DNA Methylation Status of HYAL1 in Malignant and Benign Thyroid Nodules

Abstract

Differentiation between benign and malignant thyroid nodules has been a challenge in clinical practice. Exploring a novel biomarker to determine the malignancy of thyroid nodules has important implications. We semi-quantitatively determined the DNA methylation levels of four CpG sites located at the gene body of HYAL1 in formalin-fixed paraffin-embedded (FFPE) tissue samples from 190 early-stage papillary thyroid cancer (PTC) cases and 190 age- and gender-matched subjects with benign thyroid nodule (BTN). HYAL1 expression was evaluated by immunohistochemical (IHC) staining in another cohort of 55 PTC and 55 matched BTN cases. Covariates-adjusted odds ratios (ORs) for 10% increased methylation were calculated by binary logistic regression. A 165 bp amplicon covering four CpG sites at the second exon of HYAL1 gene was designed. After adjusted for all covariates, higher methylation level of HYAL1_CpG_3,4 in the FFPE tissue was associated with PTC (OR per 10% increased methylation=1.53, p=0.025), even with stage І PTC (OR per 10% increased methylation=1.58, p=0.021). Hypermethylation of HYAL1_CpG_3,4 had a significant association with early-stage PTC in the females (OR per 10% increased methylation=1.60, p=0.028) rather than in the males. Besides, we found the higher expression of HYAL1 protein in PTC than that in BTN patients (IHC score: 2.3 vs. 0.5, p=1.00E-06). Our study suggested altered methylation and expression of HYAL1 could be a novel and potential biomarker in distinguishing malignant and benign thyroid nodules.

