Chesla, D. | Search bioRxiv

bioRxiv2025

Comparative immuno-biology at clinical recognition of early multiple organ dysfunction syndrome in pediatric and adult patients using single-cell transcriptomics

Globally, sepsis remains a major health issue, with Multiple Organ Dysfunction Syndrome (MODS) being a leading cause of mortality. MODS, a severe condition often seen in intensive care units, typically results from infections or trauma and involves complex pathophysiological processes requiring various clinical interventions. Although infections are the main triggers, the mechanisms driving MODS remain unclear. To investigate the transition of sepsis to MODS, we generated a single cell RNA sequencing dataset comprising 86,839 immune cells from pediatric sepsis patients at the clinical onset of MODS patients and age-matched controls, identifying 22 distinct cell types. A cluster of S100 genes, located in the same genomic region, was highly expressed in neutrophils in MODS patients, demonstrating strong diagnostic potential across cohorts (AUC=0.94- 0.99) and potential as therapeutic targets. We found that many B and T cells showed heightened inflammation and increased apoptotic activity during early MODS. Additionally, specific transcription regulators and surface proteins associated with inflammation and S100 regulations were uniquely expressed in MODS. Pseudotime analysis revealed distinct S100 gene expression patterns between controls and MODS. Cell-cell interaction analysis highlighted dendritic cells as key mediators, enhancing communication between plasma cells and V{delta} T cells while activating inflammatory and immunosuppressive pathways. We also analyzed 116,803 immune cells from adult MODS patients, revealing stronger immune dysregulation compared to pediatric MODS, including altered S100 gene expression, and enhanced cell-cell interactions. These findings suggest that S100 genes may serve as a marker for MODS. Furthermore, insights gained from adult MODS could improve our understanding of rare pediatric MODS and contribute to the development of better therapeutics for all MODS patients.

bioinformatics↗

bioRxiv2023

Bulk RNA-Sequencing of small airway cell cultures from IPF and post-COVID lung fibrosis patients illustrates disease signatures and differential responses to TGF-β1 treatment

IPF is a condition in which an injury to the lung leads to the accumulation of scar tissue. This fibrotic tissue reduces lung compliance and impairs gas exchange. Studies have shown that infection with COVID-19 significantly worsens the clinical outcomes of IPF patients. The exact etiology of IPF is unknown, but recent evidence suggests that the distal small airways, (those having a diameter less than 2 mm in adults), play a role in the early pathogenesis of IPF. TGF-{beta}1 is a main driver of fibrosis in a variety of tissues; the binding of TGF-{beta}1 to its receptor triggers a signaling cascade that results in inflammatory signaling, accumulation of collagen and other components of the extracellular matrix, and immune system activation. This study aimed to investigate possible mechanisms that contribute to worsening lung fibrosis in IPF patients after being diagnosed with COVID-19, with a particular focus on the role of TGF-{beta}1. Small airway cell cultures derived from IPF and post-COVID-19 IPF patient transplant tissues were submitted for RNA-sequencing and differential gene expression analysis. The genetic signatures for each disease state were determined by comparing the differentially expressed genes present in the cells cultured under control conditions to cells cultured with TGF-{beta}1. The genes shared between the culture conditions laid the framework for determining the genetic signatures of each disease. Our data found that genes associated with pulmonary fibrosis appeared to be more highly expressed in the post-COVID fibrosis samples, under both control and TGF-{beta}1-treated conditions. A similar trend was noted for genes involved in the TGF-{beta}1 signaling pathway; the post-COVID fibrosis cell cultures seemed to be more responsive to treatment with TGF-{beta}1. Gene expression analysis, RT-PCR, and immunohistochemistry confirmed increased levels of BMP signaling in the IPF small airway cell cultures. These findings suggest that TGF-{beta}1 signaling in IPF small airway cells could be inhibited by BMP signaling, leading to the differences in genetic signatures between IPF and post-COVID fibrosis.

cell biology↗