Epigenetic Markers for Non-invasive Early Detection of Nasopharyngeal Carcinoma by Methylation-Sensitive High Resolution Melting
Xuesong Yang1, Wei Dai1, Dora Lai-wan Kwong1,2†, Carol Y.Y. Szeto1, Elibe Hiu-wun Wong1, Wai Tong Ng2,3, Anne W.M. Lee2,4, Roger K.C. Ngan2,5, Chun Chung Yau2,6, Stewart Y. Tung2,7, Maria Li Lung1,2†
1
Department of Clinical Oncology and Center for Cancer Research, University of
Hong Kong, Pokfulam, Hong Kong Special Administrative Region, People’s Republic of China 2
Center for Nasopharyngeal Carcinoma Research, University of Hong Kong, Hong
Kong Special Administrative Region, People’s Republic of China 3
Department of Clinical Oncology, Pamela Youde Nethersole Hospital, Hong Kong
Special Administrative Region, People’s Republic of China 4
Department of Clinical Oncology, University of Hong Kong-Shenzhen Hospital,
Shenzhen, People’s Republic of China 5
Department of Clinical Oncology, Queen Elizabeth Hospital, Hong Kong Special
Administrative Region, People’s Republic of China 6
Department of Oncology, Princess Margaret Hospital, Hong Kong Special
Administrative Region, People’s Republic of China 7
Department of Clinical Oncology, Tuen Mun Hospital, Hong Kong Special
Administrative Region, People’s Republic of China
This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process which may lead to differences between this version and the Version of Record. Please cite this article as an ‘Accepted Article’, doi: 10.1002/ijc.29192
International Journal of Cancer
Short Title: Non-invasive Early Detection of Nasopharyngeal Carcinoma
†
Correspondence: Maria Li Lung
Tel: (852) 3917-9783 Fax: (852) 2819-5872 E-mail: [email protected]
Address: Room L6-43, 6/F, Laboratory Block, Department of Clinical Oncology, University of Hong Kong, 21 Sassoon Road, Pokfulam, Hong Kong, HKSAR, People’s Republic of China
Dora L. Kwong Tel: (852) 22554521 Fax: (852) 28726426 E-mail: [email protected].
Address: Department of Clinical Oncology, University of Hong Kong, 1/F, Professorial Block, Queen Mary Hospital, Pokfulam, Hong Kong. HKSAR, People’s Republic of China
Keywords: Biomarker, DNA Methylation, NPC, MS-HRM, Early diagnosis
Abbreviations: NPC: Nasopharyngeal carcinoma; EBV: Epstein-Barr virus; TSG: tumor suppressor gene, MS-HRM: Methylation-sensitive high resolution melting; RASSF1A: Ras Association domain Family 1A; WIF1: Wnt inhibitory factor 1, DAPK1: Death-associated protein kinase 1; RARβ2: Retinoic acid receptor β2; PPV: positive predictive value; AJCC: American Joint Committee on Cancer; ROC: 2
John Wiley & Sons, Inc.
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International Journal of Cancer
Receiver operating characteristic; AUC: Area under curve.
Article Category: Research Articles- Early Detection and Diagnosis Brief Description: This study provide valuable information for use of DNA methylation biomarkers to supplement current methods in the screening of early NPC and the monitoring of local recurrence, using blood and nasopharyngeal (NP) brushings. The identification of appropriate biomarkers to improve screening of NPC using non-invasive screening procedures will facilitate identifying early NPC in suspected high-risk individuals to permit more timely and effective treatments, and more accurately monitoring post-treatment recurrence of NPC.
Abstract Nasopharyngeal carcinoma (NPC) is a human malignancy that is closely associated with Epstein-Barr Virus (EBV). Early diagnosis of NPC will greatly improve the overall survival. However, current EBV DNA marker detection still lacks the predictive value to perform well in high-risk populations for early detection of NPC. Since aberrant promoter hypermethylation of tumor suppressor genes (TSGs) is widely considered to be an important epigenetic change in early carcinogenesis, this study identified a panel of methylation markers for early detection of NPC and also assessed the clinical usefulness of these markers with non-invasive plasma specimens instead of biopsies. MS-HRM assays were carried out to assess the methylation status 3
John Wiley & Sons, Inc.
