Intervertebral disc degeneration(IDD)is a common chronic inflammatory degenerative disease that causes lower back pain.However,the underlying mechanisms of IDD remain unclear.Ferroptosis suppressor protein 1(FSP1)is a...Intervertebral disc degeneration(IDD)is a common chronic inflammatory degenerative disease that causes lower back pain.However,the underlying mechanisms of IDD remain unclear.Ferroptosis suppressor protein 1(FSP1)is a newly identified suppressor for ferroptosis.This study aims to investigate the role of FSP1 in IDD.Nucleus pulposus(NP)tissues in humans were collected and NP cells from rats were isolated to detect FSP1 expression pattern.The relationship between FSP1-mediated ferroptosis and apoptosis was identified using FSP1 inhibitor iFSP1.RNA sequencing was utilized to seek downstream molecules and related signaling pathways.Moreover,both exogenous recombinant FSP1 protein and endogenous small interfering RNA were implemented in this study to clarify the role of FSP1 in tumor necrosis factor-alpha(TNFα)-mediated NP cell apoptosis.Ultimately,the underlying mechanisms of FSP1-related signaling pathway in IDD were uncovered both in vitro and in vivo.As a result,FSP1 was up-regulated in human degenerative NP tissues and after TNFαstimulation.FSP1 inhibition by iFSP1 fails to trigger ferroptosis in NP cells while inhibiting TNFα-mediated apoptosis.Further experiments demonstrated that FSP1 was closely related to TNFα-reliant caspase 3 activation and mitochondrial damage.However,the exogenous addition of recombinant protein FSP1 does not induce cell death or intensify the efficacy of TNFα.Mechanically,FSP1 is involved in TNFα-mediated NF-κB signaling activation to accelerate the development of IDD.This study demonstrated that FSP1 promotes IDD through TNFα-reliant NF-κB signaling activation and caspase 3-dependent apoptosis.These findings suggested a novel therapeutic target for the treatment of IDD.展开更多
Doxorubicin(DOX)-mediated cardiotoxicity can exacerbate mortality in oncology patients,but related pharmacotherapeutic measures are relatively limited.Ferroptosis was recently identified as a major mechanism of DOX-in...Doxorubicin(DOX)-mediated cardiotoxicity can exacerbate mortality in oncology patients,but related pharmacotherapeutic measures are relatively limited.Ferroptosis was recently identified as a major mechanism of DOX-induced cardiotoxicity.Idebenone,a novel ferroptosis inhibitor,is a well-described clinical drug widely used.However,its role and pathological mechanism in DOX-induced cardiotoxicity are still unclear.In this study,we demonstrated the effects of idebenone on DOX-induced cardiotoxicity and elucidated its underlying mechanism.A single intraperitoneal injection of DOX(15 mg/kg)was administrated to establish DOX-induced cardiotoxicity.The results showed that idebenone significantly attenuated DOX-induced cardiac dysfunction due to its ability to regulate acute DOX-induced Fe^(2+)and ROS overload,which resulted in ferroptosis.CESTA and BLI further revealed that idebenone's anti-ferroptosis effect was mediated by FSP1.Interestingly,idebenone increased FSP1 protein levels but did not affect Fsp1 mRNA levels in the presence of DOX.Idebenone could form stable hydrogen bonds with FSP1 protein at K355,which may influence its association with ubiquitin.The results confirmed that idebenone stabilized FSP1 protein levels by inhibiting its ubiquitination degradation.In conclusion,this study demonstrates idebenone attenuated DOX-induced cardiotoxicity by inhibiting ferroptosis via regulation of FSP1,making it a potential clinical drug for patients receiving DOX treatment.展开更多
