LC3B upregulation by NANOG promotes immune resistance and stem-like property through hyperactivation of EGFR signaling in immune-refractory tumor cells.

Abstract

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.Abbreviations: ACTB: actin beta; ATG7: autophagy related 7; BafA1: bafilomycin A1; CASP3: caspase 3; CFSE: carboxyfluorescein succinimidyl ester; ChIP: chromatin immunoprecipitation; CI: confidence interval; CIN: cervical intraepithelial neoplasia; CSC: cancer stem cell; CTL: cytotoxic T lymphocyte; EGF: epidermal growth factor; EGFR: epidermal growth factor receptor; FIGO: International Federation of Gynecology and Obstetrics; GFP: green fluorescent protein; GZMB: granzyme B; HG-CIN: high-grade CIN; IHC: immunohistochemistry; LG-CIN: low-grade CIN; LN: lymph node; MAP1LC3B/LC3B: microtubule associated protein 1 light chain 3 beta; MCL1: myeloid cell leukemia sequence 1; MLANA/MART-1: melanoma antigen recognized by T cells 1; MUT: mutant; NANOG: Nanog homeobox; PDCD1/PD-1: programmed cell death 1; PMEL/gp100: premelanosome protein; RTK: receptor tyrosine kinase; TMA: tissue microarray; WT: wild type.

• Authors: Suyeon Kim, Hanbyoul Cho, Soon-Oh Hong, Se Jin Oh, Hyo-Jung Lee, Eunho Cho, Seon Rang Woo, Joon Seon Song, Joon-Yong Chung, Sung Wook Son, Sang Min Yoon, Yu-Min Jeon, Seunghyun Jeon, Cassian Yee, Kyung-Mi Lee, Stephen M Hewitt, Jae-Hoon Kim, Kwon-Ho Song, Tae Woo Kim
• Journal: Autophagy (Autophagy) Vol. 17 Issue 8 Pg. 1978-1997 (08 2021) ISSN: 1554-8635 [Electronic] United States
• PMID: 32762616 (Publication Type: Journal Article, Research Support, N.I.H., Intramural, Research Support, Non-U.S. Gov't)

Chemical References

• MAP1LC3B protein, human
• Map1lc3b protein, mouse
• Microtubule-Associated Proteins
• NANOG protein, human
• Nanog Homeobox Protein
• Nanog protein, mouse
• EGFR protein, human
• ErbB Receptors

Topics

• Animals
• Autophagy (genetics)
• Cell Line, Tumor
• ErbB Receptors (metabolism)
• Humans
• Mice, Inbred C57BL
• Microtubule-Associated Proteins (metabolism)
• Nanog Homeobox Protein (metabolism)
• Neoplastic Stem Cells (metabolism)
• Signal Transduction (genetics)
• Up-Regulation
• Mice