International Journal of Cancer
of a selected panel of four TSGs (RASSF1A, WIF1, DAPK1, RARβ2) in biopsies, NP brushings and cell-free plasma from NPC patients. High-risk and cancer-free groups were used as controls. DNA methylation panel showed higher sensitivity and specificity than EBV DNA marker in cell-free plasma from NPC patients at early stages (I-II), and in addition to the EBV DNA marker, MS-HRM test for plasma and NP brushing DNA methylation significantly increased the detection rate at all NPC stages as well as local recurrence, using this selected four-gene panel (p
Xuesong Yang1, Wei Dai1, Dora Lai-wan Kwong1,2†, Carol Y.Y. Szeto1, Elibe Hiu-wun Wong1, Wai Tong Ng2,3, Anne W.M. Lee2,4, Roger K.C. Ngan2,5, Chun Chung Yau2,6, Stewart Y. Tung2,7, Maria Li Lung1,2†
1
Department of Clinical Oncology and Center for Cancer Research, University of
Hong Kong, Pokfulam, Hong Kong Special Administrative Region, People’s Republic of China 2
Center for Nasopharyngeal Carcinoma Research, University of Hong Kong, Hong
Kong Special Administrative Region, People’s Republic of China 3
Department of Clinical Oncology, Pamela Youde Nethersole Hospital, Hong Kong
Special Administrative Region, People’s Republic of China 4
Department of Clinical Oncology, University of Hong Kong-Shenzhen Hospital,
Shenzhen, People’s Republic of China 5
Department of Clinical Oncology, Queen Elizabeth Hospital, Hong Kong Special
Administrative Region, People’s Republic of China 6
Department of Oncology, Princess Margaret Hospital, Hong Kong Special
Administrative Region, People’s Republic of China 7
Department of Clinical Oncology, Tuen Mun Hospital, Hong Kong Special
Administrative Region, People’s Republic of China
This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process which may lead to differences between this version and the Version of Record. Please cite this article as an ‘Accepted Article’, doi: 10.1002/ijc.29192
International Journal of Cancer
Short Title: Non-invasive Early Detection of Nasopharyngeal Carcinoma
†
Correspondence: Maria Li Lung
Tel: (852) 3917-9783 Fax: (852) 2819-5872 E-mail: [email protected]
Address: Room L6-43, 6/F, Laboratory Block, Department of Clinical Oncology, University of Hong Kong, 21 Sassoon Road, Pokfulam, Hong Kong, HKSAR, People’s Republic of China
Dora L. Kwong Tel: (852) 22554521 Fax: (852) 28726426 E-mail: [email protected].
Address: Department of Clinical Oncology, University of Hong Kong, 1/F, Professorial Block, Queen Mary Hospital, Pokfulam, Hong Kong. HKSAR, People’s Republic of China
Keywords: Biomarker, DNA Methylation, NPC, MS-HRM, Early diagnosis
Abbreviations: NPC: Nasopharyngeal carcinoma; EBV: Epstein-Barr virus; TSG: tumor suppressor gene, MS-HRM: Methylation-sensitive high resolution melting; RASSF1A: Ras Association domain Family 1A; WIF1: Wnt inhibitory factor 1, DAPK1: Death-associated protein kinase 1; RARβ2: Retinoic acid receptor β2; PPV: positive predictive value; AJCC: American Joint Committee on Cancer; ROC: 2
John Wiley & Sons, Inc.
Page 2 of 29
Page 3 of 29
International Journal of Cancer
Receiver operating characteristic; AUC: Area under curve.
Article Category: Research Articles- Early Detection and Diagnosis Brief Description: This study provide valuable information for use of DNA methylation biomarkers to supplement current methods in the screening of early NPC and the monitoring of local recurrence, using blood and nasopharyngeal (NP) brushings. The identification of appropriate biomarkers to improve screening of NPC using non-invasive screening procedures will facilitate identifying early NPC in suspected high-risk individuals to permit more timely and effective treatments, and more accurately monitoring post-treatment recurrence of NPC.
Abstract Nasopharyngeal carcinoma (NPC) is a human malignancy that is closely associated with Epstein-Barr Virus (EBV). Early diagnosis of NPC will greatly improve the overall survival. However, current EBV DNA marker detection still lacks the predictive value to perform well in high-risk populations for early detection of NPC. Since aberrant promoter hypermethylation of tumor suppressor genes (TSGs) is widely considered to be an important epigenetic change in early carcinogenesis, this study identified a panel of methylation markers for early detection of NPC and also assessed the clinical usefulness of these markers with non-invasive plasma specimens instead of biopsies. MS-HRM assays were carried out to assess the methylation status 3
John Wiley & Sons, Inc.
International Journal of Cancer
of a selected panel of four TSGs (RASSF1A, WIF1, DAPK1, RARβ2) in biopsies, NP brushings and cell-free plasma from NPC patients. High-risk and cancer-free groups were used as controls. DNA methylation panel showed higher sensitivity and specificity than EBV DNA marker in cell-free plasma from NPC patients at early stages (I-II), and in addition to the EBV DNA marker, MS-HRM test for plasma and NP brushing DNA methylation significantly increased the detection rate at all NPC stages as well as local recurrence, using this selected four-gene panel (p
