Single-dose MGTA-145/plerixafor leads to efficient mobilization and in vivo transduction of HSCs with thalassemia correction in mice
Single-dose MGTA-145/plerixafor leads to efficient mobilization and in vivo transduction of HSCs with thalassemia correction in mice
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DOI：
10.1182/bloodadvances.2020003714
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发表时间：
2021-03-02
期刊：
影响因子：
7.5
通讯作者：
Lieber, Andre
中科院分区：
文献类型：
--
作者：
Li, Chang;Goncalves, Kevin A.;Lieber, Andre
我们已经开发了一种体内造血干细胞（HSC）基因治疗方法，而不需要骨髓抑制调节和自体HSC移植。它涉及HSC动员和辅助依赖性腺病毒HDAd 5/35（++）载体系统的IV注射。目前的动员方案包括在4天内注射粒细胞集落刺激因子（G-CSF），然后给予普乐沙福/AMD 3100。我们在小鼠体内HSC转导的背景下，使用截短的GRO-β（MGTA-145;一种CXCR 2激动剂）和普乐沙福测试了一种更简单的2小时无G-CSF动员方案。MGTA-145+plerixafor组合导致HSC的稳健动员。重要的是，与G-CSF+plerixafor相比，MGTA-145+plerixafor导致白细胞增多显著减少，血清白细胞介素-6水平未升高，因此毒性可能较低。采用两种动员方案，在用O-6-苄基鸟嘌呤（O-6-BG）/BCNU进行体内选择后，在>90%的外周血单核细胞中实现了稳定的GFP标记。全基因组分析显示随机的多克隆载体整合。在地中海贫血小鼠模型中，MGTA-145+plerixafor动员后的体内HSC转导导致>95%的人γ-珠蛋白（+）红细胞，小鼠β-珠蛋白水平为36%。表型分析显示地中海贫血完全纠正。在第二次受体中保持了γ-珠蛋白标记百分比和水平，进一步证明MGTA 145 +plerixafor可动员长期再增殖的HSC。我们的研究表明，短暂暴露于MGTA-145+plerixafor作为体内HSC基因治疗应用于疾病（包括地中海贫血和镰状细胞病）的动员方案可能是有利的。
We have developed an in vivo hemopoietic stem cell (HSC) gene therapy approach without the need for myelosuppressive conditioning and autologous HSC transplantation. It involves HSC mobilization and IV injection of a helper-dependent adenovirus HDAd5/35(++) vector system. The current mobilization regimen consists of granulocyte colony-stimulating factor (G-CSF) injections over a 4-day period, followed by the administration of plerixafor/AMD3100. We tested a simpler, 2-hour, G-CSF-free mobilization regimen using truncated GRO-beta (MGTA-145; a CXCR2 agonist) and plerixafor in the context of in vivo HSC transduction in mice. The MGTA-145+plerixafor combination resulted in robust mobilization of HSCs. Importantly, compared with G-CSF+plerixafor, MGTA-145+plerixafor led to significantly less leukocytosis and no elevation of serum interleukin-6 levels and was thus likely to be less toxic. With both mobilization regimens, after in vivo selection with O-6-benzylguanine (O-6-BG)/BCNU, stable GFP marking was achieved in >90% of peripheral blood mononuclear cells. Genome-wide analysis showed random, multiclonal vector integration. In vivo HSC transduction after mobilization with MGTA-145+plerixafor in a mouse model for thalassemia resulted in >95% human gamma-globin(+) erythrocytes at a level of 36% of mouse beta-globin. Phenotypic analyses showed a complete correction of thalassemia. The gamma-globin marking percentage and level were maintained in secondary recipients, further demonstrating that MGTA145+plerixafor mobilizes long-term repopulating HSCs. Our study indicates that brief exposure to MGTA-145+plerixafor may be advantageous as a mobilization regimen for in vivo HSC gene therapy applications across diseases, including thalassemia and sickle cell disease.