Cytological Immunostaining of HMGA2, LRP1B, and TP63 as Potential Biomarkers for Triaging Human Papillomavirus-Positive Women

Abstract

Background: Since human papillomavirus (HPV) DNA testing has been promoted as primary screening strategy, the triage method has also evolved from morphological testing to a molecular biomarker detection to improve screening efficiency. In this study, we investigated the performance of three HPV integration hot-spots, HMGA2, LRP1B, and TP63, as potential triage markers in HPV screening tests. Materials and Methods: This cross-sectional study was conducted from November 2016 to December 2017 in the First Affiliated Hospital of Sun Yat-sen University. Immunocytochemistry was carried out using residual cervical cell samples from 121 HPV-positive cases (23 normal, 24 cervical intraepithelial neoplasia (CIN) 1, and 74 CIN2+). Results: Of the 121 cases, 77 showed completely paired for the three biomarkers. In these 77 cases, receiver operating characteristic (ROC) analysis of HMGA2 showed the best potential for detecting CIN2+ among HPV+ cases (sensitivity 70%; specificity 91.89%; AUC 0.839). TP63 was second most effective biomarker (AUC 0.838; sensitivity 80%; specificity 81.08%). In contrast, LRP1B had the smallest AUC (0.801) among the three biomarkers but had the highest sensitivity (90%) and specificity (56.76%). To test the triage value of combining the three biomarkers, logistic regression was conducted followed by ROC comparison analysis. Promisingly, the combination of the three biomarkers gave the largest AUC of 0.951 with 92.5% sensitivity and 89.1% specificity ( P < .0001 compared to liquid-based cytology test by Z -test). Conclusions: A combination of HMGA2, LRP1B, and TP63 as potential biomarkers may be useful for screening during triage of HPV-positive patients, particularly for detecting CIN2 + .

LRP1B mutation is associated with tumor HPV status and promotes poor disease outcomes with a higher mutation count in HPV-related cervical carcinoma and head & neck squamous cell carcinoma

“…LRP1B was also detected as an HPV integration hotspot in CC [27], as suggested by our results, HPV-LRP1B integration was associated with the downregulation of LRP1B. Additionally, the LRP1B IHC staining score could serve as a sensitive biomarker for cervical lesion detection, with 0.801 area under curve (AUC), 90% sensitivity, and 56.76% specificity [39]. In our results, the expression status of LRP1B was identified as a biomarker for normal and neoplasm samples of CC and HNSCC, with AUC 0.8007 in HNSCC and 0.9024 in CC, which suggested that LRP1B could serve as a biomarker in CC and HNSCC.…”

“…(B) E6 (total/spliced/un-spliced) expression in HPV 16 integration samples and HPV non-integration samples. (C) Distribution of HPV-LRP1B integration samples in a cervical carcinoma cohort [39]. (D) IHC staining score of LRP1B in cervical carcinoma samples with different integration sites or normal samples from the same cohort [39].…”

Accurate Detection of HPV Integration Sites in Cervical Cancer Samples Using the Nanopore MinION Sequencer Without Error Correction

“…The significantly increased gene expression of HMGA2 and TP63 has been reported in neoplastic samples where HPV is integrated into their introns, whereas LRP1B is under expression with HPV integration in their flanking region ( Michael et al, 2014 ; Hu et al, 2015 ). Therefore, a combination of HMGA2, LRP1B , and TP63 as potential biomarkers may be useful for screening during triage of HPV-positive patients, particularly for detecting CIN2+ ( Jiang et al, 2019 ).…”

“… Ibragimova et al (2018) and Koneva et al (2018) discovered that patients with integrated HPV status have a worse survival rate compared to those with episomal HPV. The increasing number of studies about HPV integration has stimulated the idea that HPV integration status may be a biomarker for the diagnosis, progression, and survival prediction, and even as a biomarker for cancer screening ( Han et al, 2015 ; Liu et al, 2018 ; Grayson et al, 2019 ; Jiang et al, 2019 ). Harlé et al (2019) found the same integration sites and DNA deletion regions between tongue and anal cancer of the same patient.…”

Discovery of key genes as novel biomarkers specifically associated with HPV-negative cervical cancer

“…The architectural transcription factor HMGA2 is a nonhistone DNAbinding factor that tightly binds to adenine and thymine (AT)-rich sequences in the minor groove of the DNA helix. 19 Jiang et al 20 identified that HMGA2 combined with LRP1B and TP63 function as potential biomarkers that are useful for triaging HPV-positive patients, especially CIN2+ patients. Notably, the depletion of HMGA2 counteracted the epithelial-mesenchymal transition and lymph node metastasis by suppressing the attenuated total reflectance (ATR)/Chk1 signaling pathway in cervical cancer.…”