Skip to main content

Advertisement

SpringerLink
Log in

Identification of serum metabolic markers for diagnosis of women with dormant genital tuberculosis

  • Original Article
  • Published:
Metabolomics Aims and scope Submit manuscript

Abstract

Introduction

Genital tuberculosis (GTB) in women is one of the common causes of infertility in emerging countries. As an intracellular pathogen, Mycobacterium tuberculosis in the endometrium significantly alters the host metabolism in dormant GTB cases. Nuclear magnetic resonance (NMR) based metabolic profiling has emerged as a useful tool for identification of biomarkers in biological fluids.

Objective

To investigate NMR based serum metabolic profile of dormant GTB women as compared to controls.

Methods

Dormant GTB women (n = 26) and unexplained infertile women (controls; n = 26), healthy proven fertile women undergoing voluntary sterilization (n = 25) and women undergoing recurrent spontaneous miscarriage (RSM) (n = 27) were included in the study. 700 MHz proton NMR spectra of serum collected from these patients were recorded. Multivariate analysis including principal component analysis, partial least squares discriminant analysis and orthogonal projection to latent structure-discriminant analysis was applied to all the spectra. Association of dysregulated serum metabolites with our earlier findings related to altered endometrial tissue metabolites in dormant GTB women was studied using multiple correlation analysis.

Results

This study indicates a clear metabolic differentiation between women with dormant GTB and controls. Metabolites including 3-hydroxybutyrate, succinate, citrate, acetate, l-glutamine, l-lysine, glutamate, l-threonine and 1-methyl histidine were found to be significantly upregulated in serum of women with dormant GTB compared with controls. Pearson’s correlation analysis showed a significant correlation between the expression of endometrial tissue and serum metabolites.

Conclusions

The set of identified metabolites may be considered as candidate markers for the diagnosis of dormant GTB and help clinicians in early therapeutic management.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Banerjee, P., Dutta, M., Srivastava, S., Joshi, M., Chakravarty, B., & Chaudhury, K. (2014). (1)H NMR serum metabonomics for understanding metabolic dysregulation in women with idiopathic recurrent spontaneous miscarriage during implantation window. Journal of Proteome Research, 13, 3100–3106.

    Article  CAS  PubMed  Google Scholar 

  • Banerjee, P., Jana, S. K., Pasricha, P., Ghosh, S., Chakravarty, B., & Chaudhury, K. (2013). Proinflammatory cytokines induced altered expression of cyclooxygenase-2 gene results in unreceptive endometrium in women with idiopathic recurrent spontaneous miscarriage. Fertility and Sterility, 99, 179–187.

    Article  CAS  PubMed  Google Scholar 

  • Bertini, I., Cacciatore, S., Jensen, B. V., Schou, J. V., Johansen, J. S., Kruhøffer, M., et al. (2012). Metabolomic NMR fingerprinting to identify and predict survival of patients with metastatic colorectal cancer. Cancer Research, 72, 356–364.

    Article  CAS  PubMed  Google Scholar 

  • Bhanu, N. V., Singh, U. B., Chakraborty, M., Suresh, N., Arora, J., Rana, T., et al. (2005). Improved diagnostic value of PCR in the diagnosis of female genital tuberculosis leading to infertility. Journal of Medical Microbiology, 54, 927–931.

    Article  CAS  PubMed  Google Scholar 

  • Bose, M. (2011). Female genital tract tuberculosis: how long will it elude diagnosis. Indian Journal of Medical Research, 134, 13–14.

    PubMed  PubMed Central  Google Scholar 

  • Bujak, R., Garcia-Alvarez, A., Ruperez, F. J., Nuno-Ayala, M., Garcia, A., Ruiz-Cabello, J., et al. (2014). Metabolomics reveals metabolite changes in acute pulmonary embolism. Journal of Proteome Research, 13, 805–816.

