Abstract
Background
Gestational diabetes mellitus (GDM) is a common pregnancy complication associated with hyperglycaemia, chronic inflammation and adverse health outcomes. Regulatory T cells (Tregs) are thought to contribute to GDM due to their role in suppressing inflammation. However, whether specific Treg subsets are transcriptionally dysregulated in patients with GDM remains unclear.
Methods
To investigate Treg transcriptional variation in GDM, we applied single-cell RNA sequencing to Tregs and CD4 + T cells isolated from the blood of 13 healthy pregnant women and 10 female patients with GDM.
Results
We observed no significant differences in Treg cluster proportions with disease status, however, Memory CD4 + T cells were more abundant in patients diagnosed with GDM, substantiated by mass cytometry. We report Treg subsets altered in GDM, including naive Tregs with reduced expression of AP-1 transcription factor subunits and effector Tregs with increased signalling of genes associated with angiogenesis. Expression levels of genes dysregulated in GDM Tregs were informative of GDM status in pseudobulk, placental and whole blood mRNA from independent cohorts. TXNIP, which regulates glucose levels, emerged as the most significant discriminator of GDM status from bulk mRNA.
Conclusions
This study uncovers transcriptional differences of Treg cell subsets from GDM patients and transcriptional markers informative of GDM status.
Plain Language Summary
Gestational diabetes mellitus (GDM) is a common pregnancy condition linked to high blood sugar and increased inflammation, which can affect the health of both mother and baby. Immune cells called regulatory T cells (Tregs) help control inflammation, and their activity in a mother’s blood may be linked to GDM. To understand how Tregs behave in patients with GDM, we captured these cells from blood samples of pregnant women diagnosed with GDM and pregnant women without a GDM diagnosis. We profiled the expression of RNA in individual Tregs from these patients. We found that, while overall Treg numbers are similar, the activity of specific genes varies in Tregs from women with GDM. Disrupted RNA levels of one gene related to glucose control (TXNIP) may be an informative marker for GDM in blood. Our findings enhance the understanding of immune changes in GDM and may inform future approaches for early detection and monitoring.
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Data availability
Single-cell RNA sequencing data has been submitted to Gene Expression Omnibus (Accession: GSE280975). Bulk RNA sequencing data was downloaded from Gene Expression Omnibus (Accession: GSE154414; GSE92772 - RNA sequencing data of whole blood cells of normal glucose tolerant (NGT) and gestational diabetes (GDM) pregnant women). Data underlying the figures are available on Zenodo59.
Code availability
R scripts used to perform the analyses are available at GitHub60.
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Acknowledgements
This research is supported by the Foetal Medicine Foundation (registered charity 1037116 Project Number 909375), and the Carlsberg Foundation (CF23-0418). PS is supported by an NIHR Clinical Lectureship (CL-2018-17-002) an Academy of Medical Sciences Starter Grant for Clinical Lecturers (SGL023\1023). The project was also supported by the National Institute for Health Research (NIHR) Biomedical Research Centre at Guy’s and St Thomas’ National Health Service Foundation Trust and King’s College London (IS-BRC-1215–20006). We would like to thank the team and patients at King’s College Hospital who donated their blood for this project. We sincerely thank the team at the Advanced Cytometry Platform, R&D Department and the Genomics facility at Guy’s and St Thomas’ NHS Foundation Trust, Guy’s Hospital, London.
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N.E.M.*, N.M.*, A.E.* and M.T.M.M* contributed equally to this work. N.E.M.*: Data analysis, manuscript writing, manuscript editing. A.E.*: Consent, sample collection, and data analysis. N.M.: Consent, sample collection, and data analysis. M.T.M.M.*: Data analysis, manuscript editing. S.K.: Data analysis. H.V.: Conducted experiments. A.B.: Data analysis and manuscript editing. A.M.: Data analysis. T.T.: Provided expert opinion. G.L.: Provided expert opinion and edited the manuscript. P.D.: Data analysis and provided expert opinion. K.H.N.: Provided expert opinion and edited the manuscript. C.S.: Provided expert opinion and edited the manuscript. P.S.: Conceptualized the study, conducted experiments, collected samples, wrote and edited the manuscript.
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Communications Medicine thanks Jacob Friedman, Peijie Zhou and Joan Camuñas-Soler for their contribution to the peer review of this work. A peer review file is available.
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Mensah, N.E., Efthymiou, A., Mureanu, N. et al. Single-cell transcriptomics identifies regulatory T cell heterogeneity in gestational diabetes mellitus. Commun Med (2026). https://doi.org/10.1038/s43856-026-01563-0
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DOI: https://doi.org/10.1038/s43856-026-01563-0
