Background:Melatonin,a natural hormone secreted by the pineal gland,has been reported to exhibit antitumor properties through diverse mechanisms of action.However,the oncostatic function of melatonin on esophageal squ...Background:Melatonin,a natural hormone secreted by the pineal gland,has been reported to exhibit antitumor properties through diverse mechanisms of action.However,the oncostatic function of melatonin on esophageal squamous cell carcinoma(ESCC) remains elusive.This study was conducted to investigate the potential effect and underlying molecular mechanism of melatonin as single anticancer agent against ESCC cells.Methods:ESCC cell lines treated with or without melatonin were used in this study.In vitro colony formation and 5-Ethynyl-2’-deoxyuridine(EdU) incorporation assays,and nude mice tumor xenograft model were used to confirm the proliferative capacities of ESCC cells.RNA-seq,qPCR,Western blotting,recombinant lentivirus-mediated target gene overexpression or knockdown,plasmids transfection and co-IP were applied to investigate the underlying molecular mechanism by which melatonin inhibited ESCC cell growth.IHC staining on ESCC tissue microarray and further survival analyses were performed to explore the relationship between target genes’ expression and prognosis of ESCC.Results:Melatonin treatment dose-dependently inhibited the proliferative ability and the expression of histone deacetylase 7(HDAC7),c-Myc and ubiquitin-specific peptidase 10(USP10) in ESCC cells(P<0.05).The expressions of HDAC7,c-Myc and USP10 in tumors were significantly higher than the paired normal tissues from 148 ESCC patients(P<0.001).Then,the Kaplan-Meier survival analysis suggested that ESCC patients with high HDAC7,c-Myc or USP10levels predicted worse overall survival(log-rank P<0.001).Co-IP and Western blotting further revealed that HDAC7physically deacetylated and activated β-catenin thus promoting downstream target c-Myc gene transcription.Notably,our mechanistic study validated that HDAC7/β-catenin/c-Myc could form the positive feedback loop to enhance ESCC cell growth,and USP10 could deubiquitinate and stabilize HDAC7 protein in the ESCC cells.Additionally,we verified that inhibition of the HDAC7/β-catenin/c-Myc axis and USP10/HDAC7 pathway mediated the anti-proliferative action of melatonin on ESCC cells.Conclusions:Our findings elucidate that melatonin mitigates the HDAC7/β-catenin/c-Myc positive feedback loop and inhibits the USP10-maintained HDAC7 protein stability thus suppressing ESCC cell growth,and provides the reference for identifying biomarkers and therapeutic targets for ESCC.展开更多
HDAC7,a member of class IIa HDACs,plays a pivotal regulatory role in tumor,immune,fibrosis,and angiogenesis,rendering it a potential therapeutic target.Nevertheless,due to the high similarity in the enzyme active site...HDAC7,a member of class IIa HDACs,plays a pivotal regulatory role in tumor,immune,fibrosis,and angiogenesis,rendering it a potential therapeutic target.Nevertheless,due to the high similarity in the enzyme active sites of class IIa HDACs,inhibitors encounter challenges in discerning differences among them.Furthermore,the substitution of key residue in the active pocket of class IIa HDACs renders them pseudo-enzymes,leading to a limited impact of enzymatic inhibitors on their function.In this study,proteolysis targeting chimera(PROTAC)technology was employed to develop HDAC7 drugs.We developed an exceedingly selective HDAC7 PROTAC degrader B14 which showcased superior inhibitory effects on cell proliferation compared to TMP269 in various diffuse large B cell lymphoma(DLBCL)and acute myeloid leukemia(AML)cells.Subsequent investigations unveiled that B14 disrupts BCL6 forming a transcriptional inhibition complex by degrading HDAC7,thereby exerting proliferative inhibition in DLBCL.Our study broadened the understanding of the non-enzymatic functions of HDAC7 and underscored the importance of HDAC7 in the treatment of hematologic malignancies,particularly in DLBCL and AML.展开更多
