Diagnostic panel of biomarkers for the assessment of sarcoidosis and tuberculosis identified using NMR based serum metabolomics approach
Creators: name: Kumar, Dinesh
name: Jain, Avinash
name: Kumar, Amit
name: Guleria, Anupam
name: Nath, Alok
name: Durga Prasanna Misra
name: Agarwal, Vikas
Publisher: name: F1000 Research Limited
Year: 2018

description: Tuberculosis (TB) and Sarcoidosis are two inflammatory granulomatous disorders and often pose a diagnostic challenge. Both mainly manifest in the lungs and diagnostic evaluation based on histopathological examination and culture, is often time consuming and results in delayed treatment. Sarcoidosis –a non-infectious granulomatous lung disease- may have diverse clinical impacts, ranging from benign to very severe; thus requires systemic treatment and subsequent monitoring as well [1,2]. Currently, there are no reliable clinical markers available to monitor management in such patients. Therefore, there is an emerging need to identify reliable clinical biomarkers for differential diagnosis of sarcoid from TB, their treatment monitoring and disease progression. The specific metabolic disturbances associated with sarcoid and TB will further help to understand the underlying disease mechanism so that the treatments can be tailored for rapid and efficient clinical management of these patients. Currently, the Department of Clinical Immunology, SGPGIMS Lucknow, in collaboration with Department of Pulmonary Medicines, SGPGIMS Lucknow and CBMR Scientists is making efforts to achieve both these objectives.<br><br>In recent past, several plasma/serum based metabolomics studies have been carried out involving sarcoid [3,4] or TB [5-11] cases, however, separately in different contexts; whereas comparative analysis between TB and sarcoid has never been performed to sought differential metabolic signatures. We hypothesized that metabolomics analysis of serum could reveal valuable biomarkers for differential diagnosis of these granulomatous lung diseases. Within this framework, we designed a study involving comparative analysis of serum metabolic profiles in patients with Sarcoid and TB. The study was approved by the institutional research and ethical committee and subsequently, the serum samples were obtained from Sarcoidosis (Mantoux negative Lofgren syndrome) patients attending the OPD of Department of clinical immunology, SGPGIMS, Lucknow and treatment naïve TB patients (proven by GenExpert, tissue diagnosis and/or by culture) attending the OPD of Department of Pulmonary Medicine, SGPGIMS, Lucknow, India. The enrolment of subjects was carried out according to the norms of World Medical Association (WMA) declaration of Helsinki. Overall, 35 sarcoidosis and 28 TB patients with mean age 40 ± 9.6 years and 30.3 ± 13.6 years and female: male ratio of 1.05:1 and 2.5:1, respectively were identified and enrolled in the study. To assess the disease specific metabolic disturbances, 54 normal healthy controls (NC, with mean age 34.2 ± 5.2 years and female: male ratios of 1:1) were also enrolled to validate the resulted metabolic perturbations through comparing with those reported previously. The serum metabolic profiles were measured using high resolution (800 MHz) 1D <sup>1</sup>H NMR (Nuclear Magnetic Resonance) spectroscopy and compared using advanced multivariate statistical analysis tools like partial least-squares discriminant analysis (PLS-DA) and orthogonal PLS-DA (OPLS-DA). The metabolites responsible for group separation were identified based on variable importance in projection (VIP) statistics. The resulted panel of discriminatory metabolites were further evaluated for statistical significance using independent student t-test and a p-value&lt;0.05 was used as a criterion for statistical significance. Consistent with various previous reports, the serum levels of glucose, glutamine, glyceryl phosphoryl choline (GPC) and lipid metabolites (LDL/VLDL, PUFA) were found to be decreased in TB patients compared to both Sarcoid and NC; whereas the serum levels of acetate, lactate, pyruvate, 3-hydroxybutyrate, isobutyrate, creatine, creatinine, citrulline and various amino acids (serine, threonine, arginine, lysine, leucine, isoleucine, lysine, glutamate, valine) were significantly increased. Overall, the results revealed that the TB patients exhibit significantly aberrant metabolic disturbances compared to both NC and Sarcoid patients thus can be screened with high sensitivity and specificity based on serum metabolomics analysis. These preliminary results will not only pave the way to develop an early and efficient diagnostic modality, but would further help to improve our understanding about infectious and non-infectious granulomatous lung diseases. 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