368P - Distinct gene expression profiling explored using nanostring tumor signalling 360 panel with validations in different clinical stages of oral submucous fibrosis patients: A first Indian study

Background

Oral Submucous Fibrosis (OSMF) is a chronic debilitating disease more frequently found in the South East Asian population. This disease poses a public health priority, as it is grouped under oral potentially malignant disorders, with malignant transformation rates of around 7 to 13%. Hence, early identification of high-risk OSMF patients is of the utmost importance to prevent malignant transformation. mRNA expression profiling is a promising method for identifying differentially expressed genes for disease prognosis in OSMF. The genetic profiling was performed using Tumor Signaling (TS) 360 Panel (Nanostring platform) to profile 780 humans across 40+ annotated pathways.

Methods

The RNA was initially isolated from patient tissue samples in different clinical stages of OSMF (n=8), OSMF transformed into oral squamous cell carcinoma (OSCC) (n=5) and healthy controls (HC) (n=5). The analysis of gene expression was conducted on the nCounter® TS 360™ Panel and NanoString platform. The raw transcriptome data were subjected to housekeeping-gene normalization using the geNorm algorithm in nCounter Advanced Analysis ver. 2.0.115. Normalized data were log2-transformed for analysis. A quality check of raw data was conducted using nSolver Analysis Software ver. 4.0 and NanoStringQCpro ver. 1.14.0.

Results

Among the 780 genes, AR, RPTOR and PRDX6 showed the highest differential expression between OSMF and OSCC (2.48, 1.1 and 0.76 fold change, respectively; p < 0.05). While, MLANA, WEE1 and MYB showed the highest differential expression between HC and OSCC (5.16, 2.61 and 2.11 fold change, respectively; p < 0.05). The upregulated genes were further validated using real time PCR which showed significant upregulation in OSMF and OSCC.

Conclusions

The present study is the first of its kind in India to the best of our knowledge, assessing the gene expression using the Nanostring platform in different clinical stages of OSMF. with validation in a large series of cases. The currenty study has evolved a panel of biomarkers, namely PRDX6, MLANA and AR to be potentially useful in identifying high-risk OSMF patients with an increased risk of OSCC development.

Clinical trial identification

Editorial acknowledgement

Legal entity responsible for the study

C.V. Divyambika; R. Vijayalakshmi.

Funding

Indian Council of Medical Research (ICMR): 5/4/2-4/Oral Health/2021/NCD-II.

Disclosure

All authors have declared no conflicts of interest.