Biology subjects

Basualto-Alarcon, C.

Publications and source records attributed to Basualto-Alarcon, C..

3 recordsLinked to original sources

bioRxiv2025

High-content live-cell time-lapse imaging predicts cells about to die via apoptosis

Cell death is a dynamic process that unfolds through time. Live-cell time-lapse imaging captures these dynamics in a way thats impossible for static snapshots. High-content imaging (HCI), which has been developed for static microscopy, applied to time-lapse imaging can quantify how single-cell states change through time. Here we show the ability of high-content live-cell time-lapse imaging (HCLTI) to quantify the onset and progression of one form of cell death called apoptosis. We apply the Live Cell Painting assay called ChromaLIVETM and develop an HCLTI analysis pipeline. We show that HCLTI can discern the morphology dynamics of cells undergoing apoptosis, and demonstrate that machine learning can predict apoptosis as early as 100 minutes after exposing HeLa cells to the apoptosis inducer Staurosporine. This technical advancement paves the way for future studies to better understand the dynamics of other forms of cell death. Understanding cell death dynamics is one piece of solving larger biomedical puzzles like understanding how cells resist death (e.g., therapeutic resistance of cancer cells) and how cells die too soon (e.g., neurodegeneration).

systems biology↗

bioRxiv2024

A morphology and secretome map of pyroptosis

Pyroptosis represents one type of Programmed Cell Death (PCD). It is a form of inflammatory cell death that is canonically defined by caspase-1 cleavage and Gasdermin-mediated membrane pore formation. Caspase-1 initiates the inflammatory response (through IL-1{beta} processing), and the N-terminal cleaved fragment of Gasdermin D polymerizes at the cell periphery forming pores to secrete pro-inflammatory markers. Cell morphology also changes in pyroptosis, with nuclear condensation and membrane rupture. However, recent research challenges canon, revealing a more complex secretome and morphological response in pyroptosis, including overlapping molecular characterization with other forms of cell death, such as apoptosis. Here, we take a multimodal, systems biology approach to characterize pyroptosis. We treated human Peripheral Blood Mononuclear Cells (PBMCs) with 36 different combinations of stimuli to induce pyroptosis or apoptosis. We applied both secretome profiling (nELISA) and high-content fluorescence microscopy (Cell Painting). To differentiate apoptotic, pyroptotic and control cells, we used canonical secretome markers and modified our Cell Painting assay to mark the N-terminus of Gasdermin-D. We trained hundreds of machine learning (ML) models to reveal intricate morphology signatures of pyroptosis that implicate changes across many different organelles and predict levels of many pro-inflammatory markers. Overall, our analysis provides a detailed map of pyroptosis which includes overlapping and distinct connections with apoptosis revealed through a mechanistic link between cell morphology and cell secretome.

systems biology↗

bioRxiv2018

Acute Psychological Stress Triggers Circulating Cell-Free Mitochondrial DNA

Intrinsic biological mechanisms transduce psychological stress into physiological adaptation, but the role of mitochondria and mitochondrial DNA (mtDNA) in this process has not been defined in humans. Here, we show that similar to physical injury, psychological stress triggers elevation in circulating cell- free mtDNA (ccf-mtDNA). Healthy midlife adults exposed on two separate occasions to a brief psychological challenge exhibit a 2-3-fold increase in ccf-mtDNA, with no change in nuclear DNA levels, establishing the magnitude and specificity to ccf-mtDNA. In cell-based studies, we show that glucocorticoid signaling - a consequence of psychological stress in humans - is sufficient to induce mtDNA extrusion in a time frame consistent with human psychophysiology. Collectively, these findings provide the first evidence that psychological stress induces ccf-mtDNA and implicate glucocorticoid signaling as a trigger for ccf-mtDNA release. Further work is needed to examine the functional significance of psychological stress-induced ccf-mtDNA as a mitokine in humans.

molecular biology↗