Hypoxia‐induced miR‐210‐3p expression in lung adenocarcinoma potentiates tumor development by regulating CCL2 mediated monocyte infiltration

• Published in: Molecular oncology 2024-05, Vol.18 (5), p.1278-1300
• Main Authors: Arora, Leena, Patra, Debarun, Roy, Soumyajit, Nanda, Sidhanta, Singh, Navneet, Verma, Anita K., Chakraborti, Anuradha, Dasgupta, Suman, Pal, Durba
• Format: Article
• Language: English
• Subjects: 3' Untranslated regions; Adenocarcinoma; Adenocarcinoma of Lung - genetics; Adenocarcinoma of Lung - metabolism; Adenocarcinoma of Lung - pathology; Animals; Antibodies; CCL2; Cell culture; Cell Hypoxia - genetics; Cell Line, Tumor; Chemokine CCL2 - genetics; Chemokine CCL2 - metabolism; Chemokines; Cobalt; Deoxyribonucleic acid; Development and progression; DNA; Down-regulation; Gene expression; Gene Expression Regulation, Neoplastic; Health aspects; HIF‐1Α; Humans; Hypoxia; Hypoxia-inducible factor 1; Hypoxia-Inducible Factor 1, alpha Subunit - genetics; Hypoxia-Inducible Factor 1, alpha Subunit - metabolism; Hypoxia-inducible factor 1a; Immunomodulation; Infiltration; Leukocyte migration; Ligands; LUAD; Lung cancer; Lung Neoplasms - genetics; Lung Neoplasms - metabolism; Lung Neoplasms - pathology; Lungs; Macrophages; Metastases; MicroRNA; MicroRNAs; MicroRNAs - genetics; MicroRNAs - metabolism; miRNA; miR‐210‐3p; Molecular modelling; Monocyte chemoattractant protein 1; monocyte infiltration; Monocytes; Monocytes - metabolism; Monocytes - pathology; mRNA; Olefins; Phenotypes; Proteins; Scientific equipment and supplies industry; Spheroids; Transcription factors; Tumor cell lines; Tumors; Zebrafish
• ISSN: 1574-7891; 1878-0261
• DOI: 10.1002/1878-0261.13260

In most cancers, tumor hypoxia downregulates the expression of C‐C motif chemokine 2 (CCL2), and this downregulation has been implicated in monocyte infiltration and tumor progression; however, the molecular mechanism is not yet clear. We compared noncancerous and lung‐adenocarcinoma human samples for hypoxia‐inducible factor 1‐alpha (HIF‐1A), microRNA‐210‐3p (mir‐210‐3p), and CCL2 levels. Mechanistic studies were performed on lung adenocarcinoma cell lines and 3D tumor spheroids to understand the role of hypoxia‐induced miR‐210‐3p in the regulation of CCL2 expression and macrophage polarization. HIF‐1Α stabilization increases miR‐210‐3p levels in lung adenocarcinoma and impairs monocyte infiltration by inhibiting CCL2 expression. Mechanistically, miR‐210‐3p directly binds to the 3′untranslated region (UTR) of CCL2 mRNA and silences it. Suppressing miR‐210‐3p substantially downregulates the effect of hypoxia on CCL2 expression. Monocyte migration is significantly hampered in miR‐210‐3p mimic‐transfected HIF‐1A silenced cancer cells. In contrast, inhibition of miR‐210‐3p in HIF‐1A‐overexpressed cells markedly restored monocyte migration, highlighting a direct link between the miR‐210‐3p level and tumor monocyte burden. Moreover, miR‐210‐3p inhibition in 3D tumor spheroids promotes monocyte recruitment and skewing towards an antitumor M1 phenotype. Anti‐hsa‐miR‐210‐3p‐locked nucleic acid (LNA) delivery in a lung tumor xenograft zebrafish model caused tumor regression, suggesting that miR‐210‐3p could be a promising target for immunomodulatory therapeutic strategies against lung adenocarcinoma. Hypoxia‐associated stabilization of HIF‐1A markedly enhanced miR‐210‐3p expression, which directly targets CCL2 downregulation in lung adenocarcinoma, resulting in impaired monocyte infiltration and the stimulation of macrophage TAM polarization. Inhibition of miR‐210‐3p promoted monocyte recruitment and skewed towards an M1 phenotype in 3D tumor spheroids and a tumor xenograft model. Targeting miR‐210‐3p could be an effective therapeutic strategy for the management of lung adenocarcinoma.