Reactive oxidative species(ROos)production-driven ferroptosis plays a role in acute kidney injury(Akl).However,its exact molecular mechanism is poorly understood.Scavenger receptor CD36 has important roles in oxidizin...Reactive oxidative species(ROos)production-driven ferroptosis plays a role in acute kidney injury(Akl).However,its exact molecular mechanism is poorly understood.Scavenger receptor CD36 has important roles in oxidizing lipids,lipid accumulation,metabolic syndrome,and insulin resistance in chronic kidney disease,but its roles remain unexplored in AKl.The present study investigated the role and mechanism of CD36 in regulating proximal tubular cell ferroptosis and AKl.The expression of CD36 was found to be significantly up-regulated in AKI renal tissues and correlated with renal function,which might serve as an independent biomarker for AKl patients.Moreover,in adult mice subjected to AKl,deletion of CD36(CD36-/-)induced tubular cell Ros accumulation,ferroptosis activation,and renal injury.Mechanistically,combining LC-MS/MS,co-IP,and ubiquitination analyses revealed that CD36 could specifically bind to ferroptosis suppressor protein 1(FSP1)and regulate its ubiquitination at sites K16 and K24,leading to FSP1 degradation and progression of ferroptosis in AKl.The present results emphasize a novel mechanism of CD36 in cisplatin-induced AKl.The discovery of the special CD36 roles in promoting ferroptosis and AKI development by regulating the ubiquitination of FSP1 in proximal tubular cells may be potential therapeutic targets for AKl.Moreover,CD36 may play a key role in the progression of AKl.Therefore,targeting CD36 may provide a promising treatment option for AKI.展开更多
Ferroptosis is a non-apoptotic mode of cell death characterized by iron-dependent accumulation of lipid peroxidation.While lipid radical elimination reaction catalyzed by glutathione peroxidase 4(GPX4)is a major anti-...Ferroptosis is a non-apoptotic mode of cell death characterized by iron-dependent accumulation of lipid peroxidation.While lipid radical elimination reaction catalyzed by glutathione peroxidase 4(GPX4)is a major anti-ferroptosis mechanism,inhibiting this pathway pharmaceutically shows promise as an antitumor strategy.However,certain tumor cells exhibit redundancy in lipid radical elimination pathways,rendering them unresponsive to GPX4 inhibitors.In this study,we conducted screens across different cancer cell lines and Food and Drug Administration-approved drugs,leading to the identification of temsirolimus in combination with the GPX4 inhibitor RSL3 as a potent inducer of ferroptosis in liver cancer cells.Mechanistically,temsirolimus sensitized liver cancer cells to ferroptosis by directly binding to and inhibiting ferroptosis suppressor protein 1(FSP1)enzyme.Notably,while temsirolimus is recognized as a potent mammalian target of rapamycin(mTOR)inhibitor,its ferroptosis-inducing effect is primarily attributed to the inhibition of FSP1 rather than mTOR activity.By employing in vitro colony formation assays and in vivo tumor xenograft models,we demonstrated that the combination of temsirolimus and RSL3 effectively suppressed liver tumor progression.This tumoricidal effect was associated with increased lipid peroxidation and induction of ferroptosis.In conclusion,our findings underscore the potential of combining multitarget ferroptosis-inducing agents to circumvent the resistance to ferroptosis of liver cancer cells and highlight temsirolimus as a promising FSP1 inhibitor and ferroptosis inducer,which also deserves further investigation in translational medicine.展开更多
Background:N-acetyltransferase 10(NAT10)is the only enzyme known tomediate the N4-acetylcytidine(ac4C)modification of mRNA and is crucial formRNA stability and translation efficiency.However,its role in cancer develop...Background:N-acetyltransferase 10(NAT10)is the only enzyme known tomediate the N4-acetylcytidine(ac4C)modification of mRNA and is crucial formRNA stability and translation efficiency.However,its role in cancer development and prognosis has not yet been explored.This study aimed to examine the possible role of NAT10 in colon cancer.Methods:The expression levels ofNAT10were evaluated by immunohistochemical analyses with a colon cancer tissue microarray,and its prognostic value in patients was further analyzed.Quantitative real-time polymerase chain reaction(qRT-PCR)and Western blotting were performed to analyze NAT10 expression in