BACKGROUND/AIM: Rectal cancer accounts for approximately one-third of all colorectal cancers. Currently, the standard treatment for locally advanced rectal cancer (LARC) is neoadjuvant chemoradiotherapy (CRT) with capecitabine or 5-fluorouracil followed by curative surgery. Unfortunately, only 20% of patients with LARC present complete pathological response after CRT, whereas in 20-40% cases the response is poor or absent. The aim of our study was to evaluate whether microRNAs (miRNAs) in tumor biopsy specimen have the potential to predict therapeutic response in LARC patients. PATIENTS AND METHODS: In total 87 LARC patients treated by CRT were enrolled in our prospective study. To identify predictive miRNAs, we used small RNA sequencing in 40 tumor biopsy samples of LARC patients (20 responders, 20 non-responders) and qPCR validation of selected miRNA candidates. RESULTS: In the discovery phase of the study, we identified 69 miRNAs to have significantly different expression between the group of responders (TRG 1,2) and a group of non-responders (TRG 4,5) to neoadjuvant CRT. Among these miRNAs, 48 showed a lower expression and 21 showed higher expression in tumor tissues from poorly responding LARC patients. Five miRNAs were selected for validation, but only miR-487a-3p was confirmed to have a significantly higher expression in the tumor biopsy specimens of non-responders to neoadjuvant CRT (p<0.0006, AUC=0.766). Gene Ontology (GO) clustering and pathway enrichment analysis of the miR-487a-3p mRNA targets, revealed potential mechanisms behind miR-487a-3p roles in chemoradioresistance (e.g. TGF-beta signaling pathway, protein kinase activity, double-stranded DNA binding, or microRNAs in cancer). CONCLUSION: By combination of miRNA expression profiling and integrative computational biology we identified miR-487a-3p as a potential predictive biomarker of CRT response in LARC patients.

Central European Institute of Technology Masaryk University Brno Czech Republic

Department of Comprehensive Cancer Care Masaryk Memorial Cancer Institute Brno Czech Republic

Department of Pathology Faculty Hospital Brno and Faculty of Medicine Masaryk University Brno Czech Republic

Department of Surgery 1 Faculty of Medicine and Dentistry Palacky University Olomouc and University Hospital Olomouc Olomouc Czech Republic

Department of Surgery Faculty Hospital Brno and Faculty of Medicine Masaryk University Brno Czech Republic

Department of Surgery Hospital and Oncological Centre Novy Jicin Novy Jicin Czech Republic

Department of Surgical Oncology Masaryk Memorial Cancer Institute Brno Czech Republic

MEDFUTURE Research Center for Advanced Medicine University of Medicine and Pharmacy Iuliu Hatieganu Cluj Napoca Romania

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Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2018;68:394–424. PMID: 30207593. DOI: 10.3322/caac.21492. PubMed

Iskander O, Courtot L, Tabchouri N, Artus A, Michot N, Muller O, Pabst-Giger U, Bourlier P, Kraemer-Bucur A, Lecomte T, Guyetant S, Chapet S, Calais G, Salamé E, Ouaïssi M. Complete pathological response following radiochemotherapy for locally advanced rectal cancer: Short and long-term outcome. Anticancer Res. 2019;39:5105–5113. PMID: 31519622. DOI: 10.21873/anticanres.13705. PubMed

Kim SH, Chang HJ, Kim DY, Park JW, Baek JY, Kim SY, Park SC, Oh JH, Yu A, Nam B-H. What is the ideal tumor regression grading system in rectal cancer patients after preoperative chemoradiotherapy. Cancer Res Treat. 2016;48:998–1009. PMID: 26511803. DOI: 10.4143/crt.2015.254. PubMed PMC

Mandard AM, Dalibard F, Mandard JC, Marnay J, Henry-Amar M, Petiot JF, Roussel A, Jacob JH, Segol P, Samama G. Pathologic assessment of tumor regression after preoperative chemoradiotherapy of esophageal carcinoma. Clinicopathologic correlations. Cancer. 1994;73:2680–2686. PMID: 8194005. DOI: 10.1002/1097-0142(19940601)73:11<2680::aid-cncr2820731105>3.0.co;2-c. PubMed

Du B, Wang X, Wu D, Wang T, Yang X, Wang J, Shi X, Chen L, Zhang W. MicroRNA expression profiles identify biomarkers for predicting the response to chemoradiotherapy in rectal cancer. Mol Med Rep. 2018 PMID: 29956755. DOI:10.3892/mmr.2018.9215. PubMed PMC

Slattery ML, Herrick JS, Mullany LE, Samowitz WS, Sevens JR, Sakoda L, Wolff RK. The co-regulatory networks of tumor suppressor genes, oncogenes, and miRNAs in colorectal cancer. Genes Chromosomes Cancer. 2017;56:769–787. PMID: 28675510. DOI: 10.1002/gcc.22481. PubMed PMC

