High-Throughput Mechanomic Screening Reveals Novel Regulators of Single-Cell Mechanics
Laura von Selzam,
Katarzyna Plak,
Christine Schweitzer,
Cornelia Liebers,
Paul Müller,
Marta Urbanska,
Martin Kräter,
Buzz Baum,
Jona Kayser, et al.
Biophysical Journal
125
(12)
3060-3073
(2026)
| Journal
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The mechanical properties of cells are dynamic, allowing them to adjust to different needs in different biological contexts. In recent years, advanced biophysical techniques have enabled the rapid, high-throughput assessment of single-cell mechanics, providing new insights into the regulation of the mechanical cell phenotype. However, the molecular mechanisms by which cells maintain and regulate their mechanical properties remain poorly understood. Here, we present a genome-scale RNA interference (RNAi) screen investigating the roles of kinase and phosphatase genes in regulating single-cell mechanics using Real-Time Fluorescence and Deformability Cytometry (RT-FDC). Our screen identified 82 known and novel mechanical regulators across diverse cellular functions from 214 targeted genes, leveraging RT-FDC’s unique capabilities for comprehensive, high-throughput mechanical phenotyping with single-cell and cell cycle resolution. These findings refine our understanding of how signaling pathways coordinate structural determinants of cell mechanical phenotypes and provide a starting point for uncovering new molecular targets involved in biomechanical regulation across diverse biological systems.
Spatial resource dynamics control resistance escape
Nico Appold,
Timon Citak,
Auguste Palm,
Jona Kayser
The evolution of therapy resistance in structured populations such as biofilms and solid tumours is shaped by emergent spatial organization, with pro-found consequences for evolution-based therapies. However, how treatment reshapes these patterns remains poorly understood. Here we show that intermittent treatment pulses transiently reconfigure the resource landscape, reorganize spatial growth zones, and can enable resistant mutants to escape spatial confinement and drive therapy failure. We introduce a spatial evolution assay in which populations expand from single, genetically tailored yeast cells, enabling quantitative tracking of the full spatiotemporal trajectories of continually emerging resistant mutants under intermittent treatment. By integrating these observations with a mechanistically interpretable \textit{in silico} model in a real-to-sim-to-real loop, we identify an effective phase transition in schedule space that defines an optimal balance between population control and sustained resistance confinement, which we validate experimentally. Together, our results establish resource-mediated spatial confinement as a central organizing principle of resistance evolution and provide a mechanistic foundation for spatially informed, evolution-based therapies.
Kontakt
Forschungsgruppe Jona Kayser
Max-Planck-Institut für die Physik des Lichts Staudtstr. 2 91058 Erlangen
