CORRECTION OF CONGENITAL DISEASES BY STEM CELL THERAPY

(改编自申请人的摘要)不包括异基因骨髓 同种异体骨髓移植治疗儿童先天性疾病 根据定义，干细胞是治标不治本的。患者的临床病程 (PTS)大多数此类疾病的典型特征是慢性发病率 继发于疾病和后遗症的病理生理学 可供选择的治疗方法。这个项目的目标是翻译 在以前的所有章节中进行的基础和临床前研究 当他们在临床实验中达到高潮时，设计的目的是纠正先天性 利用干细胞疗法治疗疾病。两种方法将是 具有利用全能的共同特征的使用 干细胞作为一种载体在足够的时间内产生缺失的蛋白质 逆转临床表型的数量。第一种方法依赖于 关于包括干细胞在内的整个造血系统的替换 手机。具体地说，我们建议将allBMT的可用性扩展为 促进使用单倍体相合家系的治疗选择 作为捐赠者的成员。第二种方法基于这样的原则 有可能稳定地引入功能版本的 将有缺陷的基因植入全能干细胞。具体来说，我们建议 PTS体外首次移植修饰原始造血细胞的研究 利用病毒载体对功能基因进行编码，然后尝试重组 造血和正常的基因表达和功能。要实现这些目标 目标方面，我们提出了两个具体目标。首先，我们计划进行 临床试验，以证明其可行性、安全性和有效性 设计用来麻醉或删除供体BM T细胞的操作 特异性宿主同种异体抗原改善创伤后移植物抗宿主病 单倍体相合的骨髓移植。在这些研究中，我们计划立即进入 临床尝试诱导供者骨髓移植对宿主同种异体抗原的无反应 符合条件的先天疾病患者中的细胞 单倍体匹配的骨髓移植。正如临床前和临床研究所表明的那样，我们 是否会尝试克隆删除宿主特异的供体T细胞 符合条件的恶性血液病患者中的同种异体抗原 单倍体匹配的骨髓移植，然后将这种方法扩展到选定的PT 先天疾病。第二，我们计划进行一系列试点 临床研究以确定该疗法的可行性和安全性 将导致转让的长期表达的方法 人类造血干细胞中的基因。为了确定是否 这种操作使干细胞对高剂量化疗产生抵抗力 在复发时，我们计划将MDR-1基因引入到 接受1)allBMT后复发风险较高的PTS的供者骨髓 恶性血液病；2)卵巢癌自体骨髓移植。 最终，我们将尝试治愈造血干细胞疾病 通过基因疗法。这两种并行方法的发展 在最终的分诊治疗中应该产生最大的灵活性 具体的病症和精神障碍。这些研究的进展是高度的 依赖于之前所有项目的成功。

(Adapted from the applicant's abstract) Excluding allogeneic bone marrow transplantation (alloBMT), the treatment of congenital diseases of the stem cell is by definition palliative. The clinical course of patients (pts) with most of these disorders is typified by chronic morbidity secondary to both the pathophysiology of the disorder and the sequelae of available therapy. The goal of this project is the translation of the basic and preclinical studies undertaken in all previous sections as they culminate in clinical experiments designed to correct congenital diseases by utilizing stem cell therapy. Two approaches will be employed which have the common feature of utilization of the totipotent stem cell as a vehicle to produce a missing protein in sufficient quantities to reverse a clinical phenotype. The first approach relies on replacement of the entire hematopoietic system, including the stem cell. Specifically, we propose to extend the availability of alloBMT as a therapeutic option by facilitating the use of haploidentical family members as donors. The second approach is based upon the principle that it may be possible to stably introduce a functional version of a defective gene into the totipotent stem cell. Specifically, we propose to first modify primitive hematopoietic cells of pts ex vivo by transfer of functional genes using viral vectors and then attempt to reconstitute hematopoiesis and normal gene expression and function. To achieve these objectives, we propose two specific aims. First, we plan to undertake clinical trials to demonstrate the feasibility, safety and efficacy of manipulations designed to either anergize or delete donor BM T cells specific for host alloantigen to ameliorate GVHD in pts undergoing haploidentical BMT. In these studies, we plan to immediately enter the clinic attempting to induce anergy to host alloantigen in donor BMT cells in selected pts with congenital diseases eligible for haplomismatched BMT. As preclinical and clinical studies dictate, we will pilot attempts to clonally delete donor T cells specific for host alloantigens in selected pts with hematologic malignancies eligible for haplomismatched BMT and then extend this approach to selected pts with congenital diseases. Second, we plan to undertake a series of pilot clinical studies to establish the feasibility and safety of methodologies which will lead to long term expression of transferred genes in human hematopoietic stem cells. In order to determine whether this manipulation confers stem cell resistance to high dose chemotherapy at the time of relapse, we plan to introduce the MDR-1 gene into the donor BM of pts at high risk for relapse after undergoing 1) alloBMT for hematologic malignancies and 2) autologous BMT for ovarian carcinoma. Ultimately, we will attempt to cure hematopoietic stem cells diseases by gene therapy. The development of these two parallel approaches should yield the most flexibility in eventual triage of therapy for specific pts and disorders. The evolution of these studies is highly dependent on the success of all previous projects.