title: Laser capture microdissection as a tool to evaluate human papillomavirus genotyping and methylation as biomarkers of persistence and progression of anal lesions.
abstract: BACKGROUND
  Anal squamous cell carcinoma is preceded by persistent infection with high-risk human papillomavirus (HPV) and the cancer precursor, high-grade squamous intraepithelial lesion (HSIL). Detection of specific HPV genotypes and HPV-related biomarkers may be an option for primary anal screening. However, more data on the natural history of HPV-related anal lesions are required. The outcomes from this study will enhance our understanding of the clinical and biological behaviour of HPV-related anal lesions and inform the development of future HPV genotype and/or biomarker screening tests.
  
  
  METHODS
  HIV-negative and HIV-positive men who have sex with men, aged 35 years and over, recruited from community-based settings in Sydney, Australia, attend 6 clinic visits over 3 years. At the first 5 visits, participants undergo a digital anorectal examination, an anal swab for HPV genotyping and anal cytology, and high-resolution anoscopy with directed biopsy of any visible abnormalities that are suggestive of any abnormality suspicious of SIL. Tissue sections from participants diagnosed with histologically confirmed HSIL at the baseline clinic visit will undergo laser capture microdissection, HPV detection and genotyping, and quantitation of CpG methylation in baseline and follow-up biopsies. Histological and cytological findings in combination with HPV genotyping data will be used to identify persistent HSIL. HSIL will be stratified as non-persistent and persistent based on their status at 12 months. The performance of HPV genotype and methylation status in predicting disease persistence at 12 months will be assessed, along with associations with HIV status and other covariates such as age.
  
  
  BACKGROUND
  The St Vincent's Hospital Ethics Committee granted ethics approval for the study. Written informed consent is obtained from all individuals before any study-specific procedures are performed. Findings from this study will be disseminated to participants and the community through study newsletters, and through peer-reviewed publications and international conferences.
authors: Regional HPV Labnet Reference Laboratory, Department of Microbiology and Infectious Diseases, The Royal Women's Hospital, Parkville, Victoria, Australia Murdoch Children's Research Institute, Parkville, Victoria, Australia.
authors: Alyssa M
authors: AM
authors: Cornall
authors: Douglass Hanly Moir Pathology, Sydney, New South Wales, Australia.
authors: Jennifer M
authors: JM
authors: Roberts
authors: Regional HPV Labnet Reference Laboratory, Department of Microbiology and Infectious Diseases, The Royal Women's Hospital, Parkville, Victoria, Australia.
authors: Monica
authors: M
authors: Molano
authors: Regional HPV Labnet Reference Laboratory, Department of Microbiology and Infectious Diseases, The Royal Women's Hospital, Parkville, Victoria, Australia Murdoch Children's Research Institute, Parkville, Victoria, Australia.
authors: Dorothy A
authors: DA
authors: Machalek
authors: Regional HPV Labnet Reference Laboratory, Department of Microbiology and Infectious Diseases, The Royal Women's Hospital, Parkville, Victoria, Australia Murdoch Children's Research Institute, Parkville, Victoria, Australia.
authors: Samuel
authors: S
authors: Phillips
authors: Western Sydney Sexual Health Centre, University of Sydney, Sydney, New South Wales, Australia.
authors: Richard J
authors: RJ
authors: Hillman
authors: The Kirby Institute, UNSW Australia, Kensington, New South Wales, Australia.
authors: Andrew E
authors: AE
authors: Grulich
authors: The Kirby Institute, UNSW Australia, Kensington, New South Wales, Australia.
authors: Fengyi
authors: F
authors: Jin
authors: The Kirby Institute, UNSW Australia, Kensington, New South Wales, Australia.
authors: I Mary
authors: IM
authors: Poynten
authors: The Kirby Institute, UNSW Australia, Kensington, New South Wales, Australia RPA Sexual Health, Sydney Local Health District, Camperdown, New South Wales, Australia.
authors: David J
authors: DJ
authors: Templeton
authors: Regional HPV Labnet Reference Laboratory, Department of Microbiology and Infectious Diseases, The Royal Women's Hospital, Parkville, Victoria, Australia Department of Obstetrics and Gynaecology, University of Melbourne.
authors: Suzanne M
authors: SM
authors: Garland
authors: Regional HPV Labnet Reference Laboratory, Department of Microbiology and Infectious Diseases, The Royal Women's Hospital, Parkville, Victoria, Australia Department of Obstetrics and Gynaecology, University of Melbourne.
authors: Sepehr N
authors: SN
authors: Tabrizi
mesh_terms: D000328:Adult; D001003:Anal Canal / Q000473:pathology / Q000821:virology; D001005:Anus Neoplasms / Q000175:diagnosis*; D001315:Australia; D015415:Biomarkers / Q000032:analysis*; D002294:Carcinoma, Squamous Cell / Q000175:diagnosis / Q000473:pathology; D019175:DNA Methylation; D018450:Disease Progression; D005838:Genotype; D006679:HIV Seropositivity / Q000150:complications; D018451:Homosexuality, Male; D006801:Humans; D061346:Laser Capture Microdissection / Q000379:methods*; D008297:Male; D008875:Middle Aged; D027383:Papillomaviridae / Q000235:genetics*; D030361:Papillomavirus Infections / Q000150:complications*
chemical_list: D015415:Biomarkers
keywords: Anal cancer; Biomarkers; HSIL/HGAIN; Human papillomavirus; Laser capture microdissection; Precancerous conditions