Discovery of novel metabolic signatures for early identification of women at risk of developing gestational hypertension.

INTRODUCTION: Gestational hypertension (GH) is defined as the presence of systolic blood pressure (BP) ≥ 140 mm Hg and/or diastolic BP ≥ 90 mm Hg, measured at least 4 h apart after 20 weeks of gestation. Early identification of women at high-risk of developing GH could contribute significantly towards improved maternal and fetal outcomes. OBJECTIVES: To determine early metabolic biomarkers in women with GH as compared with normotensive women. METHODS: Serum samples were collected from subjects during three stages of their pregnancy: 8-12 weeks, 18-20 weeks and after 28 weeks (< 36 weeks) of gestation and studied using nuclear magnetic resonance (NMR) metabolomics approach. Multivariate and univariate analyses were performed to determine the significantly altered metabolites in GH women. RESULTS: A total of 10 metabolites, including isoleucine, glutamine, lysine, proline, histidine, phenylalanine, alanine, carnitine, N-acetyl glycoprotein and lactic acid were observed to be significantly downregulated during all pregnancy stages in women with GH as compared with controls. Furthermore, expression of 5 metabolites in the first trimester i.e., phenylalanine [area under the curve (AUC) = 0.745], histidine [AUC = 0.729], proline [AUC = 0.722], lactic acid [AUC = 0.722], and carnitine [AUC = 0.714] exhibited highest potential in discriminating GH from normotensive women. CONCLUSION: The present study is the first of its kind to identify significantly altered metabolites that have the potential to discriminate between women at risk of developing GH and normotensive women across three trimesters of pregnancy. This opens up the possibility of exploring these metabolites as potential early predictive markers of GH.

Author Correction: Metabolomics reveals perturbations in endometrium and serum of minimal and mild endometriosis.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Metabolomics reveals perturbations in endometrium and serum of minimal and mild endometriosis.

Endometriosis is a common benign gynecological disease, characterized by growth and proliferation of endometrial glands and stroma outside the uterus. With studies showing metabolic changes in various biofluids of endometriosis women, we have set upon to investigate whether endometrial tissue show differences in their metabolic profiles. 1H NMR analysis was performed on eutopic endometrial tissue of women with endometriosis and controls. Analysis was performed on spectral data and on relative concentrations of metabolites obtained from spectra using multivariate and univariate data analysis. Analysis shows that various energy, ketogenic and glucogenic metabolites have significant altered concentrations in various stages of endometriosis. In addition, altered tissue metabolites in minimal and mild stages of endometriosis were explored in serum of these patients to assess their role in disease diagnosis. For Stage I diagnosis alanine was found to have 90% sensitivity (true positives) and 58% specificity (true negatives). For Stage II diagnosis alanine, leucine, lysine, proline and phenylalanine showed significant altered levels in serum. While sensitivity of these serum metabolites varied between 69.2-100% the specificity values ranged between 58.3-91.7%. Further, a regression model generated with this panel of serum markers showed an improved sensitivity and specificity of 100% and 83%, respectively for Stage II diagnosis.

Mycobacterial heat shock protein 65 mediated metabolic shift in decidualization of human endometrial stromal cells.

Successful implantation is dependent on the appropriate decidualization of endometrial stromal cells for the establishment of pregnancy in women. Mycobacterial heat shock protein 65 (HSP65) is involved in pathogenesis of the genital tuberculosis (GTB), one of the common causes of infertility in emerging countries. Though implantation failure appears to be the major cause, understanding the status of decidualizaiton process in women diagnosed with GTB has not been thoroughly addressed. We, therefore, explored the effect of HSP65 protein on the endometrial cell metabolism during in vitro decidualization. In order to identify the cellular metabolism of decidual cells with and without HSP65 treatment, proton NMR based characterization of metabolites extracted from cells and culture media were performed. In presence of HSP65, significant reduction in the decidual phenotype of endometrial stromal cells and prolactin expression is suggestive of impairment in decidualization. The intracellular and extracellular metabolic changes in HSP65 treated endometrial stromal cells produced a distinct pattern, reflecting the interaction between the protein and cellular metabolism. HSP65 mediated dysregulation in cellular metabolism is associated with poor decidualization. Besides enriching the present knowledge on metabolic changes underlying stromal cells decidualization, these findings assist in identifying potential molecular causes for decidualization failure in GTB women.

Dysregulated leukemia inhibitory factor and its receptor regulated signal transducers and activators of transcription 3 pathway: a possible cause for repeated implantation failure in women with dormant genital tuberculosis?