Postnatal mammalian cardiomyocytes(CMs)rapidly lose proliferative capacity and exit the cell cycle to undergo further differentiation and maturation.Cell cycle activation has been a major strategy to stimulate postnat...Postnatal mammalian cardiomyocytes(CMs)rapidly lose proliferative capacity and exit the cell cycle to undergo further differentiation and maturation.Cell cycle activation has been a major strategy to stimulate postnatal CM proliferation,albeit achieving modest effects.One impediment is that postnatal CMs may need to undergo dedifferentiation before proliferation,if not simultaneously.Here,we report that overexpression of Hdac7 in neonatal mouse CMs results in significant CM dedifferentiation and proliferation.Mechanistically,we showthat histone deacetylase7(HDAC7)-mediatedCM proliferation is contingent on dedifferentiation,which is accomplished by suppressing myocyte enhancefactor2(MEF2).Hdac7overexpression in CM shifts the chromatin state from binding with MEF2,which favors the transcriptional program toward differentiation,to binding with AP-1,which favors the transcriptional program toward proliferation.Furthermore,we found that HDAC7 interacts with minichromosome maintenance complex components to initiate cell cycleprogression.Ourfindings revealthat HDAC7 promotes CM proliferation byits dual action on CM dedifferentiation and proliferation,uncovering a potential new strategy for heart regeneration/repair.展开更多
目的探讨白皮杉醇对脓毒症肺损伤小鼠代谢重编程的影响及可能机制。方法采用随机数字表法将32只雄性C57/B6小鼠随机分为4组,即对照组、模型组、低剂量治疗组和高剂量治疗组,每组各8只。对照组不做任何处理,其他3组使用脂多糖(lipopolysa...目的探讨白皮杉醇对脓毒症肺损伤小鼠代谢重编程的影响及可能机制。方法采用随机数字表法将32只雄性C57/B6小鼠随机分为4组,即对照组、模型组、低剂量治疗组和高剂量治疗组,每组各8只。对照组不做任何处理,其他3组使用脂多糖(lipopolysaccharide,LPS,10mg/kg)单次腹腔注射的方式构建脓毒症肺损伤小鼠模型,其中低剂量治疗组和高剂量治疗组小鼠于LPS注射前7天分别给予25mg/(kg·d)和100mg/(kg·d)的白皮杉醇灌胃,对照组和模型组给予等量0.9%氯化钠溶液灌胃。LPS腹腔注射12h后,留取小鼠支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)和肺组织,通过形态学和分子生物学实验评估各组小鼠肺损伤状况。结果与对照组比较,模型组小鼠肺组织病理损伤较对照组更明显,主要表现为肺水肿和炎性细胞浸润,肺损伤评分更高(8.25±0.45分vs 1.31±0.23分),BALF中乳酸含量(0.65±0.12mmol/L vs 0.17±0.04mmol/L)和乳酸盐脱氢酶活性(8934.12±145.25U/L vs 4782.53±111.04U/L)增加,肺组织中IL-1β、IL-6和TNF-α的mRNA水平明显升高,肺组织中糖酵解标志蛋白(GLUT1、HK2、PDK2、PKM2)和组蛋白去乙酰化酶7(histone deacetylase 7,HDAC7)的蛋白表达明显上调,差异均有统计学意义(P<0.05);与模型组比较,低剂量治疗组和高剂量治疗组小鼠肺病理损伤明显减轻,肺损伤评分更低(5.69±0.92分、2.04±0.31分vs 8.25±0.45分),BALF中乳酸含量(0.42±0.03mmol/L、0.25±0.02mmol/L vs 0.65±0.12mmol/L)和乳酸盐脱氢酶活性(6822.24±172.02U/L、5872.02±93.07U/L vs 8934.12±145.25U/L)降低,肺组织中IL-1β、IL-6和TNF-α的mRNA水平明显降低,肺组织中GLUT1、HK2、PDK2、PKM2和HDAC7的蛋白表达均明显下降,差异均有统计学意义(P<0.05)。结论白皮杉醇可显著减轻脓毒症小鼠肺病理损伤和炎性反应,其机制可能与HDAC7介导的代谢重编程抑制有关。展开更多
Aim:Multiple myeloma(MM)is a hematological malignancy of antibody-producing mature B cells or plasma cells.The proteasome inhibitor,bortezomib,was the first-in-class compound to be FDA approved for MM and is frequentl...Aim:Multiple myeloma(MM)is a hematological malignancy of antibody-producing mature B cells or plasma cells.The proteasome inhibitor,bortezomib,was the first-in-class compound to be FDA approved for MM and is frequently utilized in induction therapy.However,bortezomib refractory disease is a major clinical concern,and the efficacy of the pan-histone deacetylase inhibitor(HDACi),panobinostat,in bortezomib refractory disease indicates that HDAC targeting is a viable strategy.Here,we utilized isogenic bortezomib resistant models to profile HDAC expression and define baseline and HDACi-induced expression patterns of individual HDAC family