Estimating tissue-specific TNF mRNA levels prior to anti-TNFα treatment may support therapeutic optimisation in IBD patients

Abstract

Background and aims

Tumour necrosis factor-α (TNF) antagonists have improved the management of inflammatory bowel disease (IBD), however, their usage and administration persist to be suboptimal. Here, we examined the relationship between tissue-specific TNF mRNA expression in mucosal biopsies from IBD patients and anti-TNF treatment response.

Methods

Archived tissue samples from patients with luminal IBD that had all been or were in treatment with anti-TNF were included (18 adults and 24 paediatric patients). Patients were stratified into three groups according to anti-TNF response: responders, primary non-responders (PNR) and secondary loss of response (SLOR). TNF mRNA was detected using RNAscope in situ hybridisation (ISH) and the expression was quantified using image analysis.

Results

The ISH analysis showed varying occurrence of TNF mRNA positive cells located in lamina propria and often with increased density in lymphoid follicles (LF). Consequently, expression estimates were obtained in whole tissue areas with and without LF. Significantly higher TNF mRNA expression levels were measured in adults compared to paediatric patients in both the analyses with and without LF (p = .015 and p = .016, respectively). Considering the relation to response, the adult and paediatric patients were evaluated separately. In adults, the TNF expression estimates were higher in PNRs compared to responders with and without LF (p = .017 and p = .024, respectively).

Conclusion

Our data indicate that adult PNR have significantly higher TNF mRNA levels than responders. This suggests that higher anti-TNF dose may be considered for IBD patients with high TNF mRNA expression estimates from the start of treatment.

Graphical Abstract

Keywords:

• Image analysis
• lymphoid follicle
• mucosal TNF mRNA
• Paediatric IBD
• quantitative in situ hybridisation
• inflammation
• treatment non-response
• Tumour necrosis factor

Acknowledgements

We would like to thank laboratory technicians Trine Møller, Bioneer A/S for her valuable technical assistance.

Ethics statement

The study was conducted in accordance with the ethical standards, according to the Declaration of Helsinki and according to national and international guidelines. The study was approved by the Danish National Committee on Health Research Ethics (H-20032221) and the Danish Data Protection Agency (P-2020-737).

Authors contributions

No additional writing assistance was used for this manuscript. L.B.R., B.S.N., J.P.J., E.L., M.M. and E.H. all contributed to the conceptualisation and the design of study. L.B.R., E.L., M.M. and J.P.J collected samples, B.S.N., L.B.R. and J.P.J. designed experiments and analysed the data. J.P.J., L.B.R. and B.S.N. drafted the manuscript. E.L., M.M. and E.H. critically revised the manuscript for important intellectual content. All authors have approved the final version of the manuscript.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data availability statement

The data underlying this article are available in the article and in its online supplementary material.

Additional information

Funding

This work was supported by Aage Louis Hansen fonden [J.nr.20-2B-5995]; Herlev internal research funding [after year, 2019]; Colitis-Crohn Foreningen [2021]; Torben og Alice Frimodts Fond [FK1900]; Aase and Ejnar Danielsen’s Foundation [20-10-0411]. The opinions stated in this manuscript are those of the authors own and do not necessarily reflect those of the organisation or funder.