Introduction
The highest risk factor of inflammatory bowel disease (IBD) is having a first-degree relative (FDR) with IBD. However, genetic variation accounts for only a small proportion of IBD heritability. Evidence from family and twin studies underscores the role of shared environment for the development of IBD. In a recent study based on two population-based IBD cohorts from South-Eastern Norway, thirty years apart, we reported a substantial increase in the rate of familial IBD (1).
Aims & Methods
This study investigates the potential underlying mechanisms for the increased rate of familial IBD in South-Eastern Norway.
We examine the association between shared environment and gene expression across IBD phenotypes [Crohn’s disease (CD) and ulcerative colitis (UC)] and four comparison groups: (I) multiplex IBD familial patients (≥3 FDRs with IBD), (II) healthy relatives in multiplex IBD families, (III) sporadic IBD patients, and (IV) symptomatic non-IBD controls. Expression analyses focus on four candidate genes, FKBP5, BCL3, VMP1, and TRAIL, previously identified as differentially methylated in IBD compared to controls (2,3). Blood samples were collected from 15 multiplex IBD families in South-Eastern Norway, between April 2023 and January 2024. For comparison, blood samples from 50 sporadic IBD patients and 50 symptomatic controls (SC) were obtained from the IBSEN III cohort (4).
RNA was extracted from PAXgene blood samples of the multiplex families at EpiGen, Akershus University Hospital, Norway, while RNA extractions from the sporadic IBD patients and SCs were performed at The Institute of Clinical Molecular Biology, Kiel, Germany. All cDNA synthesis were carried out at EpiGen. Gene expression was measured using RT-qPCR on cDNA with target-specific primers. Relative quantification was performed using GAPDH and ACTB as housekeeping genes.
Simple t-tests were conducted to compare the mean gene expression between the four comparison groups and across IBD phenotypes.
Results
The multiplex IBD family study included 62 individuals, 41 IBD patients, 25 patients with UC, 16 with CD, and 21 healthy family members, 13 FDR and 8 spouses. The results for the differences in mean fold changes between the four groups, including CD and UC for each of the four candidate genes, are presented in Table 1. The gene expression analysis revealed differences in mean fold changes between the four groups and across CD and UC.
The relative quantification of gene expressions for the four candidate genes revealed no significant differences in gene expression among the IBD patients vs healthy members of multiplex families.
Significant differences were observed for all candidate genes among familial IBD vs sporadic IBD and between healthy multiplex family members vs symptomatic controls, except for BCL3 and TRAIL, respectively. (p<0.005).
| FamIBD vs Fam healthy | FamUC vs Fam healthy | FamCD vs Fam healthy | FamIBD vs sporadic IBD | FamUC vs sporadic UC | FamCD vs sporadic CD | Fam Healthy vs SC |
| Gene | Diff 95% CI | p | Diff 95% CI | p | Diff 95% CI | p | Diff 95% CI | p | Diff 95% CI | p | Diff 95% CI | p | Diff 95% CI | p |
| FKBP5 | 0.13 -0.25-0.5 | 0.501 | 0.07 0.33-0.46 | 0.727 | 0.19 -0.3-0.68 | 0.433 | -0.59 -0.83--0.35 | <0.001
| -0.73 -1.31--0.16 | 0.016 | -0.57 -0.95--0.19 | 0.005 | -0.55 -0.74--0.36 | <0.001 |
| BCL3 | -0.14 -0.4-0.12 | 0.286 | -0.14 -0.4-0.13 | 0,297 | -0.14 -0.43-0.16 | 0.355 | 0.1- 0.06-0.25 | 0.225 | 0.2 -0.03-0.44 | 0.087 | 0.13 -0.14-0.39 | 0.33 | 0.38 0.06-0.69 | 0.021 |
| VMP1 | -0.03 -0.25-0.19 | 0.785 | 0.01 0.25-0.26 | 0,951 | -0.07 -0.32-0.17 | 0.545 | -0.3 -0.44-0.16 | <0.001
| -0.25 -0.48--0.02 | 0.031 | -0.48 -0,73--0.23 | 0.001 | -0.24 -0.41--0.07 | 0.007 |
| TRAIL | 0.03 -0.13-0.19 | 0.716 | 0.02 -0.16-0.2 | 0,821 | 0.04 -0.19-0.27 | 0.724 | -0.23 -0.36--0.09 | 0.001 | -0.21 -0.38--0.04 | 0.018 | -0.3 -0.55--0.04 | 0.023 | -0.04 -0.17-0.09 | 0.578 |
Conclusion
Our results reveal associations between the expression levels of IBD candidate genes and shared family environment, suggesting that epigenetic mechanisms may contribute to the development of familial IBD. Further methylation studies are warranted to clarify the underlying mechanisms of changed gene expression.
References
1. Aabrekk TB. Increased familial risk of IBD over thirty years based on twopulation-based IBD incident cohorts conducted in South-Eastern Norway in 1990-1994 (IBSEN I) and in 2017-2019 (IBSEN III). [Abstract]. In press 2024.
2. Ventham NT, Kennedy NA, Adams AT, Kalla R, Heath S, O'Leary KR, et al. Integrative epigenome-wide analysis demonstrates that DNA methylation may mediate genetic risk in inflammatory bowel disease. Nat Commun. 2016;7:13507.
3. Joustra V, Hageman IL, Satsangi J, Adams A, Ventham NT, de Jonge WJ, et al. Systematic Review and Meta-analysis of Peripheral Blood DNA Methylation Studies in Inflammatory Bowel Disease. J Crohns Colitis. 2023;17(2):185-98.
4. Kristensen VA, Opheim R, Perminow G, Huppertz-Hauss G, Detlie TE, Lund C, et al. Inflammatory bowel disease in South-Eastern Norway III (IBSEN III): a new population-based inception cohort study from South-Eastern Norway. Scand J Gastroenterol. 2021;56(8):899-905.
Disclosure
Vendel A. Kristensen har reported scientific advisory boards from Takeda AS, consultant fees from Janssen-Cilag AS and speaker fees from Thermo Fisher Scientific.
Marte Lie Høivik has reported investigator-initiated research grants from Takeda, Pfizer, Tillotts, Ferring and Janssen. Speaker honoraria from Takeda, Tillotts, Ferring, AbbVie, Pfizer, Galapagos, MSD and Meda and Advisory board Takeda, Galapagos, MSD, Lilly, Janssen, Pfizer and AbbVie.
The other authors of this abstract have no Conflict of Interest