Han Jing-Fen, El-Amouri Salim S, Dai Mei, Cao Phuong, Pan Dao
Molecular and Cell Therapy Program, Division of Experimental Hematology and Cancer Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
Department of Pediatrics, University of Cincinnati School of Medicine, Cincinnati, OH 45219, USA.
Mol Ther Methods Clin Dev. 2018 Oct 10;11:52-64. doi: 10.1016/j.omtm.2018.10.001. eCollection 2018 Dec 14.
Novel strategies are needed to solve the conundrum of achieving clinical efficacy with high vector copy numbers (VCNs) in hematopoietic stem cells (HSCs) while attempting to minimize the potential risk of oncogenesis in lentiviral vector (LV)-mediated gene therapy clinical trials. We previously reported the benefits of reprogramming erythroid-megakaryocytic (EMK) cells for high-level lysosomal enzyme production with less risk of activating oncogenes in HSCs. Herein, using a murine model of mucopolysaccharidosis type I (MPS I) with a deficiency of α-L-iduronidase (IDUA), we sought to determine the transgene minimum effective doses (MEDs) in major organs, and if a transient increase of IDUA-containing red blood cells and platelets by repeated phlebotomy would provide further therapeutic benefits in diseased mice after EMK-restricted LV-mediated gene therapy. The MEDs for complete metabolic correction ranged from 0.1 to 2 VCNs in major visceral organs, which were dramatically reduced to 0.005-0.1 VCN by one cycle of stress induction and were associated with a further reduction of pathological deficits in mice with 0.005 VCN. This work provides a proof of concept that transiently stimulating erythropoiesis and thrombopoiesis can further improve therapeutic benefits in HSC-mediated gene therapy for MPS I, a repeatable and reversible approach to enhance clinical efficacy in the treatment of lysosomal storage diseases.
在慢病毒载体(LV)介导的基因治疗临床试验中,需要新的策略来解决造血干细胞(HSC)中高载体拷贝数(VCN)实现临床疗效,同时尽量降低致癌风险这一难题。我们之前报道了重编程红系-巨核系(EMK)细胞以实现高水平溶酶体酶产生且激活HSC中致癌基因风险较低的益处。在此,我们使用I型黏多糖贮积症(MPS I)的小鼠模型,该模型缺乏α-L-艾杜糖醛酸酶(IDUA),旨在确定主要器官中的转基因最低有效剂量(MED),以及通过重复放血使含IDUA的红细胞和血小板短暂增加是否会在EMK限制的LV介导的基因治疗后为患病小鼠带来进一步的治疗益处。主要内脏器官中实现完全代谢校正的MED为0.1至2个VCN,通过一个应激诱导周期可显著降至0.005 - 0.1个VCN,且与0.005个VCN的小鼠病理缺陷进一步减少相关。这项工作提供了一个概念验证,即短暂刺激红细胞生成和血小板生成可进一步改善HSC介导的MPS I基因治疗的治疗益处,这是一种在溶酶体贮积病治疗中增强临床疗效的可重复且可逆的方法。