Background: Hepatocellular carcinoma(HCC) is one of the most highly malignant tumors. Liver tumor-initiating cells(LTICs) have been considered to contribute to HCC progression and metastasis. ATP-citrate lyase(ACLY), ...Background: Hepatocellular carcinoma(HCC) is one of the most highly malignant tumors. Liver tumor-initiating cells(LTICs) have been considered to contribute to HCC progression and metastasis. ATP-citrate lyase(ACLY), as a key enzyme for de novo lipogenesis, has been reported to be upregulated in various tumors. However, its expression and role in HCC and LTICs remain unknown. Methods: The expressions of ACLY in HCC tissues were detected by quantitative real-time PCR(q RT-PCR), Western blotting and immunohistochemistry. Kaplan-Meier curves and Chi-square test were used to determine the clinical significance of ACLY expression in HCC patients. A series of assays were performed to determine the function of ACLY on stemness, migration and invasion of HCC cells. Luciferase reporter assay, Western blotting and immunoprecipitation were used to study the regulation of the Wnt/β-catenin signaling by ACLY. Rescue experiments were performed to investigate whether β-catenin was the mediator of ACLY-regulated stemness and migration in HCC cells. Results: ACLY was highly expressed in HCC tissues and LTICs. Overexpression of ACLY was significantly correlated with poor prognosis, progression and metastasis of HCC patients. Knockdown of ACLY remarkably suppressed stemness properties, migration and invasion in HCC cells. Mechanistically, ACLY could regulate the canonical Wnt pathway by affecting the stability of β-catenin, and Lys49 acetylation of β-catenin might mediate ACLY-regulated β-catenin level in HCC cells. Conclusions: ACLY is a potent regulator of Wnt/β-catenin signaling in modulating LTICs stemness and metastasis in HCC. ACLY may serve as a new target for the diagnosis and treatment of HCC.展开更多
Adenosine triphosphate(ATP)-citrate lyase(ACLY)converts cytosolic citrate from the tricarboxylic acid cycle(TCA cycle)into acetyl coenzyme A(acetyl-CoA),which is a building block for cholesterol and fatty acid synthes...Adenosine triphosphate(ATP)-citrate lyase(ACLY)converts cytosolic citrate from the tricarboxylic acid cycle(TCA cycle)into acetyl coenzyme A(acetyl-CoA),which is a building block for cholesterol and fatty acid synthesis.Abnormally high expression of ACLY is associated with multiple metabolic diseases including dyslipidemia,atherosclerosis and non-alcoholic steatohepatitis(NASH),making ACLY an appealing target.In previous work,326Eb was discovered,featuring an alkenyl dicarboxylic acid scaffold as a novel ACLY inhibitor.326E can be potentially used to treat hypercholesterolemia and is currently undergoing phase II clinical trials.In this study,a series of diacids containing conjugated diene were developed based on impurities in manufacturing process of 326E in good manufacturing practice of medical products(GMP)condition,which represented a unique structural type different from known ACLY inhibitors.Among synthesized all eight possible isomers,compounds 43ZZ and 52EE exhibited significant inhibitory effect on de novo lipogenesis and gluconeogenesis in vitro and 43ZZ showed favorable pharmacokinetics.Furthermore,oral administration of 43ZZ and 52EE demonstrated a marked reduction of hepatic lipogenesis,along with lowered plasma levels of triglyceride and cholesterol,thereby confirming that these diene diacids as novel ACLY inhibitors have therapeutic potential for hyperlipidemia.展开更多
