Enzyme-Instructed Peptide Assembly Favored by Preorganization for Cancer Cell Membrane Engineering.

Area of Science:

• Biomaterials Science
• Chemical Biology
• Cancer Research

Background:

• Cancer cell membrane engineering is crucial for developing advanced cell-based therapeutics and manipulating cell-cell interactions.
• Current methods for modifying cancer cell membranes require further innovation for targeted and efficient therapeutic applications.

Purpose of the Study:

• To develop an *in situ* method for selective cancer cell membrane modification using enzyme-instructed peptide self-assembly (EISA).
• To investigate how specific phosphorylation patterns on peptides influence their self-assembly behavior and targeting of the epidermal growth factor receptor (EGFR).
• To explore the potential of EISA for creating membrane-bound peptide assemblies for cancer immunotherapy.

Main Methods:

• Utilized three phosphopeptides (pY1, pY2, pY3) targeting membrane-bound EGFR, differing in a single phosphorylated tyrosine.
• Employed an EISA strategy to induce self-assembly of peptides on cancer cell membranes.
• Investigated peptide preorganization, self-assembly kinetics, and spatial distribution *in cellulo*.
• Assessed peptide binding affinity to EGFR after dephosphorylation by alkaline phosphatase (ALP).
• Applied peptide-protein and peptide-peptide co-assembly to immobilize antigens (ovalbumin and dinitrophenyl hapten) on cancer cell membranes.

Main Results:

• Site-specific phosphorylation patterns distinctly controlled peptide preorganization, self-assembling kinetics, and spatial distribution.
• pY1 demonstrated superior preorganization, faster dephosphorylation by ALP, and higher binding affinity for EGFR post-dephosphorylation.
• Stable peptide assemblies were successfully constructed on cancer cell membranes using pY1 with ALP and EGFR.
• Antigens were effectively localized on cancer cell membranes via co-assembly strategies.

Conclusions:

• EISA provides a versatile technique for *in situ* construction of membrane-bound peptide assemblies on cancer cells.
• This method offers a promising strategy for artificially enriching cancer cell membrane components for potential cancer immunotherapy.
• Targeted engineering of cancer cell membranes can significantly advance the development of novel cancer therapeutics.