    Article  CAS  PubMed  Google Scholar 

  • Cao, M., Zhao, L., Chen, H., Xue, W., & Lin, D. (2012). NMR-based metabolomic analysis of human bladder cancer. Analytical Sciences, 28, 451–456.

    Article  CAS  PubMed  Google Scholar 

  • Dam, P., Shirazee, H. H., Goswami, S. K., Ghosh, S., Ganesh, A., Chaudhury, K., et al. (2006). Role of latent genital tuberculosis in repeated IVF failure in the Indian clinical setting. Gynecologic and Obstetric Investigation, 61, 223–227.

    Article  PubMed  Google Scholar 

  • Das, P., Ahuja, A., & Gupta, S. D. (2008). Incidence, etiopathogenesis and pathological aspects of genitourinary tuberculosis in India: A journey revisited. Indian Journal of Urology, 24, 356–361.

    Article  PubMed  PubMed Central  Google Scholar 

  • De Graaf, R. A., & Behar, K. L. (2003). Quantitative 1H NMR spectroscopy of blood plasma metabolites. Analytical Chemistry, 75, 2100–2104.

    Article  PubMed  Google Scholar 

  • Dutta, M., Joshi, M., Srivastava, S., Lodh, I., Chakravarty, B., & Chaudhury, K. (2012). A metabonomics approach as a means for identification of potential biomarkers for early diagnosis of endometriosis. Molecular BioSystems, 8, 3281–3287.

    Article  CAS  PubMed  Google Scholar 

  • Fernie, A. R., Trethewey, R. N., Krotzky, A. J., & Willmitzer, L. (2004). Metabolite profiling: From diagnostics to systems biology. Nature Reviews Molecular Cell Biology, 5, 763–769.

    Article  CAS  PubMed  Google Scholar 

  • Frediani, J. K., Jones, D. P., Tukvadze, N., Uppal, K., Sanikidze, E., Kipiani, M., et al. (2014). Plasma metabolomics in human pulmonary tuberculosis disease: A pilot study. PLoS ONE, 9, e108854.

    Article  PubMed  PubMed Central  Google Scholar 

  • Garris, D. R., & Smith, C. (1983). Diabetes-associated endometrial disruption in the ketonuric, diabetic Chinese hamster. Gynecologic and Obstetric Investigation, 16, 86–96.

    Article  CAS  PubMed  Google Scholar 

  • Hedjazi, L., Gauguier, D., Zalloua, P. A., Nicholson, J. K., Dumas, M. E., & Cazier, J. B. (2015). mQTL.NMR: an integrated suite for genetic mapping of quantitative variations of (1)H NMR-based metabolic profiles. Analytical Chemistry, 87, 4377–4384.

    Article  CAS  PubMed  Google Scholar 

  • Jindal, U. N. (2006). An algorithmic approach to female genital tuberculosis causing infertility. International Journal of Tuberculosis and Lung Disease, 10, 1045–1050.

    CAS  PubMed  Google Scholar 

  • Jindal, U. N., Bala, Y., Sodhi, S., Verma, S., & Jindal, S. (2010). Female genital tuberculosis: early diagnosis by laparoscopy and endometrial polymerase chain reaction. International Journal of Tuberculosis and Lung Disease, 14, 1629–1634.

    CAS  PubMed  Google Scholar 

  • Jindal, U. N., Verma, S., & Bala, Y. (2012). Favorable infertility outcomes following antitubercular treatment prescribed on the sole basis of a positive polymerase chain reaction test for endometrial tuberculosis. Human Reproduction, 27, 1368–1374.

    Article  CAS  PubMed  Google Scholar 

  • Karnovsky, A., Weymouth, T., Hull, T., Tarcea, V. G., Scardoni, G., Laudanna, C., et al. (2012). Metscape 2 bioinformatics tool for the analysis and visualization of metabolomics and gene expression data. Bioinformatics, 28, 373–380.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kostara, C. E., Papathanasiou, A., Psychogios, N., Cung, M. T., Elisaf, M. S., Goudevenos, J., et al. (2014). NMR-based lipidomic analysis of blood lipoproteins differentiates the progression of coronary heart disease. Journal of Proteome Research, 13, 2585–2598.