Zhang L, Zhang Y, Wong SH, Law PTY, Zhao S, Yu J, Chan MTV, Wu WKK. Common deregulation of seven biological processes by MicroRNAs in gastrointestinal cancers. Sci Rep. 2018;8:3287. PMID: 29459716. DOI: 10.1038/s41598-018-21573-w. PubMed PMC

Wu W. Identification of aberrantly expressed miRNAs in rectal cancer. Oncol Rep. 2012 PMID: 22576798. DOI: 10.3892/or.2012.1769. PubMed

Gaedcke J, Grade M, Camps J, Sokilde R, Kaczkowski B, Schetter AJ, Difilippantonio MJ, Harris CC, Ghadimi BM, Moller S, Beissbarth T, Ried T, Litman T. The rectal cancer microRNAome - microRNA expression in rectal cancer and matched normal mucosa. Clin Cancer Res. 2012;18:4919–4930. PMID: 22850566. DOI: 10.1158/1078-0432.CCR-12-0016. PubMed PMC

Okugawa Y, Shirai Y, Nodono H, Matsutani F, Itoh M, Hishida A, Morimoto Y, Nishikawa R, Yokoe T, Tanaka K, Urata H, Toiyama Y, Araki T, Inoue Y, Tanaka M, Kusunoki M, Miki C. Objective predictive score as a feasible biomarker for short-term survival in terminalIy Ill patients with cancer. Anticancer Res. 2017;37:267–275. PMID: 28011502. DOI: 10.21873/anticanres.11317. PubMed

Orosz E, Kiss I, Gyöngyi Z, Varjas T. Expression of circulating miR-155, miR-21, miR-221, miR-30a, miR-34a and miR-29a: Comparison of colonic and rectal cancer. In Vivo. 2018;32:1333–1337. PMID: 30348685. DOI: 10.21873/invivo.11383. PubMed PMC

Bullock MD, Pickard KM, Nielsen BS, Sayan AE, Jenei V, Mellone M, Mitter R, Primrose JN, Thomas GJ, Packham GK, Mirnezami AH. Pleiotropic actions of miR-21 highlight the critical role of deregulated stromal microRNAs during colorectal cancer progression. Cell Death Dis. 2013;4:e684–e684. PMID: 23788041. DOI: 10.1038/cddis.2013.213. PubMed PMC

Hotchi M. microRNA expression is able to predict response to chemoradiotherapy in rectal cancer. Mol Clin Oncol. 2012 PMID: 24649136. DOI: 10.3892/mco.2012.9. PubMed PMC

Svoboda M, Sana J, Fabian P, Kocakova I, Gombosova J, Nekvindova J, Radova L, Vyzula R, Slaby O. MicroRNA expression profile associated with response to neoadjuvant chemoradiotherapy in locally advanced rectal cancer patients. Radiat Oncol. 2012;7 PMID: 23167930. DOI: 10.1186/1748-717X-7-195. PubMed PMC

Kheirelseid EAH, Miller N, Chang KH, Curran C, Hennessey E, Sheehan M, Newell J, Lemetre C, Balls G, Kerin MJ. miRNA expressions in rectal cancer as predictors of response to neoadjuvant chemoradiation therapy. Int J Colorectal Dis. 2013;28:247–260. PMID: 22903298. DOI: 10.1007/s00384-012-1549-9. PubMed

Lopes-Ramos CM, Habr-Gama A, Quevedo B de S, Felício NM, Bettoni F, Koyama FC, Asprino PF, Galante PA, Gama-Rodrigues J, Camargo AA, Perez RO, Parmigiani RB. Overexpression of miR-21-5p as a predictive marker for complete tumor regression to neoadjuvant chemoradiotherapy in rectal cancer patients. BMC Med Genomics. 2014;7 PMID: 25496125. DOI: 10.1186/s12920-014-0068-7. PubMed PMC

Caramés C, Cristobal I, Moreno V, Marín J, González-Alonso P, Torrejón B, Minguez P, Leon A, Martín J, Hernández R, Pedregal M, Martín M, Cortés D, García-Olmo D, Fernández M, Rojo F, García-Foncillas J. MicroRNA-31 emerges as a predictive biomarker of pathological response and outcome in locally advanced rectal cancer. Int J Mol Sci. 2016;17:878. PMID: 27271609. DOI: 10.3390/ijms17060878. PubMed PMC

Salvi S, Molinari C, Foca F, Teodorani N, Saragoni L, Puccetti M, Passardi A, Tamberi S, Avanzolini A, Lucci E, Calistri D. miR-17-92a-1 cluster host gene (MIR17HG) evaluation and response to neoadjuvant chemoradiotherapy in rectal cancer. OncoTargets Ther. 2016;9:2735–2742. PMID: 27226732. DOI: 10.2147/OTT.S105760. PubMed PMC

