Department of Biological Sciences, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Republic of Korea.
Department of Biological Sciences, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Republic of Korea; Department of Pharmaceutical Engineering, Inje University, Gimhae 50834, Republic of Korea.
J Control Release. 2022 Sep;349:367-378. doi: 10.1016/j.jconrel.2022.07.004. Epub 2022 Jul 12.
The TNF-related apoptosis-inducing ligand (TRAIL) is a promising anticancer drug candidate because it selectively binds to the proapoptotic death receptors, which are frequently overexpressed in a wide range of cancer cells, subsequently inducing strong apoptosis in these cells. However, the therapeutic benefit of TRAIL has not been clearly proven, mainly because of its poor pharmacokinetic characteristics and frequent resistance to its application caused by the activation of a survival signal via the EGF/epidermal growth factor receptor (EGFR) signaling pathway. Here, a lumazine synthase protein cage nanoparticle isolated from Aquifex aeolicus (AaLS) was used as a multiple ligand-displaying nanoplatform to display polyvalently both TRAIL and EGFR binding affibody molecules (EGFRAfb) via a SpyTag/SpyCatcher protein-ligation system, to form AaLS/TRAIL/EGFRAfb. The dual-ligand-displaying AaLS/TRAIL/EGFRAfb exhibited a dramatically enhanced cytotoxicity on TRAIL-resistant and EGFR-overexpressing A431 cancer cells in vitro, effectively disrupting the EGF-mediated EGFR survival signaling pathway by blocking EGF/EGFR binding as well as strongly activating both the extrinsic and intrinsic apoptotic pathways synergistically. The AaLS/TRAIL/EGFRAfb selectively targeted A431 cancer cells in vitro and actively reached the tumor sites in vivo. The A431 tumor-bearing mice treated with AaLS/TRAIL/EGFRAfb exhibited a significant suppression of the tumor growth without any significant side effects. Collectively, these findings showed that the AaLS/TRAIL/EGFRAfb could be used as an effective protein-based therapeutic for treating EGFR-positive cancers, which are difficult to manage using mono-therapeutic approaches.
肿瘤坏死因子相关凋亡诱导配体（TRAIL）是一种很有前途的抗癌药物候选物，因为它选择性地结合促凋亡的死亡受体，这些受体在广泛的癌细胞中经常过表达，随后在这些细胞中诱导强烈的凋亡。然而，TRAIL 的治疗益处尚未得到明确证实，主要是因为其较差的药代动力学特性，以及由于通过表皮生长因子/表皮生长因子受体（EGFR）信号通路激活生存信号，其应用经常受到抵抗。在这里，从 Aquifex aeolicus（AaLS）中分离出的乳清酸合酶蛋白笼纳米颗粒被用作多配体展示纳米平台，通过 SpyTag/SpyCatcher 蛋白连接系统多价展示 TRAIL 和 EGFR 结合适体分子（EGFRAfb），形成 AaLS/TRAIL/EGFRAfb。体外实验中，双配体展示的 AaLS/TRAIL/EGFRAfb 对 TRAIL 耐药和 EGFR 过表达的 A431 癌细胞表现出显著增强的细胞毒性，通过阻断 EGF/EGFR 结合有效破坏了 EGF 介导的 EGFR 生存信号通路，同时强烈协同激活了外在和内在凋亡途径。在体外，AaLS/TRAIL/EGFRAfb 选择性地靶向 A431 癌细胞，并在体内主动到达肿瘤部位。用 AaLS/TRAIL/EGFRAfb 治疗的 A431 荷瘤小鼠表现出肿瘤生长的显著抑制，没有任何明显的副作用。总的来说，这些发现表明 AaLS/TRAIL/EGFRAfb 可作为一种有效的基于蛋白质的治疗方法，用于治疗使用单治疗方法难以治疗的 EGFR 阳性癌症。
