Biophysical Benchmarks of Malignancy in Primary Breast Tumor Cells
Final Report Abstract
The importance of cancer associated fibroblasts (CAFs) and their functional role in tumor biology is multi‐faceted. CAFs influence the TME by promoting tumor initiation, growth, progression, invasion and importantly metastasis. CAF production and secretion of cytokines, growth factors and MMPs are essential for regulation of the TME. In our study, we addressed the intrinsic biophysical properties of primary LUB CAFs and NORM MES cells. In summary, all of our findings regarding gene / protein expression along with 2D and 3D biophysical properties, support that LUB CAFs mainly stem from breast mesenchymal / fibroblasts, which are activated in the TME. Interestingly, LUB CAFs sustain their activated status in cell culture experiments. Due to the increased gene expression of many of the 96 key CAF associated genes along with protein expression defining CAFs, these genes represent the activated state above normal fibroblast / mesenchymal cells. Biophysical comparisons demonstrated a slightly but significantly higher 2D and 3D cell velocity, 2D and 3D persistence and 2D / 3D invasion distance, but a lower collective traction force generation of the LUB CAFs versus NORM MES cells. This demonstrates that collective force generation is a poor indicator for predicting collective migration, which is further highlighted by comparison with an established breast carcinoma cell line that has undergone EMT (the triple‐negative MDA‐MB‐231 cell line) and that displays even higher 2‐D and 3‐D invasiveness compared to LUB CAFs but generates much lower traction forces. Our data further support the idea that activation of EMT‐associated pathways are responsible for the enhanced 2D and 3D migratory / invasive properties in CAFs. However, the patient groups spanning lymph node negative to lymph node positive as well as patients with or without distant metastasis were too limited in numbers in order to investigate how these altered biophysical properties might be involved in metastasis formation. Also, we could not investigate the biophysical properties of the tumor‐derived primary epithelial cells, but we now have found suitable culture conditions to make such experiments feasible in the future. With this in hand, it will be important to perform experiments to investigate how LUB CAFs influence tumor epithelial cells, and vice versa. These experiments could involve co‐culture conditions as well as patient‐derived tumor xenograft (PDTX) models in mice to assess further functions, like tumor size changes and invasion/ metastasis.
Publications
- Bayesian model selection for complex dynamic systems. Nat Comm. 9:1803, 2018
Mark C, Metzner C, Lautscham L, Strissel PL, Strick R, Fabry B
(See online at https://doi.org/10.1038/s41467-018-04241-5) - Pressure‐driven collective growth mechanism of planar cell colonies. J Phys D, 51:304004, 2018
Metzner C, Lange J, Krauss P, Wunderling N, Übelacker J, Martin F, Fabry B
(See online at https://doi.org/10.1088/1361-6463/aace4c) - Breast Cancer Cells Adapt Contractile Forces to Overcome Steric Hindrance. Biophys J. 116:1305‐1312, 2019
Cóndor M, Mark C, Gerum RC, Grummel NC, Bauer A, García‐Aznar JM, Fabry B
(See online at https://doi.org/10.1016/j.bpj.2019.02.029) - Plasticity of patient‐matched normal mammary epithelial cells is dependent on autologous adipose‐derived stem cells. Sci Rep. 9:10722, 2019
Kengelbach‐Weigand A, Tasbihi K, Strissel PL, Schmid R, Marques JM, Beier JP, Beckmann MW, Strick R, Horch RE, Boos AM
(See online at https://doi.org/10.1038/s41598-019-47224-2) - Collective forces of tumor spheroids in three‐dimensional biopolymer networks. Elife. 2020 Apr 30;9:e51912
Mark C, Grundy TJ, Strissel PL, Böhringer D, Grummel N, Gerum R, Steinwachs J, Hack CC, Beckmann MW, Eckstein M, Strick R, O'Neill GM, Fabry B
(See online at https://doi.org/10.7554/elife.51912) - pyTFM: A tool for traction force and monolayer stress microscopy. PLoS Comput Biol. 2021 Jun 21;17(6):e1008364
Bauer A, Prechová M, Fischer L, Thievessen I, Gregor M, Fabry B
(See online at https://doi.org/10.1371/journal.pcbi.1008364)
