Dynamic Profiling and Screening of Cancer Starvation-Immunotherapy Regimens Via a Cross-Referencing Multispectral Upconversion and Tomographic Photoacoustic Imaging Nanoprobe.

Combining tumor-associated macrophage (TAM)-targeted immunotherapy with starvation therapy represents a promising strategy for enhancing antitumor efficacy. Energy metabolism and nitrosative stress represent two key metabolic pathways that can effectively guide the efficacy of the combination of TAM-targeted immunotherapy and starvation therapy. However, methods capable of simultaneously profiling of the dynamic interplay between energy metabolism and nitrosative stress during cancer starvation-immunotherapy (CSI) remains challenging. Here, we report a TAM-targeted dual-mode nanoprobe, NNDA, for real-time imaging of energy metabolism and M1-like TAM-mediated nitrosative stress during CSI. The nanoprobe was designed to comprise NAD(P)H-sensitive dye Glu-RB and NO-responsive dye CY-NO assembled on the surface of upconversion nanoparticles, showing excellent selectivity and sensitivity to energy metabolism-associated NAD(P)H and nitrosative stress-associated NO with two independent NIR photoacoustic (PA, 710/1064 nm) and upconversion luminescence (UCL, 660/800 nm) channels. In vitro and in vivo studies confirmed that NNDA accurately tracked metabolic reprogramming and TAM repolarization, revealing that CSI regimens sustain nitrosative stress under energy suppression. Importantly, the ratio R-PA1064/PA710 serves as an early prognostic indicator of treatment outcome, displaying strong correlation with tumor growth inhibition. This work provides a cross-referencing imaging tool for dynamically deciphering metabolic-immune crosstalk, facilitating the screening and optimization of synergistic anticancer therapies.