Spotlight: Use of RNA-Seq transcriptome profiling to identify the immune response and inflammation markers in latent endometrial tuberculosis

Introduction

Dai et al (2025) deployed a multi-faceted approach to investigate the mechanisms controlling Latent Endometrial Tuberculosis (LETB).

LETB damages endometrial structure and function, undermining implantation and increasing infertility risk.

LETB can be asymptomatic and remain undetected until there are problems such as pregnancy loss and so early detection is essential.

Current diagnostics lack specificity, creating a need for molecular markers that distinguish LETB from active TB and healthy endometrium. There is an urgent need for new, more reliable diagnostic approaches.

Main Points

• Traditional diagnostic methods often fail to accurately identify latent infections, thereby complicating the detection of LETB.
• TB latency and progression are driven by host immune responses.
• The current study investigated women aged 20 to 40 who underwent IVF treatment due to tubal factors. TB Interferon Gamma Release Assays (TB-IGRA) testing was conducted on endometrial biopsies, along with CD38 and CD138 labelling to assess the level of inflammation.
• There was no difference in TB-IGRA results between control and LETB patients – inflammatory (IG) and non-inflammatory groups (NIG). There were also no differences noted between groups when ultrasound images, Haematoxylin and Eosin (H&E) staining or immunohistochemistry for CD38 & CD138.
• Analysis of RNA-Seq data revealed common pool of 80 differentially expressed genes in the control, LETB, and TB groups. Shared inflammatory genes in the CG-NIG and CG-IG data reduced this to 79, from which 21 genes were identified as LETB-specific inflammatory genes.
• Seven candidate genes (IFI30, HCK, SPI1, IL1B, ITGB2, FCGR2A) were analysed using bioinformatic approaches including Gene Ontology (GO) which revealed that the genes of interest were involved in key immune responses. Using the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis this geneset were further investigated to indicate the pathways that were key immune cell signal transduction drivers for inflammatory and immune responses in LETB.
• Further validation of these genes by qPCR revealed significantly higher expression in the IG group. H&E staining validation showed structural alterations in the IG, and IHC validation using HCK and ITGB2 exhibited significant enhancement to the IG group, all in comparison to the NIG group

Conclusion

RNA-Seq was used to identify genetic mechanisms driving LETB disease progression, with the genes playing key roles in immune pathways. The data was further verified using various proteomic and statistical techniques, to confirm the genes as having the potential to be deployed as diagnostic biomarkers for LETB, decreasing the risks associated with IVF, and inflammatory disorders such as arthritis.

EPISTEM SERVICES

Epistem regularly deploys a wide spectrum of techniques to interrogate the transcriptome, proteome and informatic output:

• Transcriptomic Analysis for gene/pathway discovery via RNA-Seq (RNA-Seq) and confirmation by qPCR. NGS link
• Powerful bioinformatic analysis to create robust gene-signatures for biomarker discovery. Bioinformatics link
• Spatial location of active transcription by RNAscope or Laser Capture Microscopy.
• Inflammatory cell profiling via flow cytometry and cytokine profiling via multiplex analysis and ELISA based assays

• Protein localisation via immunohistochemistry. IHC link