Immune selection drives tumor cells to acquire refractory phenotypes. We previously demonstrated that cytotoxic T lymphocyte (CTL)-mediated immune pressure enriches NANOG⁺ tumor cells with stem-like and immune-refractory properties that make them resistant to CTLs. Here, we report that the emergence of refractory phenotypes is highly associated with an aberrant macroautophagic/autophagic state of the NANOG⁺ tumor cells and that the autophagic phenotype arises through transcriptional induction of MAP1LC3B/LC3B by NANOG. Furthermore, we found that upregulation of LC3B expression contributes to an increase in EGF secretion. The subsequent hyperactivation of EGFR-AKT signaling rendered NANOG⁺ tumor cells resistant to CTL killing. The NANOG-LC3B-p-EGFR axis was preserved across various types of human cancer and correlated negatively with the overall survival of cervical cancer patients. Inhibition of LC3B in immune-refractory tumor models rendered tumors susceptible to adoptive T-cell transfer, as well as PDCD1/PD-1 blockade, and led to successful, long-term control of the disease. Thus, our findings demonstrate a novel link among immune-resistance, stem-like phenotypes, and LC3B-mediated autophagic secretion in immune-refractory tumor cells, and implicate the LC3B-p-EGFR axis as a central molecular target for controlling NANOG⁺ immune-refractory cancer.

免疫选择驱使肿瘤细胞获得难治性表型。我们以前证明了细胞毒性T淋巴细胞（CTL）介导的免疫压力使NANOG ⁺肿瘤细胞具有干样和免疫不应性特性，从而使其对CTL具有抗性。在这里，我们报告难治性表型的出现与NANOG ⁺肿瘤细胞的异常自噬/自噬状态高度相关，并且自噬表型是由NANOG通过转录诱导MAP1LC3B / LC3B产生的。此外，我们发现LC3B表达的上调有助于EGF分泌的增加。EGFR-AKT信号转导的随后过度激活导致NANOG ⁺对CTL杀伤具有抗性的肿瘤细胞。NANOG-LC3B-p-EGFR轴在各种类型的人类癌症中均得到保留，并且与宫颈癌患者的整体生存呈负相关。在免疫难治性肿瘤模型中抑制LC3B使得肿瘤易于过继性T细胞转移以及PDCD1 / PD-1阻断，从而成功，长期地控制了该疾病。因此，我们的发现证明了免疫抵抗性肿瘤细胞中免疫抵抗，茎样表型和LC3B介导的自噬分泌之间存在新的联系，并暗示LC3B-p-EGFR轴作为控制NANOG ⁺免疫的中心分子靶标-难治性癌症。