    Article  CAS  PubMed  Google Scholar 

  • Kulshrestha, V., Kriplani, A., Agarwal, N., Singh, U. B., & Rana, T. (2011). Genital tuberculosis among infertile women and fertility outcome after antitubercular therapy. International Journal of Gynecology and Obstetrics, 113, 229–234.

    Article  PubMed  Google Scholar 

  • Leroy, J. L., Vanholder, T., Opsomer, G., Van Soom, A., & de Kruif, A. (2006). The in vitro development of bovine oocytes after maturation in glucose and beta-hydroxybutyrate concentrations associated with negative energy balance in dairy cows. Reproduction in Domestic Animals, 41, 119–123.

    Article  CAS  PubMed  Google Scholar 

  • Lim, T. K., Gough, A., Chin, N. K., & Kumarasinghe, G. (2000). Relationship between estimated pretest probability and accuracy of automated Mycobacterium tuberculosis assay in smear-negative pulmonary tuberculosis. Chest, 118, 641–647.

    Article  CAS  PubMed  Google Scholar 

  • Liu, F., Yang, M., Wang, X., Yang, S., Gu, J., Zhou, J., et al. (2014). Acetylome analysis reveals diverse functions of lysine acetylation in Mycobacterium tuberculosis. Molecular and Cellular Proteomics, 13, 3352–3366.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Malik, S. (2003). Genital tuberculosis and implantation in assisted reproduction. Reviews in Gynaecological Practice, 3, 160–164.

    Article  Google Scholar 

  • Nambi, S., Gupta, K., Bhattacharyya, M., Ramakrishnan, P., Ravikumar, V., Siddiqui, N., et al. (2013). Cyclic AMP-dependent protein lysine acylation in mycobacteria regulates fatty acid and propionate metabolism. Journal of Biological Chemistry, 288, 14114–14124.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Paton, N. I., Chua, Y. K., Earnest, A., & Chee, C. B. (2004). Randomized controlled trial of nutritional supplementation in patients with newly diagnosed tuberculosis and wasting. American Journal of Clinical Nutrition, 80, 460–465.

    CAS  PubMed  Google Scholar 

  • Roy, H., & Roy, S. (2003). Use of polymerase chain reaction for diagnosis of endometrial tuberculosis in high risk subfertile women in an endemic zone. Journal of Obstetrics and Gynecology of India, 53, 260–263.

    Google Scholar 

  • Rozati, R., Roopa, S., & Naga Rajeshwari, C. (2006). Evaluation of women with infertility and genital tuberculosis. Journal of Obstetrics and Gynecology of India, 56, 423–426.

    CAS  Google Scholar 

  • Shin, J. H., Yang, J. Y., Jeon, B. Y., Yoon, Y. J., Cho, S. N., Kang, Y. H., et al. (2011). (1)H NMR-based metabolomic profiling in mice infected with mycobacterium tuberculosis. Journal of Proteome Research, 10, 2238–2247.

    Article  CAS  PubMed  Google Scholar 

  • Shrivastava, G., Bajpai, T., Bhatambare, G. S., & Patel, K. B. (2014). Genital tuberculosis: Comparative study of the diagnostic modalities. Journal of Human Reproductive Sciences, 7, 30–33.

    Google Scholar 

  • Somashekar, B. S., Amin, A. G., Rithner, C. D., Troudt, J., Basaraba, R., Izzo, A., et al. (2011). Metabolic profiling of lung granuloma in Mycobacterium tuberculosis infected guinea pigs: ex vivo 1H magic angle spinning NMR studies. Journal of Proteome Research, 10, 4186–4195.

