Targeting NAD + biosynthesis suppresses TGF-β1/Smads/RAB26 axis and potentiates cisplatin cytotoxicity in non-small cell lung cancer brain metastasis

Nicotinamide adenine dinucleotide (NAD⁺) plays an important role in tumor progression, but its role in non-small cell lung cancer with brain metastasis (NSCLC BM) remains unclear. Herein, we investigated NAD⁺ biosynthesis targeting as a new therapeutic strategy for NSCLC BM. Therapeutic activity of nicotinamide phosphoribosyl transferase (NAMPT) inhibitors was evaluated in mouse models of NSCLC BM and using various assays such as NAD⁺ quantitation, cell viability, and apoptosis assays. To explore impact on downstream signaling, RNA sequencing was used in NAMPT inhibitor-treated and control cells, followed by validation with genetic knockdown, western blot and qRT-PCR. Expression of NAMPT and downstream proteins in human NSCLC BM and its association with patient prognosis were examined. Finally, combination of NAMPT inhibitor and cisplatin was tested in vivo. Systemic treatment with NAMPT inhibitors demonstrated intracranial activity in an NSCLC BM model. NAMPT inhibitors decreased cellular NAD levels and suppressed proliferation and invasion, and induced apoptosis in NSCLC cells. Supplementation with NAD⁺ precursor NMN rescued these NAMPT inhibitor effects. Mechanistically, disruption of NAMPT-mediated NAD⁺ biosynthesis suppressed TGF-β1/Smads/RAB26 signaling, leading to inhibition of NSCLC cells. Expression of NAMPT/TGF-β1/Smads/RAB26 axis proteins was upregulated in NSCLC BM tissues and correlated with poor prognosis. Combining NAMPT inhibitors with cisplatin further extended the survival of NSCLC BM-bearing mice. Targeting NAD⁺ biosynthesis provides a new therapeutic strategy for NSCLC BM and can be effectively combined with cisplatin. Our studies identified the TGF-β1/Smads/RAB26 signaling downstream of NAMPT, which was targeted by NAMPT inhibition to mediate anti-cancer effects.