Photothermal conversion in the second near-infrared (NIR-II) window allows deeper penetration and higher exposure to lasers, but examples of NIR-II photothermal agents are mainly formulated by inorganic compounds. In view of the underlying influence of inorganic materials, a novel NIR-II photothermal nanoagent based on a narrow band gap D-A conjugated polymer (TBDOPV-DT) with 2,2-bithiophene as the donor and thiophene-fused benzodifurandione-based oligo( p-phenylenevinylene) as the acceptor has been developed. More importantly, TBDOPV-DT nanoparticles (TBDOPV-DT NPs) are demonstrated to combine excellent photoacoustic imaging (PAI) and photothermal therapy (PTT) ability. TBDOPV-DT NPs exhibit dramatic photostability and heating reproducibility with a photothermal conversion efficiency of 50%. Especially, the NPs possess a remarkable PTT effect toward cancer cells in vitro and can eliminate tumor cells completely in vivo under 1064 nm laser irradiation, while no appreciable side effects have been observed. This study achieves PAI-guided cancer therapy and sheds light on the future of using organic polymer NPs for the NIR-II PTT of cancer.