    Article  CAS  PubMed  Google Scholar 

  • Somashekar, B. S., Amin, A. G., Tripathi, P., MacKinnon, N., Rithner, C. D., Shanley, C. A., et al. (2012). Metabolomic signatures in guinea pigs infected with epidemic-associated W-Beijing strains of Mycobacterium tuberculosis. Journal of Proteome Research, 11, 4873–4884.

    Article  CAS  PubMed  Google Scholar 

  • Subramani, E., Jothiramajayam, M., Dutta, M., Chakravorty, D., Joshi, M., Srivastava, S., et al. (2016a). NMR-based metabonomics for understanding the influence of dormant female genital tuberculosis on metabolism of the human endometrium. Human Reproduction, 31, 854–865.

    Article  CAS  PubMed  Google Scholar 

  • Subramani, E., Madogwe, E., Ray, C. D., Dutta, S. K., Chakravarty, B. N., Bordignon, V., et al. (2016b). Dysregulated LIF and its receptor regulated STAT3 pathway: A possible cause for repeated implantation failure in women with dormant genital tuberculosis? Fertility and Sterility, 105, 1076–1084.

    Article  CAS  PubMed  Google Scholar 

  • Thangappah, R. B. P., Paramasivan, C. N., & Narayanan, S. (2011). Evaluating PCR, culture and histopathology in the diagnosis of female genital tuberculosis. Indian Journal of Medical Research, 134, 40–46.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Weiner, J., Parida, S. K., Maertzdorf, J., Black, G. F., Repsilber, D., Telaar, A., et al. (2012). Biomarkers of inflammation, immunosuppression and stress with active disease are revealed by metabolomic profiling of tuberculosis patients. PLoS ONE, 7, e40221.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wevers, R. A., Engelke, U., & Heerschap, A. (1994). High-resolution 1H-NMR spectroscopy of blood plasma for metabolic studies. Clinical Chemistry, 40, 1245–1250.

    CAS  PubMed  Google Scholar 

  • Wishart, D. S., Knox, C., Guo, A. C., Eisner, R., Young, N., Gautam, B., et al. (2009). HMDB: A knowledgebase for the human metabolome. Nucleic Acids Research, 37, D603–D610.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhao, X., Chen, J., Ye, L., & Xum, G. (2014). Serum metabolomics study of the acute graft rejection in human renal transplantation based on liquid chromatography mass spectrometry. Journal of Proteome Research, 13, 2567–2659.

    Google Scholar 

  • Zhou, A., Ni, J., Xu, Z., Wang, Y., Lu, S., Sha, W., et al. (2013). Application of (1)H NMR spectroscopy-based metabolomics to sera of tuberculosis patients. Journal of Proteome Research, 12, 4642–4649.

    Article  CAS  PubMed  Google Scholar 

  • Zhu, Y., Guo, Z., Zhang, L., Zhang, Y., Chen, Y., Nan, J., et al. (2015). System-wide assembly of pathways and modules hierarchically reveal metabolic mechanism of cerebral ischemia. Scientific Reports, 5, 17068.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

The authors are thankful to all the volunteers participated in this study. One of the authors, E Subramani, is acknowledging to IIT Kharagpur and MHRD, Govt. of India for fellowship.

Funding

His work was partly funded by Indian Council of Medical Research, Govt. of India (No.5/7/372/09-RHN; KC).

