DNA methylation at individual CpG-sites of EPB41L3, HTERT and FAM19A4 are useful for detection of cervical high-grade squamous intraepithelial lesions (HSIL) or worse: Analysis of individual CpG-sites outperforms averaging

Global methylation analysis of gene promoters is promising for detection of high-grade squamous intraepithelial lesions or worse (HSIL+) in high-risk human papillomavirus (hrHPV)-positive women. However, diagnostic performance of methylation data at individual CpG-sites is limited. We explored methy...

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Main Authors: Monica Molano, Dorothy A. Machalek, Samuel Phillips, Grace Tan, Suzanne M. Garland, David Hawkes, Prisha Balgovind, Reza Haqshenas, Steve G. Badman, John Bolnga, Josephine Gabuzzi, Zure Kombati, Gloria M. Munnull, Julia ML. Brotherton, Marion Saville, John M. Kaldor, Pamela J. Toliman, Andrew J. Vallely, Gerald L. Murray
Format: Article
Language:English
Published: Elsevier 2024-12-01
Series:Tumour Virus Research
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Online Access:http://www.sciencedirect.com/science/article/pii/S2666679024000120
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author Monica Molano
Dorothy A. Machalek
Samuel Phillips
Grace Tan
Suzanne M. Garland
David Hawkes
Prisha Balgovind
Reza Haqshenas
Steve G. Badman
John Bolnga
Josephine Gabuzzi
Zure Kombati
Gloria M. Munnull
Julia ML. Brotherton
Marion Saville
John M. Kaldor
Pamela J. Toliman
Andrew J. Vallely
Gerald L. Murray
author_facet Monica Molano
Dorothy A. Machalek
Samuel Phillips
Grace Tan
Suzanne M. Garland
David Hawkes
Prisha Balgovind
Reza Haqshenas
Steve G. Badman
John Bolnga
Josephine Gabuzzi
Zure Kombati
Gloria M. Munnull
Julia ML. Brotherton
Marion Saville
John M. Kaldor
Pamela J. Toliman
Andrew J. Vallely
Gerald L. Murray
author_sort Monica Molano
collection DOAJ
description Global methylation analysis of gene promoters is promising for detection of high-grade squamous intraepithelial lesions or worse (HSIL+) in high-risk human papillomavirus (hrHPV)-positive women. However, diagnostic performance of methylation data at individual CpG-sites is limited. We explored methylation for predicting HSIL+ in self- and clinician-collected samples from Papua New Guinea.Methylation of EPB41L3 (1–6 CpG-sites), hTERT (1–10 CpG-sites) and FAM19A4 (1–5 CpG-sites) was assessed through pyrosequencing from 44 HPV+ samples (4 cancers, 19 HSIL, 4 low-grade squamous intraepithelial lesions (LSIL), 17 normal). New primers were designed for FAM19A4 directed to the first exon region not explored previously.In clinician-collected samples, methylation at CpG-sites 4 and 5 of EPB41L3 were the best HSIL predictors (AUC >0.83) and CpG-site 4 for cancer (0.925). Combination of EPB41L3 sites 2/4 plus FAM19A4 site 1 were the best HSIL+ markers [100% sensitivity, 63.2% specificity].Methylation at CpG-site 5 of FAM19A4 was the best HSIL predictor (0.67) in self-collected samples, and CpG-sites 1 and 3 of FAM19A4 for cancer (0.77). Combined, FAM19A4 site 1 plus HPV 16/18 detection yielded sensitivity of 82.6% and specificity of 61.9%.In conclusion, methylation at individual CpG-sites of EPB41L3 and FAM19A4 outperformed global analysis and improved HSIL+ detection, warranting further investigation.