Millino C, Maretto I, Pacchioni B, Digito M, De Paoli A, Canzonieri V, D’Angelo E, Agostini M, Rizzolio F, Giordano A, Barina A, Rajendran S, Esposito G, Lanfranchi G, Nitti D, Pucciarelli S. Gene and MicroRNA expression are predictive of tumor response in rectal adenocarcinoma patients treated with preoperative chemoradiotherapy: Combined miRNAS and gene expression. J Cell Physiol. 2017;232:426–435. PMID: 27225591. DOI: 10.1002/jcp.25441. PubMed

Eriksen AHM, Sørensen FB, Andersen RF, Jakobsen A, Hansen TF. Association between the expression of microRNAs and the response of patients with locally advanced rectal cancer to preoperative chemoradiotherapy. Oncol Lett. 2017;14:201–209. PMID: 28693154. DOI: 10.3892/ol.2017.6141. PubMed PMC

D’Angelo E, Zanon C, Sensi F, Digito M, Rugge M, Fassan M, Scarpa M, Pucciarelli S, Nitti D, Agostini M. miR-194 as predictive biomarker of responsiveness to neoadjuvant chemoradiotherapy in patients with locally advanced rectal adenocarcinoma. J Clin Pathol. 2018;71:344–350. PMID: 28870889. DOI: 10.1136/jclinpath-2017-204690. PubMed

Campayo M, Navarro A, Benítez JC, Santasusagna S, Ferrer C, Monzó M, Cirera L. miR-21, miR-99b and miR-375 combination as predictive response signature for preoperative chemoradiotherapy in rectal cancer. PLoS One. 2018;13:e0206542. PMID: 30388154. DOI: 10.1371/journal.pone.0206542. PubMed PMC

Wang M, Yu W, Gao J, Ma W, Frentsch M, Thiel A, Liu M, Rahman N, Qin Z, Li X. MicroRNA-487a-3p functions as a new tumor suppressor in prostate cancer by targeting CCND1. J Cell Physiol. 2020;235:1588–1600. PMID: 31309555. DOI: 10.1002/jcp.29078. PubMed

González-Vallinas M, Rodríguez-Paredes M, Albrecht M, Sticht C, Stichel D, Gutekunst J, Pitea A, Sass S, Sánchez-Rivera FJ, Lorenzo-Bermejo J, Schmitt J, De La Torre C, Warth A, Theis FJ, Müller NS, Gretz N, Muley T, Meister M, Tschaharganeh DF, Schirmacher P, Matthäus F, Breuhahn K. Epigenetically regulated chromosome 14q32 miRNA cluster induces metastasis and predicts poor prognosis in lung adenocarcinoma patients. Mol Cancer Res. 2018;16:390–402. PMID: 29330288. DOI: 10.1158/1541-7786.MCR-17-0334. PubMed

Zurawek M, Dzikiewicz-Krawczyk A, Izykowska K, Ziolkowska-Suchanek I, Skowronska B, Czainska M, Podralska M, Fichna P, Przybylski G, Fichna M, Nowak J. miR-487a-3p upregulated in type 1 diabetes targets CTLA4 and FOXO3. Diabetes Res Clin Pract. 2018;142:146–153. PMID: 29859273. DOI: 10.1016/j.diabres.2018.05.044. PubMed

Yang X, Wang M, Lin B, Yao D, Li J, Tang X, Li S, Liu Y, Xie R, Yu S. miR-487a promotes progression of gastric cancer by targeting TIA1. Biochimie. 2018;154:119–126. PMID: 30144499. DOI: 10.1016/j.biochi.2018.08.006. PubMed

Chang R-M, Xiao S, Lei X, Yang H, Fang F, Yang L-Y. miRNA-487a promotes proliferation and metastasis in hepatocellular carcinoma. Clin Cancer Res. 2017;23:2593–2604. PMID: 27827315. DOI: 10.1158/1078-0432.CCR-16-0851. PubMed

Ma M, He M, Jiang Q, Yan Y, Guan S, Zhang J, Yu Z, Chen Q, Sun M, Yao W, Zhao H, Jin F, Wei M. MiR-487a promotes TGF-β1-induced EMT, the migration and invasion of breast cancer cells by directly targeting MAGI2. Int J Biol Sci. 2016;12:397–408. PMID: 27019625. DOI: 10.7150/ijbs.13475. PubMed PMC

Skarkova V, Kralova V, Vitovcova B, Rudolf E. Selected aspects of chemoresistance mechanisms in colorectal carcinoma – A focus on epithelial-to-mesenchymal transition, autophagy, and apoptosis. Cells. 2019;8:234. PMID: 30871055. DOI: 10.3390/cells8030234. PubMed PMC

Ieda T, Tazawa H, Okabayashi H, Yano S, Shigeyasu K, Kuroda S, Ohara T, Noma K, Kishimoto H, Nishizaki M, Kagawa S, Shirakawa Y, Saitou T, Imamura T, Fujiwara T. Visualization of epithelial-mesenchymal transition in an inflammatory microenvironment–colorectal cancer network. Sci Rep. 2019;9:16378. PMID: 31705021. DOI: 10.1038/s41598-019-52816-z. PubMed PMC