Key words

HYAL1 - DNA methylation - expression - identification - thyroid nodules

Publikationsverlauf

Eingereicht: 26. Mai 2022

Angenommen nach Revision: 29. September 2023

Artikel online veröffentlicht:
01. Dezember 2023

• References

• 1 Sanabria A, Kowalski LP, Shah JP. et al. Growing incidence of thyroid carcinoma in recent years: Factors underlying overdiagnosis. Head Neck 2018; 40: 855-866
• 2 Bogovic Crncic T, Ilic Tomas M, Girotto N. et al. Risk factors for thyroid cancer: what do we know so far?. Acta Clin Croat 2020; 59: 66-72
• 3 Schlumberger M, Leboulleux S. Current practice in patients with differentiated thyroid cancer. Nat Rev Endocrinol 2021; 17: 176-188
• 4 Feldkamp J, Fuhrer D, Luster M. et al. Fine needle aspiration in the investigation of thyroid nodules. Dtsch Arztebl Int 2016; 113: 353-359
• 5 Rana C, Singh KR, Ramakant P. et al. Impact of cytological pitfalls in the Bethesda system of reporting thyroid cytopathology, on surgical decision-making of patients with thyroid nodules: can these pitfalls be avoided?. Cytopathology 2021; 32: 192-204
• 6 Cibas ES, Ali SZ. The 2017 Bethesda system for reporting thyroid cytopathology. Thyroid 2017; 27: 1341-1346
• 7 Stewart R, Leang YJ, Bhatt CR. et al. Quantifying the differences in surgical management of patients with definitive and indeterminate thyroid nodule cytology. Eur J Surg Oncol 2020; 46: 252-257
• 8 Eszlinger M, Hegedus L, Paschke R. Ruling in or ruling out thyroid malignancy by molecular diagnostics of thyroid nodules. Best Pract Res Clin Endocrinol Metab 2014; 28: 545-557
• 9 Silaghi CA, Lozovanu V, Georgescu CE. et al. Thyroseq v3, afirma GSC, and microRNA panels versus previous molecular tests in the preoperative diagnosis of indeterminate thyroid nodules: a systematic review and meta-analysis. Front Endocrinol (Lausanne) 2021; 12: 649522
• 10 Grani G, Sponziello M, Pecce V. et al. Contemporary thyroid nodule evaluation and management. J Clin Endocrinol Metab 2020; 105: 2869-2883
• 11 Greenberg MVC, Bourc’his D. The diverse roles of DNA methylation in mammalian development and disease. Nat Rev Mol Cell Biol 2019; 20: 590-607
• 12 Zafon C, Gil J, Perez-Gonzalez B. et al. DNA methylation in thyroid cancer. Endocr Relat Cancer 2019; 26: R415-R439
• 13 Csoka AB, Scherer SW, Stern R. Expression analysis of six paralogous human hyaluronidase genes clustered on chromosomes 3p21 and 7q31. Genomics 1999; 60: 356-361
• 14 Vigetti D, Viola M, Karousou E. et al. Epigenetics in extracellular matrix remodeling and hyaluronan metabolism. FEBS J 2014; 281: 4980-4992
• 15 Hautmann SH, Lokeshwar VB, Schroeder GL. et al. Elevated tissue expression of hyaluronic acid and hyaluronidase validates the HA-HAase urine test for bladder cancer. J Urol 2001; 165: 2068-2074
• 16 Lokeshwar VB, Rubinowicz D, Schroeder GL. et al. Stromal and epithelial expression of tumor markers hyaluronic acid and HYAL1 hyaluronidase in prostate cancer. J Biol Chem 2001; 276: 11922-11932
• 17 Franzmann EJ, Schroeder GL, Goodwin WJ. et al. Expression of tumor markers hyaluronic acid and hyaluronidase (HYAL1) in head and neck tumors. Int J Cancer 2003; 106: 438-445
• 18 Tan JX, Wang XY, Su XL. et al. Upregulation of HYAL1 expression in breast cancer promoted tumor cell proliferation, migration, invasion and angiogenesis. PLoS One 2011; 6: e22836
• 19 Tan JX, Wang XY, Li HY. et al. HYAL1 overexpression is correlated with the malignant behavior of human breast cancer. Int J Cancer 2011; 128: 1303-1315
• 20 McAtee CO, Berkebile AR, Elowsky CG. et al. Hyaluronidase hyal1 increases tumor cell proliferation and motility through accelerated vesicle trafficking. J Biol Chem 2015; 290: 13144-13156
• 21 Yin Y, Li H, Yang C. et al. Detection of DNA methylation of HYAL2 gene for differentiating malignant from benign thyroid tumors. Nan Fang Yi Ke Da Xue Xue Bao 2022; 42: 123-129
• 22 Yang R, Stocker S, Schott S. et al. The association between breast cancer and S100P methylation in peripheral blood by multicenter case-control studies. Carcinogenesis 2017; 38: 312-320
• 23 Kupers LK, Monnereau C, Sharp GC. et al. Meta-analysis of epigenome-wide association studies in neonates reveals widespread differential DNA methylation associated with birthweight. Nat Commun 2019; 10: 1893
• 24 Vehmeijer FOL, Kupers LK, Sharp GC. et al. DNA methylation and body mass index from birth to adolescence: meta-analyses of epigenome-wide association studies. Genome Med 2020; 12: 105
