Guo Qi, Parikh Keval, Zhang Jian, Brinkley Alexander, Chen Grace, Jakramonpreeya Natnicha, Zhen Anjie, An Dong Sung
UCLA AIDS Institute, UCLA, Los Angeles, CA 90024, USA.
UCLA School of Nursing, UCLA, Los Angeles, CA 90095, USA.
Mol Ther Methods Clin Dev. 2025 May 15;33(2):101486. doi: 10.1016/j.omtm.2025.101486. eCollection 2025 Jun 12.
Hematopoietic stem/progenitor cell (HSPC)-based anti-HIV-1 gene therapy holds promise to provide life-long remission following a single treatment. Here we report a multi-pronged anti-HIV-1 HSPC-based gene therapy designed to defend against and attack HIV-1 infection. We developed a lentiviral vector capable of co-expressing three anti-HIV-1 genes. Two are designed to prevent infection, including a short hairpin RNA (shRNA) (CCR5sh1005) to knock down HIV-1 co-receptor CCR5 and a membrane-anchored HIV-1 fusion inhibitor (C46). The third gene is a CD4-based chimeric antigen receptor (CAR) designed to attack HIV-1-infected cells. Our vector also includes a non-signaling truncated human epidermal growth factor receptor (huEGFRt) which acts as a negative selection-based safety kill switch against transduced cells. Anti-HIV-1 vector-transduced human CD34 HSPC efficiently reconstituted multi-lineage human hematopoietic cells in humanized bone marrow/liver/thymus (huBLT) mice. HIV-1 viral load was significantly reduced (1-log fold reduction, < 0.001) in transplanted huBLT mice. Anti-huEGFR monoclonal antibody cetuximab (CTX) administration significantly reduced huEGFRt vector-modified cells (>4-fold reduction, < 0.01) in huBLT mice. These results demonstrate that our strategy is highly effective for HIV-1 inhibition, and that CTX-mediated negative selection can deplete anti-HIV-1 vector-modified cells in the event of unwanted adverse effects in huBLT mice.
基于造血干/祖细胞（HSPC）的抗HIV-1基因疗法有望在单次治疗后实现终身缓解。在此，我们报告一种多管齐下的基于HSPC的抗HIV-1基因疗法，旨在抵御和攻击HIV-1感染。我们开发了一种能够共表达三种抗HIV-1基因的慢病毒载体。其中两种旨在预防感染，包括一种短发夹RNA（shRNA）（CCR5sh1005）以敲低HIV-1共受体CCR5和一种膜锚定的HIV-1融合抑制剂（C46）。第三种基因是一种基于CD4的嵌合抗原受体（CAR），旨在攻击HIV-1感染的细胞。我们的载体还包括一种无信号截断的人表皮生长因子受体（huEGFRt），它作为一种基于阴性选择的安全杀伤开关，用于清除转导细胞。抗HIV-1载体转导的人CD34 HSPC在人源化骨髓/肝脏/胸腺（huBLT）小鼠中有效地重建了多谱系人造血细胞。移植的huBLT小鼠中的HIV-1病毒载量显著降低（降低1个对数倍，<0.001）。在huBLT小鼠中给予抗huEGFR单克隆抗体西妥昔单抗（CTX）可显著减少huEGFRt载体修饰的细胞（降低>4倍，<0.01）。这些结果表明，我们的策略对HIV-1抑制非常有效，并且在huBLT小鼠出现不良副作用时，CTX介导的阴性选择可以清除抗HIV-1载体修饰的细胞。
