In vivo membrane engineering traps Gd-based MRI contrast agents for detecting microhepatocellular carcinoma

Accurate detection of microhepatocellular carcinoma (HCC) remains a major clinical challenge owing to the limited specificity and sensitivity of current imaging modalities. Herein, we present a dual-injection magnetic resonance imaging (MRI) peptidic probe based on in vivo membrane engineering, achieving in situ signal amplification and molecularly precise imaging. The first injection of programmable nanoparticles coassembled from two peptide monomers, incorporating a GPC3-targeting ligand, a β sheet–forming motif, a dibenzocyclooctyne (DBCO) handle, and porphyrin IX (PpIX) for fluorescence tracking. Following systemic administration, the nanoparticles high-specifically bind to GPC3-overexpressing tumor membranes and transform into surface-anchored nanofibrils, exposing confined DBCO groups. A second injection of azide-modified Gd-DOTA enables rapid copper-free click conjugation on the nanofibrillar scaffold, yielding a nearly fourfold increase in longitudinal relaxivity. MRI demonstrated strong T 1 -weighted signal enhancement and high tumor-to-liver contrast in Hepa1-6 tumor-bearing mice. In vivo membrane engineering strategy establishes a generalizable platform for receptor-guided molecular imaging and early cancer detection.