title: Evaluation of HPV infection and smoking status impacts on cell proliferation in epithelial layers of cervical neoplasia.
abstract: Accurate cervical intra-epithelial neoplasia (CIN) lesion grading is needed for effective patient management. We applied computer-assisted scanning and analytic approaches to immuno-stained CIN lesion sections to more accurately delineate disease states and decipher cell proliferation impacts from HPV and smoking within individual epithelial layers. A patient cohort undergoing cervical screening was identified (n = 196) and biopsies of varying disease grades and with intact basement membranes and epithelial layers were obtained (n = 261). Specimens were sectioned, stained (Mib1), and scanned using a high-resolution imaging system. We achieved semi-automated delineation of proliferation status and epithelial cell layers using Otsu segmentation, manual image review, Voronoi tessellation, and immuno-staining. Data were interrogated against known status for HPV infection, smoking, and disease grade. We observed increased cell proliferation and decreased epithelial thickness with increased disease grade (when analyzing the epithelium at full thickness). Analysis within individual cell layers showed a ≥50% increase in cell proliferation for CIN2 vs. CIN1 lesions in higher epithelial layers (with minimal differences seen in basal/parabasal layers). Higher rates of proliferation for HPV-positive vs. -negative cases were seen in epithelial layers beyond the basal/parabasal layers in normal and CIN1 tissues. Comparing smokers vs. non-smokers, we observed increased cell proliferation in parabasal (low and high grade lesions) and basal layers (high grade only). In sum, we report CIN grade-specific differences in cell proliferation within individual epithelial layers. We also show HPV and smoking impacts on cell layer-specific proliferation. Our findings yield insight into CIN progression biology and demonstrate that rigorous, semi-automated imaging of histopathological specimens may be applied to improve disease grading accuracy.
authors.affiliation: Department of Integrative Oncology, British Columbia Cancer Research Centre, Vancouver, British Columbia, Canada.
authors.forename: Martial
authors.initials: M
authors.lastname: Guillaud
authors.affiliation: Department of Integrative Oncology, British Columbia Cancer Research Centre, Vancouver, British Columbia, Canada.
authors.forename: Timon P H
authors.initials: TP
authors.lastname: Buys
authors.affiliation: Department of Integrative Oncology, British Columbia Cancer Research Centre, Vancouver, British Columbia, Canada.
authors.forename: Anita
authors.initials: A
authors.lastname: Carraro
authors.affiliation: Department of Integrative Oncology, British Columbia Cancer Research Centre, Vancouver, British Columbia, Canada.
authors.forename: Jagoda
authors.initials: J
authors.lastname: Korbelik
authors.affiliation: Department of Obstetrics and Gynecology, Paul L. Foster School of Medicine, Texas Tech University Health Sciences Center at El Paso, El Paso, Texas, United States of America.
authors.forename: Michele
authors.initials: M
authors.lastname: Follen
authors.affiliation: Department of Pediatrics, Dan L. Duncan Cancer Center, Baylor College of Medicine, Houston, Texas, United States of America.
authors.forename: Michael
authors.initials: M
authors.lastname: Scheurer
authors.affiliation: Department of Diagnostic Sciences, School of Dentistry, University of Texas Health Science Center, Houston, Texas, United States of America.
authors.forename: Karen Adler
authors.initials: KA
authors.lastname: Storthz
authors.affiliation: Department of Pathology, British Columbia Cancer Agency, Vancouver, British Columbia, Canada.
authors.forename: Dirk
authors.initials: D
authors.lastname: van Niekerk
authors.affiliation: Department of Integrative Oncology, British Columbia Cancer Research Centre, Vancouver, British Columbia, Canada.
authors.forename: Calum E
authors.initials: CE
authors.lastname: MacAulay
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D001485:Basement Membrane / Q000473:pathology*; D014408:Biomarkers, Tumor; D001706:Biopsy; D049109:Cell Proliferation; D004848:Epithelium / Q000473:pathology; D005260:Female; D006801:Humans; D019394:Ki-67 Antigen / Q000096:biosynthesis; D008875:Middle Aged; D060787:Neoplasm Grading; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D012907:Smoking / Q000473:pathology; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis* / Q000473:pathology / Q000821:virology
chemical_list: D014408:Biomarkers, Tumor; D019394:Ki-67 Antigen