Cell selection strategies for the gene therapy of the beta-hemoglobinopathies

β-血红蛋白病基因治疗的细胞选择策略

基本信息

批准号：
7810543
负责人：
Philippe Leboulch
金额：
$ 41.57万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-04-01 至 2012-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7810543
关键词：
Animal Experimentation Animal Model Autologous Benign Biological Bone Marrow Bone Marrow Purging CD34 gene Cell Maintenance Cell membrane Cells Chimeric Proteins Chimerism Clinical Trials Clinical trial protocol document Data Development Disease Erythrocytes Erythrocytoses Erythroid Erythropoietin Erythropoietin Receptor Evaluation France Genes Genetic Materials Globin Goals HIV Hematological Disease Hematopoietic Hematopoietic stem cells Hemoglobinopathies Hereditary Disease High Pressure Liquid Chromatography Human In Vitro Inherited Lentivirus Vector Longevity Macaca fascicularis Macaca mulatta Measures Membrane Modeling Monkeys Mus NGFR Protein Natural History Nerve Growth Factor Receptors Oncogenic Papio Patients Phase Population Procedures Property Regimen Residual state Risk Self-control as a personality trait Sickle Cell Anemia System Testing Thalassemia Therapeutic Time Transplantation Treatment Efficacy Variant base cellular transduction clinical application conditioning design expression vector gene therapy genetic variant in vivo lentivirally transduced magnetic field mouse model nonhuman primate novel novel strategies receptor research study vector

项目摘要

DESCRIPTION (provided by applicant): The beta-hemoglobinopathies are the most prevalent genetic disorders worldwide and serve as an important paradigm for the development of safe and effective approaches to hematopoietic gene therapy. In the past several years, substantial advances have been made towards this goal, from original vector design to the sustained correction of relevant mouse models, culminating with the first Phase I/II clinical trial of a lentiviral vector aimed at the gene therapy of a genetic disease. While the first patient treated show sustained expression of the transferred globin gene 4 months post-transplantation, the latest time assessed, we are now asking whether therapeutic globin expression within virtually all red blood cells can be safely achieved in sub-myeloablated recipients even when only partial chimerism with transduced hematopoietic stem cells (HSCs) would be obtained by current measures. In Specific Aim 1, we will devise an ex-vivo selection procedure for genetically corrected HSCs by means of a novel nerve growth factor receptor (NGF-R) variant devoid of residual activity and compatible with clinically-applicable bulk cell purification in a magnetic field. Importantly, we will assess whether the previously unavoidable loss of HSC content that occurs in vitro during such a procedure can be alleviated by means of novel HOX fusion proteins that penetrate cell membranes directly to induce HSC maintenance and expansion without the potentially oncogenic risk posed by the transfer of genetic material. In Specific Aim 2, we will investigate whether self-controlled in vivo amplification of genetically corrected red blood cells can be provided by co-expression of a natural erythropoietin (Epo) receptor variant with enhanced sensitivity to endogenous Epo. The rationale is based on the benign natural history of the familial erythrocytosis caused by this genetic variant and extensive preliminary data. In Specific Aim 3, we will turn to a non-human primate, Macaca fascicularis, whose cell transducibility by human lentiviral vectors is similar to that of human cells. This model will circumvent the low permissivity of most non-human primates for HIV vectors and make thus possible the critical evaluation of novel globin lentiviral vectors and aforementioned transplantation strategies in a large animal model closely related to humans.

描述（由申请人提供）：β-血红蛋白病是全世界最普遍的遗传性疾病，是开发安全有效的造血基因治疗方法的重要范例。在过去的几年里，从最初的载体设计到相关小鼠模型的持续修正，朝着这一目标取得了实质性进展，最终以针对遗传性疾病基因治疗的慢病毒载体进行了第一个I / II期临床试验。疾病。虽然第一位接受治疗的患者在移植后 4 个月（评估的最新时间）显示出转移的珠蛋白基因的持续表达，但我们现在想知道是否可以在亚清髓性受者中安全地实现几乎所有红细胞内的治疗性珠蛋白表达，即使仅当通过目前的措施将获得与转导的造血干细胞（HSC）的部分嵌合。在具体目标 1 中，我们将通过一种新型神经生长因子受体 (NGF-R) 变体设计一种基因校正 HSC 的离体选择程序，该变体没有残留活性，并且与临床适用的磁场中批量细胞纯化兼容。重要的是，我们将评估是否可以通过直接穿透细胞膜诱导 HSC 维持和扩增的新型 HOX 融合蛋白来减轻在此类手术过程中在体外发生的先前不可避免的 HSC 含量损失，而不会产生潜在的致癌风险。遗传物质的转移。在具体目标 2 中，我们将研究是否可以通过对内源性 Epo 敏感性增强的天然促红细胞生成素 (Epo) 受体变体的共表达来提供基因校正红细胞的自控体内扩增。其基本原理是基于由这种遗传变异引起的家族性红细胞增多症的良性自然史和大量的初步数据。在具体目标 3 中，我们将转向非人类灵长类动物食蟹猴，人类慢病毒载体对其细胞的转导能力与人类细胞相似。该模型将规避大多数非人类灵长类动物对 HIV 载体的低容许度，从而使得在与人类密切相关的大型动物模型中对新型珠蛋白慢病毒载体和上述移植策略进行严格评估成为可能。