Tumor-Treating Fields Alter Nanomechanical Properties of Pancreatic Ductal Adenocarcinoma Cells Co-Cultured with Extracellular Matrix

Our plain-English summary of Tumor-Treating Fields Alter Nanomechanical Properties of Pancreatic Ductal Adenocarcinoma Cells Co-Cultured with Extracellular Matrix, by T. Kulkarni, S. Banik, D. Mukhopadhyay, et al (Journal of Functional Biomaterials, 2025).

The study

• Authors: T. Kulkarni, S. Banik, D. Mukhopadhyay, H. Babiker, S. Bhattacharya
• Journal: Journal of Functional Biomaterials
• Published: 2025;16(5)
• Cited by: 2 articles, as counted by NIH iCite
• Read it: PubMed · Publisher · Free full text

Abstract

Tumor-Treating Fields (TTFields), a novel therapeutic avenue, is approved for therapy in Glioblastoma multiforme, malignant pleural mesothelioma, and metastatic non-small cell lung cancer (NSCLC). In pancreatic ductal adenocarcinoma (PDAC), several clinical trials are underway to improve outcomes, yet a significant knowledge gap prevails involving the cell-extracellular matrix (ECM) crosstalk. Herein, we hypothesized that treatment with TTFields influence this crosstalk, which is reflected by the dynamic alteration in nanomechanical properties (NMPs) of cells and the ECM in a co-culture system. We employed an ECM gel comprising collagen, fibronectin, and laminin mixed in 100:1:1 stoichiometry to co-culture of Panc1 and AsPC1 individually. This ECM mixture mimics the in vivo tumor microenvironment closely when compared to the individual ECM components studied before. A comprehensive frequency-dependent study revealed the optimal TTFields frequency to be 150 kHz. We also observed that irrespective of the ECM's presence, TTFields increase cell membrane stiffness and decrease deformation several-folds in both Panc1 and AsPC1 cells at both 48 h and 72 h. Although adhesion for AsPC1 decreased at 48 h, at 72 h it was observed to increase irrespective of ECM's presence. Moreover, it significantly alters the NMPs of ECM gels when co-cultured with PDAC cell lines. However, AsPC1 cells were observed to be more detrimental to these changes. Lastly, we attribute the stiffness changes in Panc1 cells to the membrane F-actin reorganization in the presence of TTFields. This study paves a path to study complex PDAC TME as well as the effect of various chemotherapeutic agents on such TME with TTFields in the future.

In plain English

This laboratory study did not examine asbestos or mesothelioma. Malignant pleural mesothelioma appeared only in the abstract and introduction, as an approved use of Tumor-Treating Fields (TTFields), which are alternating electric fields, and as the subject of an ongoing trial. The authors grew pancreatic ductal adenocarcinoma cell lines Panc1 and AsPC1 alone or on a 100:1:1 collagen, fibronectin and laminin gel. They applied 150 kHz TTFields and measured stiffness, deformation and adhesion by atomic force microscopy at 48 and 72 hours. Panc1 stiffness averaged 3.7 and 4.1 kPa untreated, versus 21 and 24.4 kPa treated, at 48 and 72 hours. AsPC1 stiffness went from 5.3 and 5.1 kPa untreated to 11.5 and 16.6 kPa treated. Deformation fell in both cell lines. In co-cultures, TTFields lowered adhesion at 48 hours and raised it at 72 hours. The gel grown with either cell line stiffened under TTFields; without cells, the three-protein gel had decreased in stiffness at 150 kHz. Treated Panc1 cells showed less F-actin staining; the authors attributed Panc1 stiffness changes to actin reorganization. They acknowledged their matrix model was simplistic. Novocure partly funded the study.

We wrote this plain-English summary from the full article with the help of AI, and a second AI model checked each sentence against the article. It is not the authors' wording; the abstract and the article are their own account.

What the study did and found

• Design: Laboratory (in vitro) cell-culture experiment using atomic force microscopy and confocal microscopy
• Who was studied: Two pancreatic ductal adenocarcinoma cell lines, Panc1 (primary) and AsPC1 (metastatic), grown alone or on a collagen, fibronectin and laminin gel (100:1:1); at least 12 cells probed per condition
• Exposure: Tumor-Treating Fields (alternating electric fields) at 150 kHz for 48 h and 72 h; frequencies from 50 to 300 kHz were tested on matrix gels alone. No asbestos exposure was studied.
• Main results:
  • Panc1 cells alone: average membrane stiffness was 3.7 and 4.1 kPa untreated at 48 h and 72 h, and rose to 21 and 24.4 kPa with TTFields
  • Panc1 cells alone: deformation fell from 393.6 and 415.4 nm untreated to 120.9 and 77.7 nm with TTFields at 48 h and 72 h
  • AsPC1 cells alone: stiffness went from 5.3 and 5.1 kPa untreated to 11.5 and 16.6 kPa with TTFields at 48 h and 72 h
  • AsPC1 cells alone: adhesion at 48 h went from 692.5 pN untreated to 2768 pN with TTFields
  • Panc1 cells on the matrix gel: stiffness went from 5.4 kPa (48 h) and 3.6 kPa (72 h) untreated to 35.2 and 16.5 kPa with TTFields
  • AsPC1 cells on the matrix gel: stiffness went from 8.9 and 13.1 kPa untreated to 15 and 22 kPa with TTFields at 48 h and 72 h
  • Three-protein matrix gel without cells: stiffness decreased at 150 kHz
  • Matrix gel with Panc1 cells: gel stiffness went from 16.5 and 17.9 kPa untreated to 23.5 and 22.8 kPa with TTFields at 48 h and 72 h
  • Matrix gel with AsPC1 cells: gel stiffness went from 12 and 10 kPa untreated to 33.9 and 42.2 kPa with TTFields at 48 h and 72 h
  • In cell and matrix co-cultures, adhesion was lower with TTFields at 48 h and higher at 72 h
  • Panc1 cells: F-actin staining signal was significantly lower with TTFields at both 48 h and 72 h
• The authors' conclusion: The authors concluded that TTFields altered the nanomechanical properties of cells and matrix in co-culture. They attributed the stiffness changes in Panc1 cells, the only cells examined for actin, to F-actin reorganization.
• Limitations the authors state:
  • The authors stated that their matrix model is simplistic and far from a real tumor microenvironment
  • The authors stated that the cause of the matrix stiffness increase is not known
  • The authors stated that the mechanism behind the adhesion trends needs further work

Topics

Medical terms in this study, explained in our Lexicon: Adenocarcinoma, Pleural Mesothelioma, Non-Small Cell Lung Cancer, Clinical Trial, Collagen, Tumor Microenvironment.

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