Project description:Langerhans cells (LC) in skin help initiate the immune response to locally presented antigens. We performed high-resolution single-cell RNA-sequencing (scRNAseq) analysis for antigen presenting cells including LC in normal mouse skin, and in mouse skin expressing the human papillomavirus (HPV) 16 E7 oncogene. Ear skin was collected from normal and trangenic mice. Dissociated CD45+ cells were processed for scRNA-seq using the 10X Genomics Chromium 3' gene expression kit (v2).
Project description:Kilian2024 - Immune cell dynamics in Cue-Induced Extended Human Colitis Model
Single-cell technologies such as scRNA-seq and flow cytometry provide critical insights into immune cell behavior in inflammatory bowel disease (IBD). However, integrating these datasets into computational models for dynamic analysis remains challenging. Here, Kilian et al., (2024) developed a deterministic ODE-based model that incorporates these technologies to study immune cell population changes in murine colitis. The model parameters were optimized to fit experimental data, ensuring an accurate representation of immune cell behavior over time. It was then validated by comparing simulations with experimental data using Pearson’s correlation and further tested on independent datasets to confirm its robustness. Additionally, the model was applied to clinical bulk RNA-seq data from human IBD patients, providing valuable insights into immune system dynamics and potential therapeutic strategies.
Figure 4c, obtained from the simulation of human colitis model is highlighted here.
This model is described in the article:
Kilian, C., Ulrich, H., Zouboulis, V.A. et al. Longitudinal single-cell data informs deterministic modelling of inflammatory bowel disease. npj Syst Biol Appl 10, 69 (2024). https://doi.org/10.1038/s41540-024-00395-9
Abstract:
Single-cell-based methods such as flow cytometry or single-cell mRNA sequencing (scRNA-seq) allow deep molecular and cellular profiling of immunological processes. Despite their high throughput, however, these measurements represent only a snapshot in time. Here, we explore how longitudinal single-cell-based datasets can be used for deterministic ordinary differential equation (ODE)-based modelling to mechanistically describe immune dynamics. We derived longitudinal changes in cell numbers of colonic cell types during inflammatory bowel disease (IBD) from flow cytometry and scRNA-seq data of murine colitis using ODE-based models. Our mathematical model generalised well across different protocols and experimental techniques, and we hypothesised that the estimated model parameters reflect biological processes. We validated this prediction of cellular turnover rates with KI-67 staining and with gene expression information from the scRNA-seq data not used for model fitting. Finally, we tested the translational relevance of the mathematical model by deconvolution of longitudinal bulk mRNA-sequencing data from a cohort of human IBD patients treated with olamkicept. We found that neutrophil depletion may contribute to IBD patients entering remission. The predictive power of IBD deterministic modelling highlights its potential to advance our understanding of immune dynamics in health and disease.
This model was curated during the Hackathon hosted by BioMed X GmbH in 2024.
Project description:<p>The role of sebaceous glands (SGs) in the early pathogenesis of hidradenitis suppurativa (HS) is undefined, with unclear causal relationship to inflammatory sequelae. The aim of this study was to determine whether SG aberrations constitute a primary pathogenic driver in early HS and elucidate the underlying mechanism. Histological study, single-cell RNA sequencing (scRNA-seq), and in vitro functional assays were employed. Clinical specimens included non-lesional skin, early lesional skin from patients diagnosed with Hurley I HS, and healthy controls. Human SZ95 sebocytes were used for mechanistic studies, including gene knockdown with lentivirus, bulk RNA sequencing, and liquid chromatography-tandem mass spectrometry based lipidomic analysis. </p><p>The size of SG was significantly reduced in HS patients. Development aberration and significant downregulation of tight junction signaling (e.g., TJP1, OCLN, CLDN1) in HS SGs were revealed by scRNA-seq, associated with compromised barrier integrity and early CD45+ immune cell infiltration. Knockdown of CLDN1 in human SZ95 sebocytes recapitulated these findings, inducing a robust pro-inflammatory response and a shift toward keratinocyte-like lineage accompanied by metabolic reprogramming, specifically overproduction of lysophosphatidylcholine (LPC). Exogenous LPC directly promoted proliferation and inflammatory cytokine secretion in HaCaT keratinocytes. Collectively, these findings reposition SGs as instigators in the early pathogenesis of HS.</p>
Project description:Liver has abundant regenerative capacity, but hepatocytes cannot be cultured long-term in vitro. Activation of FXR signaling induced iHOs with both hepatocyte traits and proliferative potential. To investigate the transcriptomic analysis of iHOs, we performed bulk RNA-seq analysis of organoids in different passages and single cell RNA-seq analysis of iHOs. Compared to 2D-iPS-Heps and organoids in other culture conditions, iHOs have expression profiles more similar to primary human hepatocytes and human fetal liver cells. Single cell RNA-seq revealed iHOs have a fraction that co-expresses hepatocyte markers and proliferation markers.