUA gene expression by means of in vivo selection of transduced cells based on antifolate drug resistance). Aliquots of G-CSF mobilized CD34+ human peripheral blood progenitor cells will be treated with IDUA vector, or IDUA-DHFR vector, and studied for (a) transduction of CFC; (b) transduction of LTCIC; and (c) transduction of marrow-repopulating cells in NOD/SCID mice. These studies are directly aimed at developing a clinical trial of ex vivo hematopoietic stem cell gene therapy for the model disease MPS I. However, the studies will have much greater significance for the broader application of lentiviral vector systems and especially for application in hematopoietic stem cell gene therapy.

以前具有临床骨髓移植的经验表明，正常骨髓能够为MPS疾病患者提供寿命的全身代谢矫正。然而，该原理向离体造血干细胞基因疗法的翻译受到限制：（a）基于MLV的逆转录病毒无法整合到非分裂细胞中； （b）高速矢量准备，以实现大量的MOI； （c）针对表达转基因细胞的免疫学选择。然而，第三代慢病毒载体的最新发展提出了一种可以消除这些问题的方法，这些方法对过体造血干细胞基因疗法的许多应用具有广泛的影响。该项目的总体目的是完成对I型粘多糖糖化病（即Hurler综合征和变体）的体内造血干细胞基因治疗的必要临床前研究。为了实现这一目标，具体目标1将是构建基于HIV的慢病毒基因转移系统，该系统将有助于治疗由MPS疾病代表的人类神经性疾病，尤其针对人类造血干细胞的转导。具体目的2将是评估静脉内给药的潜力，作为体内慢病毒骨髓干细胞转导和代谢校正的一种手段，以及在体内选择转导细胞的潜力。后续评估将研究对遗传改性的反应 造血干细胞，特别是确定重组MPS酶活性水平，表达持续时间和临床反应。在初步的临床试验中，特定的目标3将研究向载体表达通过体内慢病毒转导G-CSF传递的正常IDUA人类酶，该酶在NOD/SCID小鼠中植入了G-CSF的CD34+人类外周血祖细胞。在这项研究的阶段，将研究更复杂的双散发信息矢量，以通过基于抗叶酸耐药性在体内选择转导细胞来增强IDUA基因表达）。 G-CSF的等分试样动员CD34+人类外周血祖细胞将用IDUA载体处理，或 IDUA-DHFR载体，并研究了（a）CFC的转导； （b）LTCIC的转导； （c）点数/scID小鼠中骨髓填充细胞的转导。这些研究直接旨在开发针对模型疾病MPS的体内造血干细胞基因治疗的临床试验。但是，这些研究对于更广泛的慢病毒载体系统，尤其是在造血干细胞中的应用，将具有更大的意义。基因疗法。