Investigation of dynamic regulation of TFEB nuclear shuttling by microfluidics and quantitative modelling.

Communications biology 2025

Transcription Factor EB (TFEB) controls lysosomal biogenesis and autophagy in response to nutritional status and other stress factors. Although its regulation by nuclear translocation is known to involve a complex network of well-studied regulatory processes, the precise contribution of each of these mechanisms is unclear. Using microfluidics technology and real-time imaging coupled with mathematical modelling, we explored the dynamic regulation of TFEB under different conditions. We found that TFEB nuclear translocation upon nutrient deprivation happens in two phases: a fast one characterised by a transient boost in TFEB dephosphorylation dependent on transient calcium release mediated by mucolipin 1 (MCOLN1) followed by activation of the Calcineurin phosphatase, and a slower one driven by inhibition of mTORC1-dependent phosphorylation of TFEB. Upon refeeding, TFEB cytoplasmic relocalisation kinetics are determined by Exportin 1 (XPO1). Collectively, our results show how different mechanisms interact to regulate TFEB activation and the power of microfluidics and quantitative modelling to elucidate complex biological mechanisms.

4 Figures Extracted

Fig. 1 PMC

Experimental measurements of TFEB nuclear shuttling dynamics in response to mTOR inhibition and refeeding in single cells by microfluidics and microsc...

Fig. 2 PMC

Experimental measurements of the nucleocytoplasmic shuttling of TFEB following mTOR inhibition: features representation. A Starvation ‘step’ experime...

Fig. 3 PMC

Quantitative model of TFEB nuclear shuttling dynamics. A Exportin 1 inhibition ‘step’ experiment: HeLa cells were kept for 180 min in RPMI, then swit...

Fig. 4 PMC

Experimental measurements and modelling of TFEB nuclear shuttling dynamics following Calcineurin and Calcium perturbations. A Thapsigargin ‘step’ exp...