Hepatic cholesterol accumulation is an important contributor to hypercholesterolemia,which results in atherosclerosis and cardiovascular disease(CVD).ATP-citrate lyase(ACLY)is a key lipogenic enzyme that converts cyto...Hepatic cholesterol accumulation is an important contributor to hypercholesterolemia,which results in atherosclerosis and cardiovascular disease(CVD).ATP-citrate lyase(ACLY)is a key lipogenic enzyme that converts cytosolic citrate derived from tricarboxylic acid cycle(TCA cycle)to acetyl-CoA in the cytoplasm.Therefore,ACLY represents a link between mitochondria oxidative phosphorylation and cytosolic de novo lipogenesis.In this study,we developed the small molecule 326E with an enedioic acid structural moiety as a novel ACLY inhibitor,and its CoA-conjugated form 326E-CoA inhibited ACLY activity with an IC_(50)=5.31±1.2μmol/L in vitro.326E treatment reduced de novo lipogenesis,and increased cholesterol efflux in vitro and in vivo.326E was rapidly absorbed after oral administration,exhibited a higher blood exposure than that of the approved ACLY inhibitor bempedoic acid(BA)used for hypercholesterolemia.Chronic 326E treatment in hamsters and rhesus monkeys resulted in remarkable improvement of hyperlipidemia.Once daily oral administration of 326E for 24 weeks prevented the occurrence of atherosclerosis in ApoE^(-/-)mice to a greater extent than that of BA treatment.Taken together,our data suggest that inhibition of ACLY by 326E represents a promising strategy for the treatment of hypercholesterolemia.展开更多
Twelve new cis-clerodane diterpenoids, designated as crispinoids A—L (1—12), together with three known analogues (13—15), were isolated from Tinospora crispa. Their structures were fully assigned by comprehensive s...Twelve new cis-clerodane diterpenoids, designated as crispinoids A—L (1—12), together with three known analogues (13—15), were isolated from Tinospora crispa. Their structures were fully assigned by comprehensive spectroscopic techniques, single-crystal X-ray diffraction experiments, and electronic circular dichroism (ECD) analyses. The isolated clerodanes displayed diverse heterocyclic frameworks including 6/6/6-, 6/5/6/6-, 6/6/5-, 6/6-, and 6/5/6-fused ring systems. Some of the isolates showed ATP-citrate lyase (ACLY) and nuclear factor kappa B (NF-κB) pathway inhibition. The NF-κB inhibitors further suppressed the lipopolysaccharide (LPS)-induced inflammatory responses in RAW 264.7 cells.展开更多
基金supported by grants from the National Natu-ral Science Foundation of China (81972779)Ministry of Education (MOE) Key Laboratory on signaling Regulation and Targeting Therapy of Liver Cancer,and Shanghai Key Laboratory of Hepato-biliary Tumor Biology,Chinese National Key Project (2018ZX10723204-006-003)。
文摘Background: Hepatocellular carcinoma(HCC) is one of the most highly malignant tumors. Liver tumor-initiating cells(LTICs) have been considered to contribute to HCC progression and metastasis. ATP-citrate lyase(ACLY), as a key enzyme for de novo lipogenesis, has been reported to be upregulated in various tumors. However, its expression and role in HCC and LTICs remain unknown. Methods: The expressions of ACLY in HCC tissues were detected by quantitative real-time PCR(q RT-PCR), Western blotting and immunohistochemistry. Kaplan-Meier curves and Chi-square test were used to determine the clinical significance of ACLY expression in HCC patients. A series of assays were performed to determine the function of ACLY on stemness, migration and invasion of HCC cells. Luciferase reporter assay, Western blotting and immunoprecipitation were used to study the regulation of the Wnt/β-catenin signaling by ACLY. Rescue experiments were performed to investigate whether β-catenin was the mediator of ACLY-regulated stemness and migration in HCC cells. Results: ACLY was highly expressed in HCC tissues and LTICs. Overexpression of ACLY was significantly correlated with poor prognosis, progression and metastasis of HCC patients. Knockdown of ACLY remarkably suppressed stemness properties, migration and invasion in HCC cells. Mechanistically, ACLY could regulate the canonical Wnt pathway by affecting the stability of β-catenin, and Lys49 acetylation of β-catenin might mediate ACLY-regulated β-catenin level in HCC cells. Conclusions: ACLY is a potent regulator of Wnt/β-catenin signaling in modulating LTICs stemness and metastasis in HCC. ACLY may serve as a new target for the diagnosis and treatment of HCC.
