ADCY8 gene methylation in plasma as a predictive marker of breast cancer neoadjuvant chemotherapy

Background. Neoadjuvant chemotherapy (NAC) is a common practice for locally advanced breast cancer to downstage the disease to become operable. The top-of-the-line predictor of the effectiveness of neoadjuvant treatment is the pathologic complete response, defined as the absence of viable tumor cells in the mammary gland and regional lymph nodes. According to different studies, this result is achieved in no more than 13-33% of patients. Increasing the effectiveness of NACHT can be achieved through the identification of predictive markers that allow assessing the sensitivity to therapy. Objective. To assess the methylation status of SLC9A3, DPYS, IRF4, ADCY8, KCNQ2, TERT, SYNDIG1 and SKOR2 genes in tumor and serum samples and to analyze its association with response to breast cancer neoadjuvant chemotherapy. Material and methods. Core biopsy and plasma samples were obtained before and after neoadjuvant chemotherapy from 36 primary breast cancer patients. DNA methylation status was assessed using methylation sensitive PCR. Result. DNA methylation analysis of tumor biopsy material obtained before treatment showed the following gene methylation frequencies: SLC9A3 - 27.8% (10/36), DPYS - 8.3% (3/36), IRF4 - 22.2% (8/36), ADCY8 - 41.7% (15/36), KCNQ2 - 27.8% (10/36), TERT - 8.3% (3/36), SYNDIG1 - 16.7% (6/36) and SKOR2 - 5.5% (2/36). For further investigation in the blood plasma, 3 genes with the highest methylation frequencies were selected: SLC9A3 , KCNQ2 and ADCY8 . For the ADCY8 gene in the blood plasma, a statistically significant difference in methylation frequencies (p = 0.0076, exact two-sided Fisher test) was found between groups with different degrees of therapeutic response (methylation was more often observed in the group with poor response to treatment). Conclusion. Our results indicate that the methylation status of ADCY8 gene in plasma before treatment is associated with NAC pathological complete response in breast cancer.

рак молочной железы, неоадъювантная химиотерапия, маркеры метилирования, плазма крови, Breast cancer, neoadjuvant chemotherapy, methylation markers, blood plasma

1. Лазарева М.А., Павлов М.В., Шумская И.С., Овчинникова Е.Г., Ильинская О.Е., Орлова А.Г., Шахова Н.М., Масленникова А.В. Анализ эффективности неоадъювантной химиотерапии рака молочной железы. Онкогинекология. 2016. 4, 26-33.

2. Parekh T, Dodwell D, Sharma N, Shaaban A. Radiological and pathological predictors of response to neoadjuvant chemotherapy in breast cancer: a brief literature review. Pathobiology. 2015 82(3-4):124-32.

3. Fayanju OM, Ren Y, Thomas SM, Greenup RA, Plichta JK, Rosenberger L, Tamirisa N, Force J, Boughey JC, Hyslop T, Hwang ES. The Clinical Significance of Breast-only and Node-only Pathologic Complete Response (pCR) After Neoadjuvant Chemotherapy (NACT): A Review of 20,000 Breast Cancer Patients in the National Cancer Data Base (NCDB). Ann Surg. 2018. doi: 10.1097/SLA.0000000000002953.

4. Watanabe Y, Maeda I, Oikawa R, Wu W, Tsuchiya K, Miyoshi Y, Itoh F, Tsugawa K, Ohta T. Aberrant DNA methylation status of DNA repair genes in breast cancer treated with neoadjuvant chemotherapy. Genes Cells. 2013. 18(12):1120-30.

5. Tsang JS, Vencken S, Sharaf O, Leen E, Kay EW, McNamara DA, Deasy J, Mulligan ED. Global DNA methylation is altered by neoadjuvant chemoradiotherapy in rectal cancer and may predict response to treatment - A pilot study. Eur J Surg Oncol. 2014. 40(11):1459-66.

6. Takahashi H, Kagara N, Tanei T, Naoi Y, Shimoda M, Shimomura A, Shimazu K, Kim S, Noguchi S. Correlation of Methylated Circulating Tumor DNA With Response to Neoadjuvant Chemotherapy in Breast Cancer Patients. Clinical Breast Cancer. 2017. 17(1):61-69e3.

7. Bhangu J, Beer A, Mittlbock M, Tamandl D, Pulverer W, Taghizadeh H, Stremitzer S, Kaczirek K, Gruenberger T, Gnant M, Bergmann M, Mannhalter C, Weinhousel A, Oehler R, Bachleitner-Hofmann T. Circulating Free Methylated Tumor DNA Markers for Sensitive Assessment of Tumor Burden and Early Response Monitoring in Patients Receiving Systemic Chemotherapy for Colorectal Cancer Liver Metastasis. Ann Surg. 2018. doi: 10.1097/SLA.0000000000002901.

8. Гарбуков Е.Ю., Слонимская Е.М., Красулина Н.А., Дорошенко А.В., Кокорина Ю.Л. Неоадъювантная терапия при раке молочной железы. Сибирский онкологический журнал. 2005. 2 (14):63.

9. Tanas A.S., Poddubskaya E.V., Kekeeva T.V., Trotsenko I.D., Kuznetsova E.B., Zaletayev D.V., Strelnikov V.V. Breast cancer molecular classification based on DNA methylation assessed by reduced representation bisulfite sequencing. Eur J Hum Genet. 2015. 23 (S1):263.

10. Сигин В.О., Кузнецова Е.Б., Симонова О.А., Жевлова А.И., Литвяков Н.В., Слонимская Е.М., Цыганов М.М., Володин И.В., Шикеева А.А., Стрельников В.В., Залетаев Д.В., Танас А.С. Медицинская ДНК-технология оценки чувствительности опухолей молочной железы люминального В подтипа к неоадъювантной химиотерапии с применением антрациклинов на основе маркеров метилирования ДНК. Медицинская генетика. 2017;16(10):29-35.

11. Wang X, Xiao F, Li Q, Liu J, He Y,1 K. Qing-Peng. Large-scale DNA methylation expression analysis across 12 solid cancers reveals hypermethylation in the calcium-signaling pathway. Oncotarget. 2017. 8(7):11868-11876.

12. Shen-Gunther J, Wang CM, Poage GM, Lin CL, Perez L, Banks NA, Huang TH. Molecular Pap smear: HPV genotype and DNA methylation of ADCY8, CDH8, and ZNF582 as an integrated biomarker for high-grade cervical cytology. Clin Epigenetics. 2016. 8:96. doi: 10.1186/s13148-016-0263-9.