Optogenetic stimulation of Lbc GEF-mediated Rho activity dynamics promotes cell invasion
Posted on: 25 April 2025 , updated on: 29 April 2025
Preprint posted on 31 March 2025
Categories: cancer biology, cell biology
Background and hypothesis.
Directional cell migration is a multi-step process involving under the control of biochemical and mechanical signal cues and is vital for development and homeostasis of multicellular organisms. Failure at any step causes aberrant migration behaviour, as observed in cancer cell metastasis, where tumor cells invade surrounding tissues and migrate to distant sites, and this process is dependent on actin-driven protrusions and contractile forces.
Rho GTPases are key regulators of cell contraction, driving myosin-based contraction pulses. RhoA activity is controlled by an upstream mechanosensitive signalling network that include Lbc-type guanine nucleotide exchange factors (GEFs) like GEF-H1. Further, RhoA activity is under a fast-acting positive feedback loop that intensifies its activity and delays the negative feedback that ultimately suppresses it.
Highly invasive cancers, including breast and melanoma, have elevated levels of GEF-H1, indicating a potential link between Rho-driven contraction dynamics and tumor invasiveness. To investigate this link, the authors of this preprint established an optogenetic tumor spheroid model, offering precise light-based control of GEF-H1 activity and, subsequently, Rho-mediated contraction dynamics. The authors hypothesized that increased contraction dynamics, induced by optogenetic activation of GEF-H1 and following Rho signalling, generate mechanical forces asymmetry promoting tumor spheroid expansion and facilitate the escape of invasive cancer cells.
Key findings with an analogy.
A Busy Train Station with Escalators.
Imagine a chaotic train station during peak hours. The station is jam-packed with commuters (here: cancer cells) looking to escape out into the city (here: invading surrounding tissues). Now, people can’t just leave on their own—they’re directed by automatic escalators and moving walkways, comparable to how cells are directed by protrusions and contraction. These pathways are active only when certain control systems are triggered.
Figure 1. (Reproduced from Figure S3). Multicellular melanoma spheroids (3D cell culture) expand upon optogenetic stimulation of contraction dynamics. Bottom panels show a proposed mechanism that could explain the experimental observations.
Finding 1: The Escalator Controller (GEF-H1)
The control room has a station manager—let’s call her GEF-H1. She can control when and where (+ in which direction) escalators begin to move. If she is super active, she starts turning on escalators and moving walkways all over the station. This increases the traffic of people—some commuters suddenly find themselves being directed toward the edges of the station faster than before. In the preprint, the authors demonstrate—using 3D multicellular tumor spheroids—that light-controlled GEF-H1 activation boosted single-cell motility and triggered the entire spheroid to expand.
Finding 2: Foot Sensors (FAK Adhesions)
But people are not simply sliding around – they step on pressure-sensitive tiles (here: FAK, the focal adhesion kinase) that can decide if somebody is ready to move forward or not. These tiles can sense pressure and anchor people, so they don’t slip, providing them with adequate grasp to push off and move ahead. So, when the station manager, GEF-H1, turns up the escalator speed and the foot sensors are activated, people can now exit the station more efficiently, even if they weren’t planning to. In the preprinted study, the authors observed expansion of spheroids, dependent on FAK activity, as FAK inhibition blocked light-induced spheroid enlargement. This is indicative of border cells generating outward-directed traction forces on the surrounding extracellular matrix (ECM), driving spheroid expansion.
Finding 3: A Remote-Control Light Switch (Optogenetics)
The authors of this study worked out a way to use a remote-controlled light switch to influence the activity of the station manager (GEF-H1). Now, when the authors shine blue light on the station manager, she speeds up the escalators! This results in a sudden burst in foot traffic toward the exits, and more people start escaping the train station. Experimental data show that long-term optogenetic stimulation led to the escape of individual melanoma cells from the 3D-spheroids into the surrounding matrix—an in vitro model of invasive behaviour. This presents direct evidence for the specific role increased contraction dynamics play in promoting invasiveness.
The big picture.
This entire process illustrates that despite the lack of external commands dictating people to leave, the internal control room (GEF-H1) can still generate force and the required direction for people to escape. The preprinted study highlighted here beautifully elucidates how cancer cells might break away from tumors and start invading other tissues—even in absence of external signals. The study establishes that GEF-H1-induced Rho activity, and subsequently cell contraction, could be a key step during tumor progression. Additionally, findings from this study offer insights into how dysregulated Rho signaling could contribute to cancer metastasis and underscore GEFs as potential therapeutic targets.
What do I like most about the preprint
I really like that this study associates signal dynamics with tumor invasive behaviors. Rather than monitoring only static alterations, this study stresses the importance of dynamic, pulsatile contraction and its effect on invasion. This spatiotemporal aspect of signaling in the framework of cell mechanics is often ignored and is essential to our understanding of tumor biology. Furthermore, a mechanistic explanation of asymmetric force generation at the spheroid boundary is a key conceptual advance. It depicts how tumor cells could “pull” themselves away from the tumor bulk during invasion.
Significance of the study
In-depth knowledge of how mechanical forces navigate tumor invasion is fundamental to developing new therapeutic strategies. The authors of this study have established a mouse melanoma model allowing the precise light-controlled modulation of Rho activity and myosin-based contraction dynamics.
- This study successfully establishes how enhanced and pulsatile cell contraction dynamics promote invasive tumor behavior in both 2D cultures and 3D multicellular tumor spheroids (MCTS). Increased contractility with precise optogenetics activation of Lbc-type GEF, GEF-H1, results in amplified focal adhesion turnover, force asymmetry at the spheroid border, and eventual detachment and escape of individual tumor cells.
- This observation reveals the existence of spatio-temporal regulation by GEF-H1 on contraction dynamics, promoting matrix-degradation–independent amoeboid-like invasion.
- Further, it indicates that targeting Rho signaling pathways could be a promising strategy to limit cancer cell dissemination.
doi: https://doi.org/10.1242/prelights.40283
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