Safe and efficient in vivo hematopoietic stem cell transduction in nonhuman primates using HDAd5/35++ vectors.
Safe and efficient in vivo hematopoietic stem cell transduction in nonhuman primates using HDAd5/35++ vectors.
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DOI：
10.1016/j.omtm.2021.12.003
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发表时间：
2022-03-10
期刊：
影响因子：
--
通讯作者：
Lieber A
中科院分区：
文献类型：
--
作者：
Li C;Wang H;Gil S;Germond A;Fountain C;Baldessari A;Kim J;Liu Z;Georgakopoulou A;Radtke S;Raskó T;Pande A;Chiang C;Chin E;Yannaki E;Izsvák Z;Papayannopoulou T;Kiem HP;Lieber A
我们在猕猴体内测试了一种新的造血干细胞转导/选择方法，使用嗜人、整合型、辅助性的腺病毒载体(HDAd5/35++)在红细胞(RBC)中表达人γ-珠蛋白，以治疗血红蛋白疾病。我们发现，HDAd5/35++载体静脉注射到粒细胞集落刺激因子(G-CSF)/AMD3100动员的动物体内后，优先转导HSCs，并将转导的细胞返回骨髓和脾。该方法耐受性良好，通过地塞米松联合白介素1和白介素6受体阻滞剂的预处理，成功地钝化了通常与静脉注射腺病毒载体有关的促炎细胞因子的激活。使用我们基于MGMTP140K的体内选择方法，所有动物的γ-珠蛋白+红细胞都增加了，水平高达90%。经过选择后，γ-珠蛋白+红细胞的百分比下降，很可能是由于对人类转基因产品的免疫反应。我们的生物分布数据表明，外周血中的γ-珠蛋白+红细胞主要来源于归巢于脾的动员的HSC。整合位点分析显示了一种多克隆模式，没有转基因整合相关的遗传毒性。这是首次在非人类灵长类动物中进行的概念验证性研究，表明在体内，HSC基因治疗在人类身上是可行的，而不需要高剂量的化疗条件和HSC移植。我们开发了一种体内造血干细胞(HSC)基因治疗方法，包括动员HSC和静脉注射亲HSC，整合HDAd5/35++载体。我们的研究表明，这种方法在恒河猴身上是安全有效的，在人类身上也是可行的。
We tested a new in vivo hematopoietic stem cell (HSC) transduction/selection approach in rhesus macaques using HSC-tropic, integrating, helper-dependent adenovirus vectors (HDAd5/35++) designed for the expression of human γ-globin in red blood cells (RBCs) to treat hemoglobinopathies. We show that HDAd5/35++ vectors preferentially transduce HSCs in vivo after intravenous injection into granulocyte colony-stimulating factor (G-CSF)/AMD3100-mobilized animals and that transduced cells return to the bone marrow and spleen. The approach was well tolerated, and the activation of proinflammatory cytokines that are usually associated with intravenous adenovirus vector injection was successfully blunted by pre-treatment with dexamethasone in combination with interleukin (IL)-1 and IL-6 receptor blockers. Using our MGMTP140K-based in vivo selection approach, γ-globin+ RBCs increased in all animals with levels up to 90%. After selection, the percentage of γ-globin+ RBCs declined, most likely due to an immune response against human transgene products. Our biodistribution data indicate that γ-globin+ RBCs in the periphery were mostly derived from mobilized HSCs that homed to the spleen. Integration site analysis revealed a polyclonal pattern and no genotoxicity related to transgene integrations. This is the first proof-of-concept study in nonhuman primates to show that in vivo HSC gene therapy could be feasible in humans without the need for high-dose chemotherapy conditioning and HSC transplantation. We have developed an in vivo hematopoietic stem cell (HSC) gene therapy approach that involves HSC mobilization and intravenous injection of HSC-tropic, integrating HDAd5/35++ vectors. Our study indicates that this approach is safe and effective in rhesus macaques and could be feasible in humans.
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影响因子：
7.5
作者：
Li, Chang;Goncalves, Kevin A.;Lieber, Andre
通讯作者：
Lieber, Andre
影响因子：
4.2
作者：
Liu, Ying;Wang, Hongjie;Lieber, Andre
通讯作者：
Lieber, Andre
影响因子：
15.9
作者：
Larochelle, Andre;Choi, Uimook;Sorrentino, Brian P.
通讯作者：
Sorrentino, Brian P.
DOI：
10.1073/pnas.2637010100
发表时间：
2004-01-06
影响因子：
11.1
作者：
Cao, YA;Wagers, AJ;Contag, CH
通讯作者：
Contag, CH
DOI：
10.3390/v13050864
发表时间：
2021-05-08
期刊：
Viruses
影响因子：
--
作者：
Davies JA;Marlow G;Uusi-Kerttula HK;Seaton G;Piggott L;Badder LM;Clarkson RWE;Chester JD;Parker AL
通讯作者：
Parker AL