PT  - JOURNAL ARTICLE
AU  - Butera, Francesca
AU  - Sero, Julia E.
AU  - Dent, Lucas
AU  - Bakal, Chris
TI  - Actin networks modulate heterogenous NF-κB dynamics in response to TNFα
AID  - 10.1101/2022.01.19.476961
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2022.01.19.476961
4099  - http://biorxiv.org/content/early/2022/01/21/2022.01.19.476961.short
4100  - http://biorxiv.org/content/early/2022/01/21/2022.01.19.476961.full
AB  - The canonical NF-κB transcription factor RELA is a master regulator of immune and stress responses and is commonly upregulated in PDAC tumours. Using live imaging, we characterised single cell RELA translocation dynamics in two human PDAC cell lines and identified high cell-to-cell variability in RELA responses to TNFα, including unresponsive, damped, and sustained nuclear RELA localisation. Using a combination of quantitative single cell imaging and Bayesian analysis, we determined that heterogeneity in RELA nuclear translocation between and within PDAC cell lines is dependent on cytoskeletal organisation, in particular actin abundance and distribution. Subsequently, RELA nuclear localisation could be up or downregulated through biochemical modulation of cell shape and the cytoskeleton, particularly by disrupting nucleation of actin stress fibres and branched actin via formin and ARP2/3 inhibition. Together, our data provide evidence that actin configuration regulates RELA translocation during the inflammatory response and that targeting actin dynamics can be used to modulate misregulated NF-κB signalling in PDAC.Competing Interest StatementThe authors have declared no competing interest.