Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
[Preprint]. 2024 Dec 31:2024.12.30.24319783.
doi: 10.1101/2024.12.30.24319783.

Multi-omic Biomarkers Distinguish Rheumatoid Arthritis in Discordant Monozygotic Twins

Rebecca B Blank  1 , Kevin Bu  2 , Weixi Chen  3 , Ian Cunningham  1 , Jeremy Sokolove  4 , Lauren Lahey  4 , Adriana Heguy  5 , Rhina Medina  1 , Carles Ubeda  1   2   3   4   5   6   7 , Renuka R Nayak  6 , Jiyuan Hu  3 , Adam Cantor  2 , Jakleen Lee  2 , Jose C Clemente  2 , Jose U Scher  1   7
Affiliations

Multi-omic Biomarkers Distinguish Rheumatoid Arthritis in Discordant Monozygotic Twins

Rebecca B Blank et al. medRxiv. .

Update in

Abstract

Background: Although genetic factors have been identified in the pathogenesis of rheumatoid arthritis (RA), the concordance rate in monozygotic (MZ) twins is low, suggesting that other features contribute to disease development. Further, the relative contribution of such non-genetic elements in identical twins have not been characterized. Here, we aimed to measure differentiating host and microbial biomarkers of RA by studying MZ twins discordant for disease using a multi-omics approach.

Methods: Eight pairs of MZ twins discordant for RA (n=16) were enrolled. Gut microbiome was assessed using shotgun metagenomic sequencing. Autoantibodies, cytokines, and other plasma proteins were measured in both plasma and feces. Levels of short and medium-chain fatty acids from serum and feces were quantified using gas chromatography mass spectrometry (GC-MS).

Results: While overall microbiome diversity and composition did not significantly differ between twins, we observed a decrease in Blautia faecis in affected twins. Affected twins had higher concentrations of both fecal and plasma citrullinated and non-citrullinated autoantibodies, as well as significantly lower concentrations of fecal butyrate and propionate.

Conclusion: Multi-omics biomarkers differentiate MZ twins discordant for RA. Blautia faecis, which is associated with reduced inflammatory cytokine expression, was decreased in RA twins. Similarly, short-chain fatty acids, known to have immune modulatory effects, were decreased in affected twins, suggesting further bi-directional interactions between inflammation at the gut barrier and disease state. If confirmed in other cohorts, exhaustive multi-omics approaches may improve our understanding of RA pathogenesis and potentially contribute to novel diagnostics and co-adjuvant therapies.

PubMed Disclaimer

Conflict of interest statement

Competing interests: RBB, KB, WC, IC, AH, RH, RRN, JH, AC, JL, JS, LL, CU, JCC, have no conflicts to declare. JUS has served as a consultant for Janssen, Novartis, Pfizer, Sanofi, Amgen, UCB, BMS, and AbbVie; and has received funding for investigator-initiated studies from Janssen and Pfizer.

Figures

Figure 1.
Figure 1.
Gut bacterial composition and predicted functional pathways in RA and unaffected twins. (A) Shannon alpha diversity in participants, with box plots representing median and interquartile range. Lines are drawn between MZ pairs. (B) Principal coordinate analysis (PCoA) of Bray-Curtis beta diversity between groups. (C) Linear discriminant analysis Effect Size (LEfSe) showing differential species in RA twins compared to unaffected twins. (D) Boxplot showing distribution of B. faecis abundance in unaffected and RA twins. (E) Barplot of Wilcoxon effect sizes for differential pathways. *p<0.05, **p<0.01.
Figure 2.
Figure 2.
Comparison of fecal short chain fatty acid (SCFA) concentrations between unaffected participants and RA twins. (A) RA twins have lower fecal concentration of acetate. (B) Butyrate fecal levels are depleted in RA twins. (C) Propionate fecal concentration is lower in affected twins. (D) Valerate fecal levels are higher in RA twins. Lines are drawn between MZ pairs. P-values calculated with paired t test; *p<0.05.
Figure 3.
Figure 3.
Comparison of citrullinated and non-citrullinated fecal and plasma autoantibody concentration between unaffected and RA twins as measured by mean fluorescence intensity (MFI). Fecal autoantibody boxplots (A) and plasma autoantibody boxplots (B) are shown. Box plots represent median and interquartile range. Lines are drawn between MZ pairs. P-values calculated with paired t test; *p<0.05.
Figure 3.
Figure 3.
Comparison of citrullinated and non-citrullinated fecal and plasma autoantibody concentration between unaffected and RA twins as measured by mean fluorescence intensity (MFI). Fecal autoantibody boxplots (A) and plasma autoantibody boxplots (B) are shown. Box plots represent median and interquartile range. Lines are drawn between MZ pairs. P-values calculated with paired t test; *p<0.05.
Figure 4.
Figure 4.
Levels of circulating cytokines and other proteins in RA and unaffected twins. Proximity extension analysis of 92 inflammation-related plasma proteins revealed significant elevation of (A) IL-1α, (B) CXCL1, (C) CCL23, (D) CCL25, and (E) MMP-10 in RA versus unaffected twin. Y axes represent normalized expression values (NPX) in log2 scale. Box plots represent median and interquartile range. Lines are drawn between MZ pairs. P-values calculated with paired t test; *p<0.05, **p<0.01.
Figure 5.
Figure 5.
Cohen’s D (paired) effect sizes for top differential features of each datatype along with directionality of enrichment.
See this image and copyright information in PMC

References

    1. Silman AJ, MacGregor Aj Fau - Thomson W, Thomson W Fau - Holligan S, et al. Twin concordance rates for rheumatoid arthritis: results from a nationwide study. Br J Rheumatol 1993;32(10):903–7. (In eng). DOI: 10.1093/rheumatology/32.10.903. - DOI - PubMed
    1. Aho K, Koskenvuo M, Tuominen J, Kaprio J. Occurrence of rheumatoid arthritis in a nationwide series of twins. J Rheumatol 1986;13(5):899–902. (https://www.ncbi.nlm.nih.gov/pubmed/3820198). - PubMed
    1. Maeda Y, Kurakawa T, Umemoto E, et al. Dysbiosis Contributes to Arthritis Development via Activation of Autoreactive T Cells in the Intestine. Arthritis Rheumatol 2016;68(11):2646–2661. DOI: 10.1002/art.39783. - DOI - PubMed
    1. Wu HJ, Ivanov II, Darce J, et al. Gut-residing segmented filamentous bacteria drive autoimmune arthritis via T helper 17 cells. Immunity 2010;32(6):815–27. DOI: 10.1016/j.immuni.2010.06.001. - DOI - PMC - PubMed
    1. Nayak RR, Alexander M, Deshpande I, et al. Methotrexate impacts conserved pathways in diverse human gut bacteria leading to decreased host immune activation. Cell Host & Microbe 2021;29(3):362–377.e11. DOI: 10.1016/j.chom.2020.12.008. - DOI - PMC - PubMed

Publication types

LinkOut - more resources