Project description:To investigate the mechanisms of PI3Kα-induced senescence, we performed a gene expression microarray analysis with MCF-10A/H and parental MCF-10A cells.
Project description:We set out to characterize the transcriptional heterogeneity of the senescence program using a large number of whole-transcriptome sequencing datasets generated by us or publicly available. We identify a number of senescence transcriptional signatures associated to specific stresses or cell types. We also merge all the studies to identify and validate the genes that are universally differentially regulated during senescence.
Project description:Cells are subjected to dynamic mechanical environments which impart various forces and induce cellular responses. In age-related conditions like pulmonary fibrosis, there is both an increase in tissue stiffness and an accumulation of senescent cells, leading to elevated tension in fibroblasts among other cells. While senescent cells produce a senescence-associated secretory phenotype (SASP), the impact of physical stimuli on both cellular senescence and SASP is not well understood. Here, we show that mechanical tension, modeled using cell culture substrate rigidity, influences senescent cell markers like SA-beta-gal and secretory phenotypes. Comparing human primary pulmonary fibroblasts (IMR-90) cultured on physiological (2 kPa), fibrotic (50 kPa), and plastic (3 GPa) substrates followed by senescence induction using doxorubicin, we identified unique high-stiffness-driven secretory protein profiles using mass spectrometry and transcriptomic signatures, both showing an enrichment in collagen proteins. Computational meta-analysis of human interstitial lung disease single-cell RNA sequencing datasets confirmed these genes are highly expressed in disease samples and strongly correlate with mechanotransduction and senescence-related pathways. Thus, mechanical forces shape cell senescence and their secretory phenotypes.
Project description:Cells are subjected to dynamic mechanical environments which impart forces and induce cellular responses. In age-related conditions like pulmonary fibrosis, there is both an increase in tissue stiffness and an accumulation of senescent cells. While senescent cells produce a senescence-associated secretory phenotype (SASP), the impact of physical stimuli on both cellular senescence and the SASP is not well understood. Here, we show that mechanical tension, modeled using cell culture substrate rigidity, influences senescent cell markers like SA-β-gal and secretory phenotypes. Comparing human primary pulmonary fibroblasts (IMR-90) cultured on physiological (2 kPa), fibrotic (50 kPa), and plastic (approximately 3 GPa) substrates, followed by senescence induction using doxorubicin, we identified unique high-stiffness-driven secretory protein profiles using mass spectrometry and transcriptomic signatures, both showing an enrichment in collagen proteins. Consistently, clusters of p21+ cells are seen in fibrotic regions of bleomycin induced pulmonary fibrosis in mice. Computational meta-analysis of single-cell RNA sequencing datasets from human interstitial lung disease confirmed these stiffness SASP genes are highly expressed in disease fibroblasts and strongly correlate with mechanotransduction and senescence-related pathways. Thus, mechanical forces shape cell senescence and their secretory phenotypes.
Project description:Neutrophils are the most abundant leukocytes in human peripheral blood but exhibit considerable heterogeneity, with distinct subsets characterized by diverse functional states. Lipopolysaccharide (LPS) has been shown to drive neutrophil senescence, a process distinct from apoptosis. However, the global transcriptional landscape of LPS-induced aged neutrophils remains largely unexplored. Here, we performed whole-transcriptome RNA sequencing (RNA-seq) on primary human neutrophils isolated from healthy donors. Neutrophils were treated with LPS to induce senescence, with untreated cells serving as controls. Our comparative transcriptomic analysis revealed that senescent neutrophils display significant upregulation of multiple genes associated with neutrophil extracellular trap (NET) formation compared to non-aged neutrophils. This dataset provides a comprehensive resource for investigating the molecular signatures underlying neutrophil senescence and its potential implications in inflammation and host defense.