OBJECTIVE: To investigate the influence of dormant Mycobacterium tuberculosis on the expression of various endometrial receptivity markers and leukemia inhibitory factor (LIF)-signal transducers and activators of transcription 3 (STAT3) signaling pathway. Expression of endometrial receptivity markers and LIF-STAT3 signaling in in vitro decidualized human endometrial stromal cells (hESC) treated with 65 kDa mycobacterial heat shock protein (HSP65) is also explored. DESIGN: A prospective study. SETTING: Tertiary care hospital and reproductive health research unit. PATIENT(S): Endometrial tissue samples were collected from 38 women who tested positive for Mycobacterium tuberculosis and 30 normal women with proven fertility undergoing sterilization. In vitro decidualization of hESC was performed. INTERVENTION(S): Endometrial biopsies collected from all women during implantation window and treatment of hESC with HSP65. MAIN OUTCOME MEASURE(S): Measurement of various endometrial receptivity markers including αvß3 integrin, E-cadherin, MECA-79, mucin-1, and pinopodes and LIF/LIFR-STAT3 signaling molecules expressed in the endometrium of women with dormant genital tuberculosis (GTB) during implantation window and measured also in HSP65-treated hESC. RESULT(S): Significantly reduced levels of endometrial receptivity markers LIF, LIFR, and pSTAT3 were observed in endometrium of women with dormant GTB as compared with controls. A similar trend was observed under in vitro conditions with decreased level of phosphorylated STAT3 in HSP65-treated hESC. However, no change in the expression of endometrial receptivity markers under in vitro conditions was observed. CONCLUSION(S): Our findings suggest that endometrium of women with dormant GTB is associated with poor receptivity, as evidenced by reduced receptivity markers and aberrant LIF-STAT3 signaling. In vitro treatment of hESC with HSP65 also confirms compromised endometrial decidualization.

Mass spectrometry and bioinformatics analysis data.

2DE and 2D-DIGE based proteomics analysis of serum from women with endometriosis revealed several proteins to be dysregulated. A complete list of these proteins along with their mass spectrometry data and subsequent bioinformatics analysis are presented here. The data is related to "Investigation of serum proteome alterations in human endometriosis" by Dutta et al. [1].

Investigation of serum proteome alterations in human endometriosis.

Endometriosis is a common benign gynecological disease, characterized by proliferation of functional endometrial glands and stroma outside the uterine cavity. The present study involves investigation of alterations in the serum proteome of endometriosis patients compared to healthy controls using 2DE and 2D-DIGE combined with MALDI TOF/TOF-MS. Comparison of serum proteome of endometriosis patients and healthy subjects revealed 25 significant differentially expressed proteins. Gene ontology and network analysis, performed using PANTHER, DAVID, WebGestalt and STRING, revealed that the differentially expressed proteins are majorly involved in response to stimulus, immune system, metabolic, localization and cellular processes. For serum diagnostic marker identification, several robust statistical screening procedures were applied to identify the set of the most significant proteins responsible for successful diagnosis of different endometriosis stages. Partial least squares (PLS) based marker selection tool and orthogonal partial least squares-discriminant analysis (OPLS-DA) were used to identify the most significant proteins for disease prediction. Western blotting validation in a separate cohort of patients revealed that haptoglobin (HP), Ig kappa chain C region (IGKC), alpha-1B-glycoprotein (A1BG) can be considered effective serum protein markers for the diagnosis of Stage II, III and IV endometriosis. For diagnosis of Stage I, only IGKC and HP seemed promising. BIOLOGICAL SIGNIFICANCE: Globally, about 12 in 100 women of reproductive age are diagnosed with endometriosis. The pathogenesis of the disease still remains unclear, leading to non-specific therapeutic approaches for disease management. Moreover, there is a delay of 8-12years in correct diagnosis after the initial onset of symptoms leading to a considerable impact on the woman's lifestyle. Also, the gold standard for diagnosis of endometriosis, laparoscopy, is an invasive procedure. The value of a noninvasive or semi-invasive diagnostic test for endometriosis with easily accessible fluids such as plasma, serum, urine, and saliva is, therefore, rightfully recognized. The present study is expected to considerably improve the understanding of the disease pathogenesis along with improved diagnostics and therapeutic approaches leading to better management of the disease.

Altered metabolic profile in early and late onset preeclampsia: An FTIR spectroscopic study.

OBJECTIVE: Metabolic anomalies, if any, between early and late onset preeclampsia [PE] were explored using Fourier transform infrared [FTIR] spectroscopy. SETTING: Department of Gynecology and Obstetrics, SSKM Hospital, IPGMER, Kolkata and Midnapur Medical College Hospital, Midnapur, India. SAMPLE: 80 pregnant women attending routine antenatal care units; (i) early onset PE [gestational age; GA<34weeks] (ii) late onset PE [GA>34weeks] (iii) early onset control [GA 24-34weeks] and (iv) late onset control [GA>34weeks]. METHODS: Serum FTIR spectra were obtained in the wave-number range of 600-4000cm(-1) at 4cm(-1) resolution. (1)H NMR and estimation of atherosclerotic index (AI) were performed to validate the FTIR findings. MAIN OUTCOME MEASURE(S): Clinical characteristics and metabolic profile. RESULTS: 13 spectral peaks corresponding to the carbohydrate, protein and lipid region were significantly altered in early onset PE [P<0.001; at 95% confidence interval]. Discriminant analysis identified five highly significant wave-numbers (1078, 1088, 1122, 1169 and 1171cm(-1)) having ⩾80% overall accuracy. Hierarchical cluster analysis of the obtained spectra at these 5 wave-numbers provided excellent segregation of early and late onset PE with respect to their controls. Principal component analysis revealed that these 5 wave-numbers significantly separated the two sub-groups of PE (97.95% of the total variance). (1)H NMR results showed that serum levels of glutamate, choline, alanine and lactate were significantly higher while ariginine and citrate were significantly decreased in early onset PE as compared to late onset cases. CONCLUSION: Our study reveals differences in metabolomic profiles of early and late onset preeclamptic cases.