Fmoc-FF/RGD peptide hydrogel rapidly induces cancer stemness, enables in situ NO monitoring, and direct drug evaluation in MCF-7 cells.

Understanding and targeting cancer stem cells (CSCs) is crucial for overcoming tumor progression, recurrence, and therapy resistance. However, conventional CSC models, which often rely on sphere formation, are plagued by prolonged workflows (15-30 days), destructive characterization, and re-culture steps. These limitations hinder rapid, reproducible studies of CSC biology and efficient screening of CSC-targeting drugs. There is a critical need for integrated platforms that can streamline CSC induction, real-time pathway monitoring, and direct therapeutic evaluation within a single, efficient system.

Researchers engineered a functional peptide hydrogel interface by self-assembling Fmoc-diphenylalanine (Fmoc-FF) peptides. This 3D microenvironment was further enhanced by co-assembly with Fmoc-RGD peptides to activate integrin-mediated signaling. They applied this system to MCF-7 cells to induce stemness reprogramming within 3 days. To enable real-time monitoring, a manganese porphyrin was incorporated as a catalytic recognition element, allowing electrochemical detection of nitric oxide (NO) at the hydrogel-electrode interface. This NO-responsive platform was then used for direct evaluation of CSC-targeting drugs, eliminating the need for traditional sphere transfer and lysis-based assays.

The engineered peptide hydrogel successfully induced highly efficient stemness reprogramming of MCF-7 cells within a remarkably short period of 3 days. This rapid induction significantly reduced the overall experimental workflow to 6-8 days, a substantial improvement over conventional methods that typically require 15-30 days. The integrated manganese porphyrin element enabled real-time, in situ electrochemical detection of nitric oxide (NO), providing a functional readout of pathway activation during CSC induction. This novel design allowed for direct evaluation of CSC-targeting drugs within the same 3D system, bypassing the need for cumbersome sphere transfer and lysis-based assays. The integrated strategy demonstrated improved operational simplicity and enhanced functional readout capabilities.

The integrated strategy significantly shortens the experimental workflow to 6-8 days compared to conventional methods (15-30 days), while improving operational simplicity and functional readout capability.