文摘Adenosine triphosphate(ATP)-citrate lyase(ACLY)converts cytosolic citrate from the tricarboxylic acid cycle(TCA cycle)into acetyl coenzyme A(acetyl-CoA),which is a building block for cholesterol and fatty acid synthesis.Abnormally high expression of ACLY is associated with multiple metabolic diseases including dyslipidemia,atherosclerosis and non-alcoholic steatohepatitis(NASH),making ACLY an appealing target.In previous work,326Eb was discovered,featuring an alkenyl dicarboxylic acid scaffold as a novel ACLY inhibitor.326E can be potentially used to treat hypercholesterolemia and is currently undergoing phase II clinical trials.In this study,a series of diacids containing conjugated diene were developed based on impurities in manufacturing process of 326E in good manufacturing practice of medical products(GMP)condition,which represented a unique structural type different from known ACLY inhibitors.Among synthesized all eight possible isomers,compounds 43ZZ and 52EE exhibited significant inhibitory effect on de novo lipogenesis and gluconeogenesis in vitro and 43ZZ showed favorable pharmacokinetics.Furthermore,oral administration of 43ZZ and 52EE demonstrated a marked reduction of hepatic lipogenesis,along with lowered plasma levels of triglyceride and cholesterol,thereby confirming that these diene diacids as novel ACLY inhibitors have therapeutic potential for hyperlipidemia.
基金supported by grants from the National Natural Science Foundation of China(92057116 and 82170872)the Medical Guidance Project of Shanghai Science and Technology Commission(20S11903400,China)+3 种基金the“Personalized Medicines-Molecular Signature-based Drug Discovery and Development”Strategic Priority Research Program of the Chinese Academy of Sciences(XDA12040328,China)the“State Key Laboratory of Drug Research”Shanghai Institute of Materia Medica,Chinese Academy of Sciences(SIMM2105KF-02,China)Natural Science Foundation of Shanghai’s 2021“Science and Technology Innovation Action Plan”(21ZR1475300,China)the Lingang Laboratory(LG-QS-202205-01,China)。
文摘Hepatic cholesterol accumulation is an important contributor to hypercholesterolemia,which results in atherosclerosis and cardiovascular disease(CVD).ATP-citrate lyase(ACLY)is a key lipogenic enzyme that converts cytosolic citrate derived from tricarboxylic acid cycle(TCA cycle)to acetyl-CoA in the cytoplasm.Therefore,ACLY represents a link between mitochondria oxidative phosphorylation and cytosolic de novo lipogenesis.In this study,we developed the small molecule 326E with an enedioic acid structural moiety as a novel ACLY inhibitor,and its CoA-conjugated form 326E-CoA inhibited ACLY activity with an IC_(50)=5.31±1.2μmol/L in vitro.326E treatment reduced de novo lipogenesis,and increased cholesterol efflux in vitro and in vivo.326E was rapidly absorbed after oral administration,exhibited a higher blood exposure than that of the approved ACLY inhibitor bempedoic acid(BA)used for hypercholesterolemia.Chronic 326E treatment in hamsters and rhesus monkeys resulted in remarkable improvement of hyperlipidemia.Once daily oral administration of 326E for 24 weeks prevented the occurrence of atherosclerosis in ApoE^(-/-)mice to a greater extent than that of BA treatment.Taken together,our data suggest that inhibition of ACLY by 326E represents a promising strategy for the treatment of hypercholesterolemia.
基金supported by the National Natural Science Foundation of China(No.22277074).
文摘Twelve new cis-clerodane diterpenoids, designated as crispinoids A—L (1—12), together with three known analogues (13—15), were isolated from Tinospora crispa. Their structures were fully assigned by comprehensive spectroscopic techniques, single-crystal X-ray diffraction experiments, and electronic circular dichroism (ECD) analyses. The isolated clerodanes displayed diverse heterocyclic frameworks including 6/6/6-, 6/5/6/6-, 6/6/5-, 6/6-, and 6/5/6-fused ring systems. Some of the isolates showed ATP-citrate lyase (ACLY) and nuclear factor kappa B (NF-κB) pathway inhibition. The NF-κB inhibitors further suppressed the lipopolysaccharide (LPS)-induced inflammatory responses in RAW 264.7 cells.
