Systemic Nanomechanical Single-Cell Profiling Reveals Mechanophenotype Transitions Under Therapeutic Perturbation

Area of Science:

• Biophysics
• Cancer Biology
• Cell Mechanics

Background:

• Mechanical remodeling of cancer cells influences their invasive behavior.
• The quantitative link between cell mechanics and therapeutic response is not well understood.

Purpose of the Study:

• To systematically characterize drug-induced changes in cancer cell mechanophenotypes.
• To establish a quantitative relationship between cell mechanical properties and therapeutic response.

Main Methods:

• Integrated nanomechanical profiling using atomic force microscopy (AFM) force mapping.
• High-resolution imaging and cytomorphometric analysis of fixed cells.
• Multivariate analysis and dimensionality reduction (PCA, PLS-DA) of biophysical parameters.

Main Results:

• Drug treatment caused significant cytoskeletal reorganization, increased cortical stiffness, and surface roughness.
• Eleven biophysical parameters were linked to invasive capacity, with nucleus modulus, cytoskeletal density, and cortical roughness being key.
• A distinct mechanophenotype transition was identified, characterized by increased stiffness and reduced protrusive activity.
• Reduced invasiveness correlated with increased cortical roughness and perinuclear cytoskeletal reorganization, forming a mechanical signature of phenotypic reprogramming.

Conclusions:

• Nanomechanical profiling provides a quantitative framework for assessing drug-induced phenotypic transitions in cancer cells.
• This approach can discriminate between invasive and noninvasive states at the single-cell level.
• Mechanical profiling offers a complementary method to molecular assays for evaluating cancer therapeutic response.