David Marion, Schiele Phillip, Monteferrario Davide, Saviane Gaëlle, Martelli Angélique E, Dupont Coralie F, Jeanneau Caroline, Marchetti Irène, Tadi Satish K, Vahldick Julia, Truong Lynn N, Zhou Yuanyue, Sauer Igor M, Schöning Wenzel, Na Il-Kang, Reik Andreas, Frentsch Marco, Rosa Maurus de la, Fenard David
Sangamo Therapeutics France, Allée de la Nertière, 06560 Valbonne, France.
BIH Center for Regenerative Therapies (BCRT), Berlin Institute of Health at Charité - Universitätsmedizin Berlin, Berlin, Germany.
Mol Ther Oncol. 2025 May 7;33(2):200989. doi: 10.1016/j.omton.2025.200989. eCollection 2025 Jun 18.
Chimeric antigen receptor T (CAR-T) therapies have shown remarkable success in treating hematological malignancies. However, effectiveness against solid tumors remains limited due to the immunosuppressive tumor microenvironment (TME), such as transforming growth factor β (TGF-β) signaling and upregulated immune checkpoints (ICs). Furthermore, identifying universal, tumor-specific targets for CAR-T cells in solid tumors is challenging, but using reinvigorated, immunosuppressive-resistant tumor-infiltrating lymphocytes (TILs) could be a promising alternative approach. Unlike nucleases, which may induce genotoxic DNA double-strand breaks, multiplexed zinc finger repressors (ZFRs) offer a safer alternative for knocking out TME-related immunosuppressive factors. We epigenetically repressed PD-1 expression both in CAR-T cells and TILs from colorectal liver metastases. PD-1 repression did not affect T cell viability, proliferation, or functionality. In a murine B cell lymphoma model, PD-1-repressed CD19-CAR-T cells exhibited enhanced anti-tumor activity and improved survival. Notably, PD-1 repression alone did not increase cytotoxicity against a PD-L1-positive colorectal cell line . To further increase anti-tumor potency in this context, ZFR-expressing lentiviral vectors (LVs) targeting PD-1 and other ICs (LAG-3, TIM-3, and TIGIT) or TGFBR2 were developed, improving significantly the cytotoxic activity in TILs. This strategy highlights the potential to enhance tumor-reactive T cells and improve anti-cancer immunotherapies by epigenetically repressing immunosuppressive factors in the TME using multiplexed ZFRs.
嵌合抗原受体T（CAR-T）疗法在治疗血液系统恶性肿瘤方面已显示出显著成效。然而，由于免疫抑制性肿瘤微环境（TME），如转化生长因子β（TGF-β）信号传导和免疫检查点（ICs）上调，其对实体瘤的疗效仍然有限。此外，在实体瘤中为CAR-T细胞识别通用的、肿瘤特异性靶点具有挑战性，但使用重新激活的、抗免疫抑制的肿瘤浸润淋巴细胞（TILs）可能是一种有前景的替代方法。与可能诱导基因毒性DNA双链断裂的核酸酶不同，多重锌指阻遏物（ZFRs）为敲除TME相关免疫抑制因子提供了一种更安全的替代方法。我们在来自结直肠癌肝转移灶的CAR-T细胞和TILs中通过表观遗传方式抑制了PD-1的表达。PD-1的抑制并未影响T细胞的活力、增殖或功能。在小鼠B细胞淋巴瘤模型中，PD-1抑制的CD19-CAR-T细胞表现出增强的抗肿瘤活性和更长的生存期。值得注意的是，单独的PD-1抑制并未增加对PD-L1阳性结直肠癌细胞系的细胞毒性。为了在这种情况下进一步提高抗肿瘤效力，开发了靶向PD-1和其他ICs（LAG-3、TIM-3和TIGIT）或TGFBR2的表达ZFR的慢病毒载体（LVs），显著提高了TILs中的细胞毒性活性。该策略突出了通过使用多重ZFRs表观遗传抑制TME中的免疫抑制因子来增强肿瘤反应性T细胞和改善抗癌免疫疗法的潜力。