members in sensitive vs.resistant cells to better understanding the potential for targeting these enzymes.Methods:Gene expression of HDAC family members in two sets of isogenic bortezomib sensitive or resistant myeloma cell lines was examined.These cell lines were subsequently treated with HDAC inhibitors:panobinostat or vorinostat,and HDAC expression was evaluated.CRISPR/Cas9 knockdown and pharmacological inhibition of specific HDAC family members were conducted.Results:Interestingly,HDAC6 and HDAC7 were significantly upregulated and downregulated,respectively,in bortezomib-resistant cells.Panobinostat was effective at inducing cell death in these lines and modulated HDAC expression in cell lines and patient samples.Knockdown of HDAC7 inhibited cell growth while pharmacologically inhibiting HDAC6 augmented cell death by panobinostat.Conclusion:Our data revealed heterogeneous expression of individual HDACs in bortezomib sensitive vs.resistant isogenic cell lines and patient samples treated with panobinostat.Cumulatively our findings highlight distinct roles for HDAC6 and HDAC7 in regulating cell death in the context of bortezomib resistance.展开更多
基金supported by the National Natural Science Foundation of China (82103508, 81871866, 82173252, 81672996)the Natural Science Foundation of Shaanxi Province (2022JQ?862)。
文摘Background:Melatonin,a natural hormone secreted by the pineal gland,has been reported to exhibit antitumor properties through diverse mechanisms of action.However,the oncostatic function of melatonin on esophageal squamous cell carcinoma(ESCC) remains elusive.This study was conducted to investigate the potential effect and underlying molecular mechanism of melatonin as single anticancer agent against ESCC cells.Methods:ESCC cell lines treated with or without melatonin were used in this study.In vitro colony formation and 5-Ethynyl-2’-deoxyuridine(EdU) incorporation assays,and nude mice tumor xenograft model were used to confirm the proliferative capacities of ESCC cells.RNA-seq,qPCR,Western blotting,recombinant lentivirus-mediated target gene overexpression or knockdown,plasmids transfection and co-IP were applied to investigate the underlying molecular mechanism by which melatonin inhibited ESCC cell growth.IHC staining on ESCC tissue microarray and further survival analyses were performed to explore the relationship between target genes’ expression and prognosis of ESCC.Results:Melatonin treatment dose-dependently inhibited the proliferative ability and the expression of histone deacetylase 7(HDAC7),c-Myc and ubiquitin-specific peptidase 10(USP10) in ESCC cells(P<0.05).The expressions of HDAC7,c-Myc and USP10 in tumors were significantly higher than the paired normal tissues from 148 ESCC patients(P<0.001).Then,the Kaplan-Meier survival analysis suggested that ESCC patients with high HDAC7,c-Myc or USP10levels predicted worse overall survival(log-rank P<0.001).Co-IP and Western blotting further revealed that HDAC7physically deacetylated and activated β-catenin thus promoting downstream target c-Myc gene transcription.Notably,our mechanistic study validated that HDAC7/β-catenin/c-Myc could form the positive feedback loop to enhance ESCC cell growth,and USP10 could deubiquitinate and stabilize HDAC7 protein in the ESCC cells.Additionally,we verified that inhibition of the HDAC7/β-catenin/c-Myc axis and USP10/HDAC7 pathway mediated the anti-proliferative action of melatonin on ESCC cells.Conclusions:Our findings elucidate that melatonin mitigates the HDAC7/β-catenin/c-Myc positive feedback loop and inhibits the USP10-maintained HDAC7 protein stability thus suppressing ESCC cell growth,and provides the reference for identifying biomarkers and therapeutic targets for ESCC.
