Cell and Gene Therapy for Nonmalignant Blood Disorders

 描述(由申请人提供)：我们提出了一个高度协同和创新的基础和临床研究计划，旨在通过异基因造血细胞移植(HCT)和基因治疗来改善对原发免疫缺陷疾病和血红蛋白疾病患者的治疗。我们试图通过引入安全、无毒的调节疗法和避免移植物抗宿主病(GVHD)的新方法来实现这一点。该方案由四个核心支持的三个项目组成。临床项目1有三个目标，第一个目标是在接受人类白细胞抗原相合的HCT患者中完成，其余两个目标是在接受人类白细胞抗原半相合移植物的患者中完成。目的一是利用针对CD45的单抗(MAb)与放射核素α-211(211At)相偶联的靶向照射，避免当前预适应方案的毒性，特别是继发性癌症。目的二是一项多中心试验，利用自杀基因修饰的T细胞(BPX 501)和CD34+选择的人类白细胞抗原半相合干细胞进行移植，以实现低强度预适应后的植入，并避免随后的GVHD。这将确保几乎每一个患有候选疾病的患者，特别是来自少数族裔群体的患者，都可以接受异基因HCT。第三个目标是结合211At-抗CD45单抗以防止人类白细胞抗原半相合移植物的排斥反应，这一目标对镰状细胞性贫血患者尤其重要。这两个临床前项目使用的是犬的HCT模型，彼此以及与临床项目紧密结合。具体地说，项目2侧重于 四个目标。第一个是用~(211)At标记的抗CD45单抗作为丙酮酸激酶(PK)缺乏症犬的红细胞移植的条件，作为人类血红蛋白疾病的模型。第二种是用211At标记的抗CD45单抗处理X连锁严重联合免疫缺陷(SCID-X1)犬的DLA-半相合移植物，以确保持续的多系供体细胞植入。第三个目的是克服输血诱导的致敏，从而降低在DLA相合的环境中使用~(211)At标记的单抗的移植物排斥反应的风险。第四个目标是研究体外扩增的DLA相合的CD34+干细胞在没有移植物抗宿主病的情况下实现持续植入的能力。项目3将侧重于开发新的、毒性较低的调节方案，用于基因治疗、造血干细胞(HSCs)的扩增以及慢病毒载体的体内输送。我们将追求三个目标。第一项研究将评估基因修饰的造血干细胞在白花丹、硫丹或211At标记的抗CD45单抗处理后的植入情况。它还将探索自杀基因的安全开关，并进行广泛的克隆分析。目的二评价体外扩增的HSCs在非清髓性预适应后能否提高植入能力。目的评价扩增和基因修正的HSCs在犬SCID-X1和PK模型中最低限度的条件化处理后能否纠正遗传病。这项高度协同的计划提出了创新的、改变实践的基础和临床研究，旨在显著改善异基因造血细胞移植(HCT)和遗传性非恶性血液疾病(包括原发免疫缺陷和血红蛋白疾病)的基因治疗结果。开发成功的人类白细胞抗原半相合家庭成员移植方案将确保几乎每一个患有候选疾病的患者，特别是来自少数族裔群体的患者，都可以成为异基因血细胞移植的接受者。 (摘要结束) 项目1：纠正血液疾病的细胞疗法(STORB，Rainer F.)

 DESCRIPTION (provided by applicant): We propose a highly synergistic and innovative program of basic and clinical research to improve treatment of patients with primary immunodeficiency disorders and hemoglobinopathies by allogeneic hematopoietic cell transplantation (HCT) and gene therapy. We seek to accomplish this by introducing safe, non-toxic conditioning regimens and novel ways of averting graft-vs.-host disease (GVHD). The Program consists of three projects supported by four cores. The clinical Project 1 has three aims, the first to be accomplished in recipients of HLA-matched HCT and the remaining two in patients given HLA-haploidentical grafts. Aim one is to avoid toxicities of current conditioning regimens, especially secondary cancer, by using targeted irradiation with the alpha-particle emitting radionuclide astatine-211 (211At) coupled to a monoclonal antibody (MAb) directed at CD45. Aim two is a multicenter trial exploiting suicide gene modified T cells (BPX 501) given along with CD34+ selected HLA-haploidentical stem cells in order to both accomplish engraftment after reduced- intensity conditioning and avert subsequent GVHD. This would assure that virtually every patient with a candidate disease, especially patients from ethnic minority groups, can receive allogeneic HCT. The third aim is to incorporate 211At-anti-CD45 MAb to prevent rejection of HLA-haploidentical grafts, a goal particularly important for patients with sickle cell anemia. The two preclinical projects, using a canine model of HCT, are closely integrated with each other as well as with the clinical project. Specifically, Project 2 focuses on four aims. The first uses a 211At-labeled anti-CD45 radiolabeled MAb as conditioning for HCT in dogs with pyruvate kinase (PK) deficiency as model for human hemoglobinopathies. The second tackles DLA-haploidentical grafts in dogs with X-linked severe combined immunodeficiency (SCID-X1) using the 211At-labeled anti-CD45 MAb in order to assure sustained multi-lineage donor cell engraftment. The third aims to overcome transfusion- induced sensitization and consequently reduce the risk of graft rejection in the DLA-identical setting using 211At-labeled MAb. The fourth aim will study ex vivo-expanded DLA-identical CD34+ stem cells for their ability to achieve sustained engraftment without GVHD. Project 3 will focus on the development of new and less toxic conditioning regimens for gene therapy, expansion of hematopoietic stem/progenitor cells (HSCs) and on in vivo delivery of lentiviral vectors. Three aims will be pursued. The first will evaluate engraftment of gene- modified HSCs following conditioning with busulfan, treosulfan or 211At-labeled anti-CD45 MAb. It will also explore a suicide-gene safety switch and carry out extensive clonal analyses. Aim two will evaluate whether ex vivo expansion of HSCs can improve engraftment after non-myeloablative conditioning. Aim three will evaluate whether expanded and gene-corrected HSCs can correct genetic diseases in the canine SCID-X1 and PK models after minimal conditioning. The highly synergistic Program proposes innovative, practice-changing, basic and clinical research aimed at dramatically improving outcomes of allogeneic hematopoietic cell transplantation (HCT) and gene therapy for inherited nonmalignant blood diseases including primary immunodeficiencies and hemoglobinopathies. The development of successful regimens for grafts from HLA-haploidentical family members would assure that virtually every patient with a candidate disease, especially patients from ethnic minority groups, could become a recipient of allogeneic HCT. (End of Abstract) PROJECT 1: CELL THERAPY FOR CORRECTION OF BLOOD DISORDERS (STORB, RAINER F.)