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spelling doaj-art-ac0c71d30e3949adb60b51b194c457ea2024-12-15T06:17:19ZengElsevierTumour Virus Research2666-67902024-12-0118200288DNA methylation at individual CpG-sites of EPB41L3, HTERT and FAM19A4 are useful for detection of cervical high-grade squamous intraepithelial lesions (HSIL) or worse: Analysis of individual CpG-sites outperforms averagingMonica Molano0Dorothy A. Machalek1Samuel Phillips2Grace Tan3Suzanne M. Garland4David Hawkes5Prisha Balgovind6Reza Haqshenas7Steve G. Badman8John Bolnga9Josephine Gabuzzi10Zure Kombati11Gloria M. Munnull12Julia ML. Brotherton13Marion Saville14John M. Kaldor15Pamela J. Toliman16Andrew J. Vallely17Gerald L. Murray18Centre for Women's Infectious Diseases, The Royal Women's Hospital, Melbourne, Victoria, Australia; Murdoch Children's Research Institute, Melbourne, Victoria, Australia; Corresponding author.Centre for Women's Infectious Diseases, The Royal Women's Hospital, Melbourne, Victoria, Australia; The Kirby Institute, University of New South Wales, Sydney, NSW, AustraliaCentre for Women's Infectious Diseases, The Royal Women's Hospital, Melbourne, Victoria, Australia; Murdoch Children's Research Institute, Melbourne, Victoria, AustraliaAustralian Centre for the Prevention of Cervical Cancer, Melbourne, Victoria, AustraliaCentre for Women's Infectious Diseases, The Royal Women's Hospital, Melbourne, Victoria, Australia; Murdoch Children's Research Institute, Melbourne, Victoria, Australia; Department of Obstetrics and Gynaecology, University of Melbourne, Parkville, Victoria, AustraliaAustralian Centre for the Prevention of Cervical Cancer, Melbourne, Victoria, AustraliaCentre for Women's Infectious Diseases, The Royal Women's Hospital, Melbourne, Victoria, Australia; Murdoch Children's Research Institute, Melbourne, Victoria, AustraliaCentre for Women's Infectious Diseases, The Royal Women's Hospital, Melbourne, Victoria, Australia; Murdoch Children's Research Institute, Melbourne, Victoria, Australia; Department of Obstetrics and Gynaecology, University of Melbourne, Parkville, Victoria, AustraliaThe Kirby Institute, University of New South Wales, Sydney, NSW, AustraliaPapua New Guinea Institute of Medical Research, Goroka, Papua New Guinea; Department of Obstetrics and Gynaecology, Modilon General Hospital, Madang, Papua New GuineaPapua New Guinea Institute of Medical Research, Goroka, Papua New Guinea; Department of Obstetrics and Gynaecology, Modilon General Hospital, Madang, Papua New GuineaDepartment of Pathology, Mt Hagen Provincial Hospital, WHP 281, Papua New GuineaDepartment of Obstetrics and Gynaecology, Mt Hagen Provincial Hospital, Mount Hagen, Papua New Guinea; Papua New Guinea Institute of Medical Research, Goroka, Papua New GuineaAustralian Centre for the Prevention of Cervical Cancer, Melbourne, Victoria, AustraliaAustralian Centre for the Prevention of Cervical Cancer, Melbourne, Victoria, AustraliaThe Kirby Institute, University of New South Wales, Sydney, NSW, AustraliaThe Kirby Institute, University of New South Wales, Sydney, NSW, AustraliaThe Kirby Institute, University of New South Wales, Sydney, NSW, AustraliaCentre for Women's Infectious Diseases, The Royal Women's Hospital, Melbourne, Victoria, Australia; Murdoch Children's Research Institute, Melbourne, Victoria, Australia; Department of Obstetrics and Gynaecology, University of Melbourne, Parkville, Victoria, AustraliaGlobal methylation analysis of gene promoters is promising for detection of high-grade squamous intraepithelial lesions or worse (HSIL+) in high-risk human papillomavirus (hrHPV)-positive women. However, diagnostic