Investigation of altered urinary metabolomic profiles of invasive ductal carcinoma of breast using targeted and untargeted approaches
Invasive ductal carcinoma (IDC) is a type of breast cancer, usually detected in advanced stages due to its asymptomatic nature which ultimately leads to low survival rate. Identification of urinary metabolic adaptations induced by IDC to understand the disease pathophysiology and monitor therapy response would be a helpful approach in clinical settings. Moreover, its non-invasive and cost effective strategy better suited to minimize apprehension among high risk population.
This study aims toward investigating the urinary metabolic alterations of IDC by targeted (LC-MRM/MS) and untargeted (GC–MS) approaches for the better understanding of the disease pathophysiology and monitoring therapy response.
Urinary metabolic alterations of IDC subjects (63) and control subjects (63) were explored by targeted (LC-MRM/MS) and untargeted (GC–MS) approaches. IDC specific urinary metabolomics signature was extracted by applying both univariate and multivariate statistical tools.
Statistical analysis identified 39 urinary metabolites with the highest contribution to metabolomic alterations specific to IDC. Out of which, 19 metabolites were identified from targeted LC-MRM/MS analysis, while 20 were identified from the untargeted GC–MS analysis. Receiver operator characteristic (ROC) curve analysis evidenced 6 most discriminatory metabolites from each type of approach that could differentiate between IDC subjects and controls with higher sensitivity and specificity. Furthermore, metabolic pathway analysis depicted several dysregulated pathways in IDC including sugar, amino acid, nucleotide metabolism, TCA cycle etc.
Overall, this study provides valuable inputs regarding altered urinary metabolites which improved our knowledge on urinary metabolomic alterations induced by IDC. Moreover, this study identified several dysregulated metabolic pathways which offer further insight into the disease pathophysiology.
KeywordsBreast cancer Invasive ductal carcinoma Urine Metabolomics LC-MRM/MS GC–MS
The authors are grateful to all the volunteers who participated in this study. THM acknowledges Department of Biotechnology, Govt. of India for fellowship. RT acknowledges Council of Scientific and Industrial Research, New Delhi, India for research associateship.
Conceived the study: THM, RT, SR; Designed the study: THM, RT, SR; Performed the experiments: THM, RT; Compiled the data: THM, RT, KT, VC, VN, SR; Analyzed the data and performed the multivariate statistical analysis and bioinformatics: THM, RT, KT, VC, VN, SR; Drafted the manuscript: THM, AM, SR; Provided clinical samples: AM; Provided chemicals and reagents: SR.
This research was supported by Department of Biotechnology, Govt. of India, India (RGYI Grant No. BT/PR6384/GBD/27/409/2012).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interests in relation to the work described.
The study was approved by the Ethics Committee of the Poona medical research foundation and National Centre for Cell Science (NCCS), Pune.
Fasting urine samples were collected with institutional review approval and after informed consent from all individual participants included in the study.
Research involving human participants and/or animals
All procedures performed were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.
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