Author information

Authors and Affiliations

  1. School of Medical Science and Technology, Indian Institute of Technology, Kharagpur, 721302, West Bengal, India

    Elavarasan Subramani, Mainak Dutta & Koel Chaudhury

  2. Cell Biology and Genetic Toxicology Laboratory, Centre of Advanced Study, Department of Botany, University of Calcutta, Kolkata, 700019, West Bengal, India

    Manivannan Jothiramajayam & Anita Mukherjee

  3. National Facility for High-field NMR, Tata Institute of Fundamental Research, Mumbai, 400005, Maharashtra, India

    Mamata Joshi & Sudha Srivastava

  4. Institute of Reproductive Medicine, Salt Lake, Kolkata, 700020, West Bengal, India

    Baidyanath Chakravarty

Authors
  1. Elavarasan Subramani
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mainak Dutta
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Manivannan Jothiramajayam
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mamata Joshi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sudha Srivastava
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Anita Mukherjee
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Baidyanath Chakravarty
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Koel Chaudhury
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Koel Chaudhury.

Ethics declarations

Conflicts of interest

Authors do not have any conflict of interest.

Ethical approval

All procedures performed in the present study involving human participants were in accordance with the ethical standards of the institutional and national research committee and with the 1964 Helsinki declaration and its later amendments.

Informed consent

Informed consent was obtained from all individual participants included in the study.

Electronic supplementary material

Below is the link to the electronic supplementary material.

11306_2016_1042_MOESM1_ESM.tif

Supplementary material 1 Representative T2-edited CPMG NMR spectra (δ 0.0−9.0) of serum from women with dormant GTB. Metabolites are indicated as follows: 1-TSP, 2-Lipids, 3-L-Isoleucine, 4-L-Leucine, 5-L-Valine, 6-3-Hydroxybutyric acid, 7-Lactate, 8-L-Alanine, 9-Adipic acid, 10-L-Lysine, 11-Acetate, 12-L-Proline, 13-n-Acetyl glycoproteins, 14-L-Glutamine, 15-Acetone, 16-D-Glutamate, 17-Citrate, 18-Creatine, 19-Creatinine, 20-Ornithine, 21-L-Carnitine, 22-D-Glucose, 23-Glycine, 24-L-Histidine, 25-L-Threonine, 26-L-Tyrosine, 27-L-Phenylalanine, 28-Formate, 29-Glycerol, 30-Betaine, 31-Succinate (TIFF 669 kb)

11306_2016_1042_MOESM2_ESM.tif

Supplementary material 2 Two-dimensional scatter plot of (A) PCA, (B) PLS-DA discriminates dormant GTB women, controls, proven fertile women and RSM cases. (C) Permutation test of PLS-DA model for validation (n = 200) (TIFF 1542 kb)

11306_2016_1042_MOESM3_ESM.tif

Supplementary material 3 Color map represents variable importance in projection (VIP) scores corresponding to the OPLS-DA model for dormant GTB and controls (TIFF 4830 kb)

11306_2016_1042_MOESM4_ESM.tif

Supplementary material 4 Scatter plot of OPLS-DA and its corresponding correlation coefficient plots obtained from (A) dormant GTB cases versus proven fertile women, (B) RSM cases versus proven fertile women (C) dormant GTB women versus RSM women (TIFF 2008 kb)

11306_2016_1042_MOESM5_ESM.tif

Supplementary material 5 The color bar represents VIP scores corresponding to OPLS-DA model of (A) dormant GTB cases versus proven fertile women, (B) RSM cases versus proven fertile women (C) dormant GTB women versus RSM women (TIFF 2711 kb)

11306_2016_1042_MOESM6_ESM.tif

Supplementary material 6 Compound network map of dysregulated metabolites. Blue hexagon represents input compound (potential metabolite markers). Pink hexagon indicated the related compounds and chemical reactions (TIFF 22000 kb)

Supplementary material 7 (DOCX 18 kb)

Supplementary material 8 (DOCX 28 kb)

Supplementary material 9 (DOCX 73 kb)

Supplementary material 10 (DOCX 18 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Subramani, E., Dutta, M., Jothiramajayam, M. et al. Identification of serum metabolic markers for diagnosis of women with dormant genital tuberculosis. Metabolomics 12, 99 (2016). https://doi.org/10.1007/s11306-016-1042-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s11306-016-1042-5

Keywords

Search

Navigation