performance of methylation data at individual CpG-sites is limited. We explored methylation for predicting HSIL+ in self- and clinician-collected samples from Papua New Guinea.Methylation of EPB41L3 (1–6 CpG-sites), hTERT (1–10 CpG-sites) and FAM19A4 (1–5 CpG-sites) was assessed through pyrosequencing from 44 HPV+ samples (4 cancers, 19 HSIL, 4 low-grade squamous intraepithelial lesions (LSIL), 17 normal). New primers were designed for FAM19A4 directed to the first exon region not explored previously.In clinician-collected samples, methylation at CpG-sites 4 and 5 of EPB41L3 were the best HSIL predictors (AUC >0.83) and CpG-site 4 for cancer (0.925). Combination of EPB41L3 sites 2/4 plus FAM19A4 site 1 were the best HSIL+ markers [100% sensitivity, 63.2% specificity].Methylation at CpG-site 5 of FAM19A4 was the best HSIL predictor (0.67) in self-collected samples, and CpG-sites 1 and 3 of FAM19A4 for cancer (0.77). Combined, FAM19A4 site 1 plus HPV 16/18 detection yielded sensitivity of 82.6% and specificity of 61.9%.In conclusion, methylation at individual CpG-sites of EPB41L3 and FAM19A4 outperformed global analysis and improved HSIL+ detection, warranting further investigation.http://www.sciencedirect.com/science/article/pii/S2666679024000120DNA methylationCervical cancerHuman papillomavirusDiagnostic testEpigeneticsMolecular diagnostics
spellingShingle Monica Molano
Dorothy A. Machalek
Samuel Phillips
Grace Tan
Suzanne M. Garland
David Hawkes
Prisha Balgovind
Reza Haqshenas
Steve G. Badman
John Bolnga
Josephine Gabuzzi
Zure Kombati
Gloria M. Munnull
Julia ML. Brotherton
Marion Saville
John M. Kaldor
Pamela J. Toliman
Andrew J. Vallely
Gerald L. Murray
DNA methylation at individual CpG-sites of EPB41L3, HTERT and FAM19A4 are useful for detection of cervical high-grade squamous intraepithelial lesions (HSIL) or worse: Analysis of individual CpG-sites outperforms averaging
Tumour Virus Research
DNA methylation
Cervical cancer
Human papillomavirus
Diagnostic test
Epigenetics
Molecular diagnostics
title DNA methylation at individual CpG-sites of EPB41L3, HTERT and FAM19A4 are useful for detection of cervical high-grade squamous intraepithelial lesions (HSIL) or worse: Analysis of individual CpG-sites outperforms averaging
title_full DNA methylation at individual CpG-sites of EPB41L3, HTERT and FAM19A4 are useful for detection of cervical high-grade squamous intraepithelial lesions (HSIL) or worse: Analysis of individual CpG-sites outperforms averaging
title_fullStr DNA methylation at individual CpG-sites of EPB41L3, HTERT and FAM19A4 are useful for detection of cervical high-grade squamous intraepithelial lesions (HSIL) or worse: Analysis of individual CpG-sites outperforms averaging
title_full_unstemmed DNA methylation at individual CpG-sites of EPB41L3, HTERT and FAM19A4 are useful for detection of cervical high-grade squamous intraepithelial lesions (HSIL) or worse: Analysis of individual CpG-sites outperforms averaging
title_short DNA methylation at individual CpG-sites of EPB41L3, HTERT and FAM19A4 are useful for detection of cervical high-grade squamous intraepithelial lesions (HSIL) or worse: Analysis of individual CpG-sites outperforms averaging
title_sort dna methylation at individual cpg sites of epb41l3 htert and fam19a4 are useful for detection of cervical high grade squamous intraepithelial lesions hsil or worse analysis of individual cpg sites outperforms averaging
topic DNA methylation
Cervical cancer
Human papillomavirus
Diagnostic test
Epigenetics
Molecular diagnostics
url http://www.sciencedirect.com/science/article/pii/S2666679024000120
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