基金support from the National Natural Science Foundation of China(Nos.82173660,82103975)Zhejiang Provincial Key Research&Development Plan(No.2023C03111,China)+1 种基金the Natural Science Fund for Distinguished Young Scholars of Zhejiang Province(Nos.LR21H300003,LR22H310002,China)the Natural Science Foundation of Zhejiang Province(No.LQ21H300005,China).
文摘HDAC7,a member of class IIa HDACs,plays a pivotal regulatory role in tumor,immune,fibrosis,and angiogenesis,rendering it a potential therapeutic target.Nevertheless,due to the high similarity in the enzyme active sites of class IIa HDACs,inhibitors encounter challenges in discerning differences among them.Furthermore,the substitution of key residue in the active pocket of class IIa HDACs renders them pseudo-enzymes,leading to a limited impact of enzymatic inhibitors on their function.In this study,proteolysis targeting chimera(PROTAC)technology was employed to develop HDAC7 drugs.We developed an exceedingly selective HDAC7 PROTAC degrader B14 which showcased superior inhibitory effects on cell proliferation compared to TMP269 in various diffuse large B cell lymphoma(DLBCL)and acute myeloid leukemia(AML)cells.Subsequent investigations unveiled that B14 disrupts BCL6 forming a transcriptional inhibition complex by degrading HDAC7,thereby exerting proliferative inhibition in DLBCL.Our study broadened the understanding of the non-enzymatic functions of HDAC7 and underscored the importance of HDAC7 in the treatment of hematologic malignancies,particularly in DLBCL and AML.
基金supported by the National Heart,Lung,and Blood Institute R01 grant(HL153406).
文摘Postnatal mammalian cardiomyocytes(CMs)rapidly lose proliferative capacity and exit the cell cycle to undergo further differentiation and maturation.Cell cycle activation has been a major strategy to stimulate postnatal CM proliferation,albeit achieving modest effects.One impediment is that postnatal CMs may need to undergo dedifferentiation before proliferation,if not simultaneously.Here,we report that overexpression of Hdac7 in neonatal mouse CMs results in significant CM dedifferentiation and proliferation.Mechanistically,we showthat histone deacetylase7(HDAC7)-mediatedCM proliferation is contingent on dedifferentiation,which is accomplished by suppressing myocyte enhancefactor2(MEF2).Hdac7overexpression in CM shifts the chromatin state from binding with MEF2,which favors the transcriptional program toward differentiation,to binding with AP-1,which favors the transcriptional program toward proliferation.Furthermore,we found that HDAC7 interacts with minichromosome maintenance complex components to initiate cell cycleprogression.Ourfindings revealthat HDAC7 promotes CM proliferation byits dual action on CM dedifferentiation and proliferation,uncovering a potential new strategy for heart regeneration/repair.