• 25 Ralhan R, Veyhl J, Chaker S. et al. Immunohistochemical subcellular localization of protein biomarkers distinguishes benign from malignant thyroid nodules: potential for fine-needle aspiration biopsy clinical application. Thyroid 2015; 25: 1224-1234
• 26 Huang H, Rusiecki J, Zhao N. et al. Thyroid-stimulating hormone, thyroid hormones, and risk of papillary thyroid cancer: a nested case-control study. Cancer Epidemiol Biomarkers Prev 2017; 26: 1209-1218
• 27 Han R, Sun W, Huang J. et al. Sex-biased DNA methylation in papillary thyroid cancer. Biomark Med 2021; 15: 109-120
• 28 Teschendorff AE, West J, Beck S. Age-associated epigenetic drift: implications, and a case of epigenetic thrift?. Hum Mol Genet 2013; 22: R7-R15
• 29 Wong R, Farrell SG, Grossmann M. Thyroid nodules: diagnosis and management. Med J Aust 2018; 209: 92-98
• 30 Locke WJ, Guanzon D, Ma C. et al. DNA Methylation Cancer Biomarkers: Translation to the Clinic. Front Genet 2019; 10: 1150
• 31 Du L, Zhao Z, Zheng R. et al. Epidemiology of thyroid cancer: incidence and mortality in China, 2015. Front Oncol 2020; 10: 1702
• 32 Mannathazhathu AS, George PS, Sudhakaran S. et al. Reproductive factors and thyroid cancer risk: Meta-analysis. Head Neck 2019; 41: 4199-4208
• 33 Caini S, Gibelli B, Palli D. et al. Menstrual and reproductive history and use of exogenous sex hormones and risk of thyroid cancer among women: a meta-analysis of prospective studies. Cancer Causes Control 2015; 26: 511-518
• 34 Liu J, Xu T, Ma L. et al. Signal pathway of estrogen and estrogen receptor in the development of thyroid cancer. Front Oncol 2021; 11: 593479
• 35 Gajowiec A, Chromik A, Furga K. et al. Is male sex a prognostic factor in papillary thyroid cancer?. J Clin Med 2021; 10: 2438
• 36 Ness C, Katta K, Garred O. et al. Integrated differential DNA methylation and gene expression of formalin-fixed paraffin-embedded uveal melanoma specimens identifies genes associated with early metastasis and poor prognosis. Exp Eye Res 2021; 203: 108426
• 37 Pelch KE, Tokar EJ, Merrick BA. et al. Differential DNA methylation profile of key genes in malignant prostate epithelial cells transformed by inorganic arsenic or cadmium. Toxicol Appl Pharmacol 2015; 286: 159-167
• 38 da Costa Prando E, Cavalli LR, Rainho CA. Evidence of epigenetic regulation of the tumor suppressor gene cluster flanking RASSF1 in breast cancer cell lines. Epigenetics 2011; 6: 1413-1424
• 39 Lokeshwar VB, Cerwinka WH, Isoyama T. et al. HYAL1 hyaluronidase in prostate cancer: a tumor promoter and suppressor. Cancer Res 2005; 65: 7782-7789
• 40 Kobayashi T, Chanmee T, Itano N. Hyaluronan: metabolism and function. Biomolecules 2020; 10: 1525
• 41 Jin Z, Zhang G, Liu Y. et al. The suppressive role of HYAL1 and HYAL2 in the metastasis of colorectal cancer. J Gastroenterol Hepatol 2019; 34: 1766-1776
• 42 Kohi S, Sato N, Cheng XB. et al. Increased expression of HYAL1 in pancreatic ductal adenocarcinoma. Pancreas 2016; 45: 1467-1473
• 43 Lokeshwar VB, Lokeshwar BL, Pham HT. et al. Association of elevated levels of hyaluronidase, a matrix-degrading enzyme, with prostate cancer progression. Cancer Res 1996; 56: 651-657
• 44 Hamester F, Sturken C, Legler K. et al. Key role of hyaluronan metabolism for the development of brain metastases in triple-negative breast cancer. Cells 2022; 11: 3275
• 45 Kumar K, Kanojia D, Bentrem DJ. et al. Targeting BET proteins decreases hyaluronidase-1 in pancreatic cancer. Cells 2023; 12: 1490
• 46 Neri F, Rapelli S, Krepelova A. et al. Intragenic DNA methylation prevents spurious transcription initiation. Nature 2017; 543: 72-77
• 47 Jjingo D, Conley AB, Yi SV. et al. On the presence and role of human gene-body DNA methylation. Oncotarget 2012; 3: 462-474
• 48 Arechederra M, Bazai SK, Abdouni A. et al. ADAMTSL5 is an epigenetically activated gene underlying tumorigenesis and drug resistance in hepatocellular carcinoma. J Hepatol 2021; 74: 893-906
• 49 Yang X, Han H, De Carvalho DD. et al. Gene body methylation can alter gene expression and is a therapeutic target in cancer. Cancer Cell 2014; 26: 577-590
• 50 Moran S, Arribas C, Esteller M. Validation of a DNA methylation microarray for 850,000 CpG sites of the human genome enriched in enhancer sequences. Epigenomics 2016; 8: 389-399
• 51 Kling T, Wenger A, Beck S. et al. Validation of the methylationEPIC beadchip for fresh-frozen and formalin-fixed paraffin-embedded tumours. Clin Epigenetics 2017; 9: 33