Z26 - Autologous MPS model of the gut-liver axis and single-cell profiling reveal distinct roles of tissue-resident immune environments in systemic inflammation

Introduction: : The human gut-liver axis represents a critical immunological interface, integrating microbial, dietary, and endogenous signals to maintain homeostasis. This axis is central to immune responses, with the colon and liver containing a significant fraction of the body's T lymphocytes, including tissue-resident memory T cells (Trm). Dysregulation of this axis, as seen in Inflammatory Bowel Disease (IBD), can lead to systemic inflammation. However, understanding of the gut-liver-blood T cell interactions within individuals is limited.

Materials and

Methods: : This study addresses this gap using a novel human multi-organ microphysiological system (MPS) that models the autologous gut-liver axis and their resident immune environments. We combine this MPS with whole-cell and TCR single-cell RNA sequencing (scRNA-seq) of donor-matched large intestine (intraepithelial and lamina propria), liver, and blood, to profile T cell heterogeneity and clonal relationships. This multi-tissue approach, allows us to directly compare T cell phenotypes and states, minimizing inter-individual variability that confounds typical scRNA-seq studies of immune atlases. We systematically challenged our MPS with diverse stimuli (LPS, Poly:IC, 5-OP-RU) to induce controlled inflammatory responses.

Results, Conclusions, and Discussions:: By coupling transcriptional landscapes with clonal architecture, we distinguished tissue-resident subsets from circulating cells and identified how specific niches shape the T cell landscape. The results revealed significant differences in transcriptional profiles, and response dynamics of T cells across the gut-liver-blood axis. Clonal expansion analysis highlights immunodominant responses and tissue-specific adaptations, revealing unique signatures within each compartment that contribute to the nuanced balance between immune surveillance and tolerance.
Our findings provide a new resolution into the autologous tissue-specific immune dynamics of the gut-liver axis. This network-level view of inter-organ inflammatory cascades identifies how gut and liver resident immune cells influence systemic immunity. This study provides a bridge between single-cell profiling and mechanistic understanding of tissue-immune dynamics, with implications for targeted therapies in IBD and other inflammatory conditions. The autologous MPS model, coupled with detailed single-cell analysis, offers a powerful platform for dissecting complex immune interactions and informing interventions that leverage tissue-resident T cell properties.

Acknowledgements and/or References (Optional):: Single-cell sequencing was performed at the JHU Single Cell and Transcriptomics Core. The study was supported by grants from NIGMS 5R35GM146900. R. R. and D. K. B are supported by an award from the Good Ventures Foundation and Open Philanthropy, as well as start-up funds from Case Western Reserve University and University Hospitals.