Human regulatory T-cells locally differentiate and are functionally heterogeneous within the inflamed arthritic joint

Lutter, L.; van der Wal, M. M.; Brand, E. C.; Maschmeyer, P.; Vastert, B.; Mashreghi, M.-F.; van Loosdregt, J.; van Wijk, F.

doi:10.1101/2022.02.18.480998

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bioRxiv · 10.1101/2022.02.18.480998

Human regulatory T-cells locally differentiate and are functionally heterogeneous within the inflamed arthritic joint

Lutter, L.·van der Wal, M. M.·Brand, E. C.·Maschmeyer, P.·Vastert, B.·Mashreghi, M.-F.·van Loosdregt, J.·van Wijk, F.

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Abstract

ObjectiveTregs are crucial for immune regulation, and environment-driven adaptation of effector (e)Tregs is essential for local functioning. However, the extent of human Treg heterogeneity in inflammatory settings is unclear. MethodsWe combined single-cell RNA- and TCR-sequencing on Tregs derived from 4-6 patients with juvenile idiopathic arthritis (JIA) to investigate the functional heterogeneity of human synovial fluid (SF)-derived Tregs from inflamed joints. Confirmation and suppressive function of the identified Treg clusters was assessed by flow cytometry. ResultsFour Treg clusters were identified; incoming, activated eTregs with either a dominant suppressive or cytotoxic profile, and GPR56+CD161+CXCL13+ Tregs. Pseudotime analysis showed differentiation towards either classical eTreg profiles or GPR56+CD161+CXCL13+ Tregs supported by TCR data. Despite its most differentiated phenotype GPR56+CD161+CXCL13+ Tregs were shown to be suppressive. Furthermore, BATF was identified as an overarching eTreg regulator, with the novel Treg-associated regulon BHLHE40 driving differentiation towards GPR56+CD161+CXCL13+ Tregs, and JAZF1 towards classical eTregs. ConclusionOur study reveals a heterogeneous population of Tregs at the site of inflammation in JIA. SF Treg differentiate to a classical eTreg profile with a more dominant suppressive or cytotoxic profile that share a similar TCR repertoire, or towards GPR56+CD161+CXCL13+ Tregs with a more distinct TCR repertoire. Genes characterizing GPR56+CD161+CXCL13+ Tregs were also mirrored in other T-cell subsets in both the tumor and autoimmune setting. Finally, the identified key regulators driving SF Treg adaptation may be interesting targets for autoimmunity or tumor interventions.

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Lutter, L., van der Wal, M. M., Brand, E. C., Maschmeyer, P., Vastert, B., Mashreghi, M.-F., van Loosdregt, J., van Wijk, F.. 2022-02-20. Human regulatory T-cells locally differentiate and are functionally heterogeneous within the inflamed arthritic joint. https://doi.org/10.1101/2022.02.18.480998

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