Abstract
Navigation through a dense, physically confining extracellular matrix is common in invasive cell spread and tissue reorganization but is still poorly understood. Here, we show that this migration is mediated by cyclic changes in the activity of a small GTPase RhoA, which is dependent on the oscillatory changes in the activity and abundance of the RhoA guanine nucleotide exchange factor, GEF-H1, and triggered by a persistent increase in the intracellular Ca2+ levels. We show that the molecular clock driving these cyclic changes is mediated by two coupled negative feedback loops, dependent on the microtubule dynamics, with a frequency that can be experimentally modulated based on a predictive mathematical model. We further demonstrate that an increasing frequency of the clock translates into a faster cell migration within physically confining spaces. This work lays the foundation for a better understanding of the molecular mechanisms dynamically driving cell migration in complex environments.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 514-529.e10 |
| Journal | Cell Systems |
| Volume | 13 |
| Issue number | 7 |
| DOIs | |
| State | Published - Jul 20 2022 |
Bibliographical note
Funding Information:The work was supported by NIH grants CA155758-05 (to S.H.L. A.H. H.C. and A.L.), P01CA254849 and U54CA210190 (to J.C.H. and D.J.O.), and U54CA209992 (to S.H.L. A.H. M.S.Y. V.A. H.C. M.M. and A.L.); we thank Andrew Ewald and Alexander Popel for their helpful comments. S.H.L. and A.L. conceived the project. S.H.L. designed and used the 3D collective migration device, performed all the experiments unless otherwise indicated, and analyzed the data. A.H. analyzed the fluorescence resonance energy transfer (FRET) data. M.S.Y. performed the traction force experiment and analysis of the data. V.A. and M.S.Y. fabricated the devices for the analysis of cell migration in spatially confining channels. H.C. constructed the plasmid for detecting Ca2+ activity (GCaMP5). S.H.L. and A.L. performed the computational modeling and analysis. J.C.H. and D.J.O. performed the cell migration simulation modeling, analyzed the data, and wrote the cell migration simulation part in STAR Methods. S.H.L. and A.L. wrote and edited the manuscript. D.J.O. M.M. and A.L. obtained the funding. A.L. supervised the project. The authors declare no competing interests.
Funding Information:
The work was supported by NIH grants CA155758-05 (to S.H.L., A.H., H.C., and A.L.), P01CA254849 and U54CA210190 (to J.C.H. and D.J.O.), and U54CA209992 (to S.H.L., A.H., M.S.Y., V.A., H.C., M.M., and A.L.); we thank Andrew Ewald and Alexander Popel for their helpful comments.
Publisher Copyright:
© 2022 Elsevier Inc.
Keywords
- GEF-H1
- RhoA guanine exchange factor
- cell migration
- computational model
- confining spaces
- intracellular calcium
- microtubule dynamics
- molecular clock
- negative feedback
- oscillations