NSC 663284

Impaired autophagy is increasingly recognized as a key contributor to the pathogenesis of metabolic dysfunction-associated steatotic liver disease (MASLD). However, its underlying molecular mechanisms remain largely undefined. Emerging evidence implicates epigenetic regulators in modulating autophagic pathways in metabolic diseases. Therefore, this study aimed to elucidate the role of a histone methyltransferase, nuclear receptor binding SET domain protein 2 (NSD2), in regulating autophagy and its contribution to MASLD progression.

NSD2 expression levels were evaluated in liver tissues from patients with MASLD and mouse models. Functional studies were conducted using hepatocyte-specific Nsd2 knockout and overexpression mouse models, along with cleavage under targets and tagmentation analysis in hepatocyte cell lines. Additionally, the effects of pharmacological NSD2 inhibition using NSC663284 were evaluated in human liver organoids. Autophagy, hepatic steatosis, and related epigenetic changes were assessed through molecular and histological techniques.

NSD2 expression was markedly elevated in both patient livers and murine models, correlating positively with disease severity. Hepatic NSD2 deficiency alleviated diet-induced autophagy impairment and steatosis, while NSD2 overexpression exacerbated these pathologies. Mechanistically, NSD2 epigenetically suppressed TFEB transcription by promoting trimethylation of histone H4 at lysine 20, impairing autophagy. Pharmacological inhibition of NSD2 with NSC663284 similarly alleviated hepatic steatosis in human liver organoids.

NSD2 acts as a key epigenetic suppressor of TFEB-mediated autophagy in the liver, promoting lipid accumulation and MASLD progression. Targeting NSD2 represents a promising therapeutic strategy for MASLD.

Cell division cycle 25 (CDC25) phosphatase is a key cell cycle regulator whose overexpression is linked to tumor malignancy and poor prognosis. Its inhibition suppresses cancer cell growth and induces cell death, making it a promising therapeutic target. In this study, we aimed to develop novel CDC25 inhibitors with therapeutic potential. Using NSC663284 (compound 1) as the lead compound, we synthesized and screened derivatives based on cell-viability assays. Among five cancer types tested, lung cancer emerged as the most suitable model. Notably, compound 6 exhibited the most potent inhibitory activity against CDC25 phosphatase. A clinical data analysis of a Clinical Proteome Tumor Analysis Consortium (CPTAC) or The Cancer Genome Atlas Lung Adenocarcinoma (TCGA-LUAD) dataset revealed that high expression levels of CDC25A and CDC25C were associated with a poor prognosis and were positively correlated with the T status. Moreover, CDC25C expression was positively associated with lymph node metastasis. In vitro, compound 6 exhibited stronger cytotoxicity and pro-apoptotic effects than compound 1, and significantly suppressed LUAD cell colony formation, migration, and invasion. In vivo, both compounds reduced tumor growth in H1975 xenograft mice, with compound 6 inducing more extensive tumor cell death. Furthermore, compound 6 also inhibited LUAD patient-derived organoid (PDO) growth. Combination studies of compound 6 with chemotherapeutic drugs indicated that compound 6 exhibited synergistic effects with those drugs in inhibiting the growth of LUAD cells. In conclusion, compound 6 is a promising lead compound for CDC25 inhibition and warrants further investigation as a potential therapeutic agent for LUAD treatment.