此前，我们报道了人类Aγ珠蛋白基因慢病毒（LV）GbG的开发，该慢病毒表达高水平的HbF以纠正伯克利SCA小鼠模型中的镰状细胞贫血（SCA）表型，然后修饰了γ珠蛋白通过用 Aγ-珠蛋白基因中的天冬氨酸取代密码子 16 处的甘氨酸来生成 GbGM LV。在本研究中，我们评估了携带 GbGM LV 的人 Aγ 球蛋白基因在野生型小鼠中进行 GbGM 修饰的造血干细胞 (HSC) 初次和二次移植超过 18 个月后的长期安全性。通过监测对体重、血液学、组织病理学、恶性肿瘤形成和存活的影响来评估 GbGM 骨髓移植的安全性。移植模拟转导和脾病灶形成病毒（SFFV）γ-逆转录病毒载体（RV）转导的HSC的小鼠分别作为阴性和阳性对照。模拟组、GbGM LV 组和 SFFV RV 组的平均供体细胞植入情况相当。与对照组相比，接受 GbGM 治疗的小鼠的体重、临床体征、免疫表型或组织病理学没有显着差异。流式细胞术分析和原位杂交证明，四只 SFFV RV 治疗的小鼠（但没有 GbGM 治疗的小鼠）出现了供体来源的载体阳性淋巴瘤。这些结果强调了在小鼠初次和二次移植后长期随访中施用 GbGM LV 修饰的 HSC 的安全性。该数据支持在美国启动 1/2 期首次人体 SCA 临床试验。版权所有：© 2024 Shadid 等人。这是一篇根据知识共享署名许可条款分发的开放获取文章，允许在任何媒体上不受限制地使用、分发和复制，前提是注明原始作者和来源。

Previously, we reported the development of a human Aγ-globin gene lentivirus (LV), GbG, which expresses high levels of HbF to correct the sickle cell anemia (SCA) phenotype in the Berkeley SCA mouse model, and then modified the γ-globin gene by substituting glycine at codon 16 with aspartic acid in the Aγ-globin gene to generate GbGM LV. In the present study, we evaluated the long-term safety of human Aγ-globin gene carrying GbGM LV in wild-type mice after primary and secondary transplants of GbGM-modified hematopoietic stem cells (HSC) over 18 months. The safety of the GbGM bone marrow transplant was assessed by monitoring the effects on body weight, hematology, histopathology, malignancy formation, and survival. Mice transplanted with Mock-transduced and spleen focus forming virus (SFFV) γ-retroviral vector (RV)-transduced HSC served as negative and positive controls, respectively. The mean donor-cell engraftment was comparable across Mock, GbGM LV, and SFFV RV groups. There were no significant differences in body weight, clinical signs, immunophenotype, or histopathology in the GbGM-treated mice compared to controls. Four SFFV RV-treated mice, but none of the GbGM-treated mice, developed donor-derived, vector-positive lymphomas as demonstrated by flow cytometry analysis and in situ hybridization. These results highlight the safety of the administration of GbGM LV-modified HSC with long-term follow-up after primary and secondary transplants in mice. This data supported the initiation of phase 1/2 first-in-human SCA clinical trial in the United States.Copyright: © 2024 Shadid et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.