文摘目的探讨白皮杉醇对脓毒症肺损伤小鼠代谢重编程的影响及可能机制。方法采用随机数字表法将32只雄性C57/B6小鼠随机分为4组,即对照组、模型组、低剂量治疗组和高剂量治疗组,每组各8只。对照组不做任何处理,其他3组使用脂多糖(lipopolysaccharide,LPS,10mg/kg)单次腹腔注射的方式构建脓毒症肺损伤小鼠模型,其中低剂量治疗组和高剂量治疗组小鼠于LPS注射前7天分别给予25mg/(kg·d)和100mg/(kg·d)的白皮杉醇灌胃,对照组和模型组给予等量0.9%氯化钠溶液灌胃。LPS腹腔注射12h后,留取小鼠支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)和肺组织,通过形态学和分子生物学实验评估各组小鼠肺损伤状况。结果与对照组比较,模型组小鼠肺组织病理损伤较对照组更明显,主要表现为肺水肿和炎性细胞浸润,肺损伤评分更高(8.25±0.45分vs 1.31±0.23分),BALF中乳酸含量(0.65±0.12mmol/L vs 0.17±0.04mmol/L)和乳酸盐脱氢酶活性(8934.12±145.25U/L vs 4782.53±111.04U/L)增加,肺组织中IL-1β、IL-6和TNF-α的mRNA水平明显升高,肺组织中糖酵解标志蛋白(GLUT1、HK2、PDK2、PKM2)和组蛋白去乙酰化酶7(histone deacetylase 7,HDAC7)的蛋白表达明显上调,差异均有统计学意义(P<0.05);与模型组比较,低剂量治疗组和高剂量治疗组小鼠肺病理损伤明显减轻,肺损伤评分更低(5.69±0.92分、2.04±0.31分vs 8.25±0.45分),BALF中乳酸含量(0.42±0.03mmol/L、0.25±0.02mmol/L vs 0.65±0.12mmol/L)和乳酸盐脱氢酶活性(6822.24±172.02U/L、5872.02±93.07U/L vs 8934.12±145.25U/L)降低,肺组织中IL-1β、IL-6和TNF-α的mRNA水平明显降低,肺组织中GLUT1、HK2、PDK2、PKM2和HDAC7的蛋白表达均明显下降,差异均有统计学意义(P<0.05)。结论白皮杉醇可显著减轻脓毒症小鼠肺病理损伤和炎性反应,其机制可能与HDAC7介导的代谢重编程抑制有关。
基金Relevant to this study,Chandra J has received research support from Karus Pharmaceuticals and Novartis PharmaceuticalsOrlowski RZ would like to acknowledge support from the National Cancer Institute(R01s CA184464 and CA194264)the Leukemia&Lymphoma Society Specialized Center of Research(SCOR-12206-17),and Dr.Miriam and Sheldon G.Adelson Research Foundation.
文摘Aim:Multiple myeloma(MM)is a hematological malignancy of antibody-producing mature B cells or plasma cells.The proteasome inhibitor,bortezomib,was the first-in-class compound to be FDA approved for MM and is frequently utilized in induction therapy.However,bortezomib refractory disease is a major clinical concern,and the efficacy of the pan-histone deacetylase inhibitor(HDACi),panobinostat,in bortezomib refractory disease indicates that HDAC targeting is a viable strategy.Here,we utilized isogenic bortezomib resistant models to profile HDAC expression and define baseline and HDACi-induced expression patterns of individual HDAC family members in sensitive vs.resistant cells to better understanding the potential for targeting these enzymes.Methods:Gene expression of HDAC family members in two sets of isogenic bortezomib sensitive or resistant myeloma cell lines was examined.These cell lines were subsequently treated with HDAC inhibitors:panobinostat or vorinostat,and HDAC expression was evaluated.CRISPR/Cas9 knockdown and pharmacological inhibition of specific HDAC family members were conducted.Results:Interestingly,HDAC6 and HDAC7 were significantly upregulated and downregulated,respectively,in bortezomib-resistant cells.Panobinostat was effective at inducing cell death in these lines and modulated HDAC expression in cell lines and patient samples.Knockdown of HDAC7 inhibited cell growth while pharmacologically inhibiting HDAC6 augmented cell death by panobinostat.Conclusion:Our data revealed heterogeneous expression of individual HDACs in bortezomib sensitive vs.resistant isogenic cell lines and patient samples treated with panobinostat.Cumulatively our findings highlight distinct roles for HDAC6 and HDAC7 in regulating cell death in the context of bortezomib resistance.
