Stem Cell Gene Therapy for Hemaglobinopatathies

为了使血红蛋白疾病的基因治疗成为现实，必须开发能够高水平表达、能有效转导造血干细胞且不会导致疾病的基因转移。该项目的目标是进行与这三个挑战相关的研究。具体目标1是针对我们对染色质绝缘体的持续研究，染色质绝缘体是可以增加基因转移载体的表达和可能的安全性的元件。这包括在灵长类动物移植模型中测试原型绝缘体的活性，以及克隆和鉴定新的染色质绝缘体。具体目标2针对的是表征新的染色质绝缘体。具体目标2旨在确定临床相关造血细胞中载体整合的位置，特别强调确定对开放染色质的偏好，开放染色质是一种与活性基因相关的独特染色体结构。具体目标3针对的是在临床相关的灵长类移植模型中，目前可用的和由计划项目参与者开发的珠蛋白基因病毒载体的特性。这包括基于各种重组病毒的载体，如肿瘤逆转录病毒、慢病毒、泡沫病毒和混合腺病毒/腺相关病毒。与该计划项目的其他项目相结合，这些具体目标将提高珠蛋白基因重组病毒载体的安全性和有效性，并在将其用于临床试验之前，为这些载体的潜在风险和好处提供关键的见解。

In order for gene therapy of the hemoglobinopathies to become a reality, gene transfer must be developed that are capable of high-level expression, that can efficiently transduce hematopoietic stem cells, and do not cause disease. The goal of this project is to perform studies relevant to these three challenges. Specific Aim 1 is directed toward our continued investigations of chromatin insulators, elements that can increase both the expression and possibly the safety of gene transfer vectors. This includes testing the activity of a prototypic insulator in a primate transplantation model, and cloning and characterizing new chromatin insulators. Specific Aim 2 is directed toward characterizing new chromatin insulators. Specific Aim 2 is directed toward characterizing the sites of vector integration in clinically, relevant hematopoietic cells, with a special emphasis on identifying any preference for open chromatin, a unique chromosomal structure associated with active genes. Specific Aim 3 is directed toward characterizing the properties of virus vectors for globin gene available now and to be developed by the Program Project participants, in a clinically relevant primate transplantation model. This includes vectors base on a variety of recombinant viruses such as oncoretroviruses, lentiviruses, foamy viruses, and hybrid adeno / adeno-associated viruses. Combined with the other Projects of this Program Project, these specific aims will improve both the safety and efficacy of recombinant virus vectors for globin genes, and provide critical insights into the potential risks and benefits of these vectors prior to their use in clinical trials.