Skip to main content
2024 BMES Annual Meeting Main banner
Icon Legend
This session is not in your schedule.
This session is in your schedule. Click again to remove it.
Back

Cancer Technologies

Cancer Technologies - Poster Session E

Poster T3 - The regulation of cancer cell bioenergetics by its migration state

Saturday, October 26, 2024
10:00 AM – 11:00 AM EST
Location: Exhibit Hall E, F & G

Introduction:  

Introduction: Breast cancer cell metabolism is heterogeneous and dynamically adapts throughout the metastatic process, complicating the determination of the role of metabolic changes in cancer progression [1]. It is known that cell morphology affects metabolic activity in cancer cells, specifically the ATP:ADP ratio, playing a crucial part in regulating metabolic pathways of cancer cells [2], and intracellular ATP:ADP ratio has been positively correlated with migration potential [3]. Additionally, an altered metabolic profile has been identified as a driving force of mechanical regulation of cytoskeletal activity and traction force generation during migration [2]. However, it is still unclear how particular metabolic pathways contribute to regulating cytoskeletal activity for force generation and energy utilization during migration [4]. Here, micropatterning is used to allow or prevent migration to investigate the effects of migration on single cell metabolic reprogramming and activity. This information can aid in directing the development of therapies that target cellular metabolism to prevent metastasis.

Materials and

Methods:  Materials and

Methods: MDA-MB-231 breast cancer cells were seeded onto 2D linear polyacrylamide micropatterned slides, with a teardrop patterned and nonpatterned area. The teardrop pattern confined and prevented cells from migrating, while controlling cell morphology to resemble that of migrating cells. Confocal microscopy was utilized to investigate single cells at 12 hours post seeding. The PercevalHR fluorescent sensor was used to measure ATP:ADP ratio, and pHRed fluorescent protein was utilized for pH correction. The genetically encoded fluorescent biosensor Peredox-mCherry was used to determine cytosolic NADH:NAD+ redox state. Fluorescent dyes TMRM and 2-NBDG were used to measure mitochondrial membrane potential and glucose uptake, respectively.

Results, Conclusions, and Discussions: 

Results: Nonmigrating MDA-MB-231 cells had an increased ATP:ADP ratio measured by the PercevalHR probe after pH correction and mitochondrial membrane potential (Fig. 1A and 1B), consistent with our previous work for cells confined on square micropatterns. In addition, nonmigrating cells had an increase in glucose uptake (Fig. 1C) compared to migrating cells. By contrast, no significant difference was observed for the Peredox-mCherry biosensor, a metric of cellular NADH:NAD+ ratio (Fig. 1D).

Discussions: Nonmigrating confined cells show an increase in pH corrected ATP:ADP ratio and mitochondrial membrane potential as described previously. [2] Here, increased glucose uptake, in nonmigrating cells, is a potential rationalization for the results reported by the Peredox-mCherry probe. This data indicates that in nonmigrating cells both metabolic pathways for ATP production, oxidative phosphorylation and glycolysis, were increased compared to migrating cells without a reprogramming or shift of the metabolic pathways.

Conclusions: Confinement and prevention of migration leads to an increased mitochondrial membrane potential and glucose uptake, maintaining the cytosolic NADH:NAD+ ratio in single breast cancer cells. Furthermore, nonmigrating cells exhibit a higher ATP:ADP ratio, a potential indicator of metastatic and migration energy. Further understanding of the link between single cell metabolic dynamics and migration is essential for developing metastasis prevention therapeutics.

Acknowledgements (Optional):  

References:
1. Bergers, G., Fendt, SM. The metabolism of cancer cells during metastasis. Nat Rev Cancer 21, 162–180 (2021). https://doi.org/10.1038/s41568-020-00320-2
2. Wu Y, Zanotelli MR, Zhang J, Reinhart-King CA. Matrix-driven changes in metabolism support cytoskeletal activity to promote cell migration. Biophys J. 2021 May 4;120(9):1705-1717. doi: 10.1016/j.bpj.2021.02.044. Epub 2021 Mar 9. PMID: 33705759; PMCID: PMC8204337.
3. Zanotelli MR, Goldblatt ZE, Miller JP, Bordeleau F, Li J, VanderBurgh JA, Lampi MC, King MR, Reinhart-King CA. Regulation of ATP utilization during metastatic cell migration by collagen architecture. Mol Biol Cell. 2018 Jan 1;29(1):1-9. doi: 10.1091/mbc.E17-01-0041. Epub 2017 Nov 8. PMID: 29118073; PMCID: PMC5746062.
4. Zanotelli MR, Zhang J, Reinhart-King CA. Mechanoresponsive metabolism in cancer cell migration and metastasis. Cell Metab. 2021 Jul 6;33(7):1307-1321. doi: 10.1016/j.cmet.2021.04.002. Epub 2021 Apr 28. PMID: 33915111; PMCID: PMC9015673.