DOI: 10.1177/19450265261491098 ISSN: 1945-0265
Clinical Evaluation of miR-135b-5p and miR-21-5p Expression on Self-Collected Cervical Swab Samples for Triaging HR-HPV-Positive High-Grade Preneoplastic Lesions in India
Farhin Sultana, Ananya De, Sreeya Bose, Anirban Roychowdhury, Masmuda Khatun, Puja Chatterjee, Manisha Vernekar, Dipanwita Banerjee, Jayanta Chakrabarti, Ranajit Kumar Mandal, Chinmay Kumar Panda, Sankhadeep Dutta
Objective:
This study aimed to evaluate the triage performance of microRNA (miRNA) biomarkers on self-collected, high-risk human papillomavirus (HR-HPV)-positive cervical swabs to discern cervical intraepithelial neoplasia II–III or high-grade squamous intraepithelial lesions (HSIL) in Indian women for precise diagnosis and treatment prioritization.
Methods:
Through
in silico
analysis, miR-135b-5p and miR-21-5p were identified as upregulated and amplified miRNAs in cervical cancer (CaCx). Quantitative real-time PCR was used to assess miR-135b-5p and miR-21-5p and their targeting, LIM domain-containing protein 1 (LIMD1) and von Hippel–Lindau (VHL) messenger RNA (mRNA) expression, in a discovery cohort (
N
= 42; normal cervical epithelium [
N
= 9], CIN [
N
= 12], and CaCx [
N
= 21] tissue samples). Thereafter, miRNA expression was validated in an independent set of self-collected cervical swabs (
N
= 243; HR-HPV-positive [asymptomatic (
N
= 52), low-grade squamous intraepithelial lesions (LSIL) (
N
= 50), and HSIL (
N
= 52), CaCx (
N
= 35)] and HPV-negative normal [
N
= 54]). Receiver operating characteristic (ROC) curve analysis was performed to evaluate the diagnostic performance of these two candidate miRNAs.
Results:
In discovery-phase samples, miR-135b-5p showed progressive upregulation from CIN (
p
= 0.056) to CaCx (
p
= 0.012), while miR-21-5p showed significant overexpression at CIN (
p
= 0.011) and CaCx (
p
= 0.005) compared to normal tissue. Concurrently, significant downregulation of LIMD1/VHL mRNA expression was observed in the same CaCx samples. In the validation cohort, miR-135b-5p and miR-21-5p were significantly overexpressed in HSIL samples compared to lower-grade lesions (≤LSIL), with a corresponding downexpression of LIMD1/VHL. ROC curve analysis demonstrated high sensitivity and specificity for miR-21-5p and miR-135b-5p—both individually and in combination—for distinguishing HR-HPV + HSIL from LSIL, asymptomatic, or HPV-negative normal samples.
Conclusion:
miR-135b-5p and miR-21-5p constitute a feasible triage biomarker on noninvasive, self-collected cervical swab samples for early, precise diagnosis of women with HR-HPV-positive HSIL, requiring treatment prioritization in low-resource settings.