Glucose oxidase-engineered cell membrane vesicles co-delivering CRISPR/Cas9 and croconaine dye for traceable synergistic tumor therapy

Mild photothermal therapy (PTT, <45 °C) minimizes collateral damage to healthy tissues but is frequently compromised by heat shock protein (HSP)-mediated thermal resistance. Herein, we develop a genetically engineered biomimetic nanoplatform, designated as pCas9@GM-Croc (CGMC), to achieve traceable synergistic starvation/gene/photothermal therapy. Glucose oxidase (GOx) is stably expressed on HEK293T cell membranes to fabricate vesicles (GM) that prevent premature enzyme leakage while ensuring robust lysosomal escape. These vesicles deliver clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) plasmids (pCas9) targeting HSP90α and integrate an acid-responsive croconaine dye (Croc) into their lipid bilayers. Mechanistically, GOx-mediated adenosine triphosphate (ATP) depletion acts in tandem with pCas9 genome editing to decrease HSP90α expression, profoundly impairing the thermal tolerance of tumor cells. Concurrently, GOx-catalyzed gluconic acid generation activates the fluorescence (FL) and photoacoustic (PA) signals of Croc, enabling real-time, non-invasive monitoring of therapeutic progression and tumor deoxygenation. Both in vitro and in vivo results demonstrate that CGMC exhibits high tumor accumulation and remarkable synergistic antitumor efficacy, achieving nearly complete tumor suppression under 808 nm laser irradiation with favorable biocompatibility. This work provides an imaging-traceable biomimetic strategy to overcome thermal endurance for enhanced tumor treatment.