harvested colon cancer tissues and cell lines.Stable NAT10-knockdown and NAT10-overexpressing colon cancer cell lines were constructed using lentivirus.The biological functions of NAT10 in colon cancer cell lines were analyzed in vitro by Cell Counting Kit-8(CCK-8),wound healing,Transwell,cell cycle,and ferroptosis assays.Xenograft models were used to analyze the effect of NAT10 on the tumorigenesis and metastasis of colon cancer cells in vivo.Dot blotting,acetylated RNA immunoprecipitation-qPCR,and RNA stability analyses were performed to explore the mechanism by which NAT10 functions in colon cancer progression.Results:NAT10 was upregulated in colon cancer tissues and various colon cancer cell lines.This increased NAT10 expression was associated with shorter patient survival.Knockdown of NAT10 in two colon cancer cell lines(HT-29 and LoVo)impaired the proliferation,migration,invasion,tumor formation and metastasis of these cells,whereas overexpression of NAT10 promoted these abilities.Further analysis revealed that NAT10 exerted a strong effect on the mRNA stability and expression of ferroptosis suppressor protein 1(FSP1)in HT-29 and LoVo cells.In these cells,FSP1 mRNA was found to be modified by ac4C acetylation,and this epigenetic modification was associated with the inhibition of ferroptosis.Conclusions:Our study revealed that NAT10 plays a critical role in colon cancer development by affecting FSP1 mRNA stability and ferroptosis,suggesting that NAT10 could be a novel prognostic and therapeutic target in colon cancer.展开更多
基金supported in part by the National Natural Science Foundation of China(No.81874022 and 82172483 to Xinyu Liu,No.82102522 to Lianlei Wang)Key R&D Project of Shandong Province(China)(No.2022CXGC010503 to Xinyu Liu)+2 种基金Shandong Natural Science Foundation(No.ZR202102210113 to Lianlei Wang)Shandong Province Taishan Scholar Project(No.tsqn 202211317 to Lianlei Wang)National High Level Hospital Clinical Research Funding(No.2022-PUMCH-D-004).
文摘Intervertebral disc degeneration(IDD)is a common chronic inflammatory degenerative disease that causes lower back pain.However,the underlying mechanisms of IDD remain unclear.Ferroptosis suppressor protein 1(FSP1)is a newly identified suppressor for ferroptosis.This study aims to investigate the role of FSP1 in IDD.Nucleus pulposus(NP)tissues in humans were collected and NP cells from rats were isolated to detect FSP1 expression pattern.The relationship between FSP1-mediated ferroptosis and apoptosis was identified using FSP1 inhibitor iFSP1.RNA sequencing was utilized to seek downstream molecules and related signaling pathways.Moreover,both exogenous recombinant FSP1 protein and endogenous small interfering RNA were implemented in this study to clarify the role of FSP1 in tumor necrosis factor-alpha(TNFα)-mediated NP cell apoptosis.Ultimately,the underlying mechanisms of FSP1-related signaling pathway in IDD were uncovered both in vitro and in vivo.As a result,FSP1 was up-regulated in human degenerative NP tissues and after TNFαstimulation.FSP1 inhibition by iFSP1 fails to trigger ferroptosis in NP cells while inhibiting TNFα-mediated apoptosis.Further experiments demonstrated that FSP1 was closely related to TNFα-reliant caspase 3 activation and mitochondrial damage.However,the exogenous addition of recombinant protein FSP1 does not induce cell death or intensify the efficacy of TNFα.Mechanically,FSP1 is involved in TNFα-mediated NF-κB signaling activation to accelerate the development of IDD.This study demonstrated that FSP1 promotes IDD through TNFα-reliant NF-κB signaling activation and caspase 3-dependent apoptosis.These findings suggested a novel therapeutic target for the treatment of IDD.
基金supported by the following grants:the Regional Innovation and Development Joint Fund of the National Natural Science Foundation of China(No.U22A20269)the National Key R&D Program of China(No.2018YFC1311300)the National Natural Science Foundation of China(Nos.82200262 and 82000229).
文摘Doxorubicin(DOX)-mediated cardiotoxicity can exacerbate mortality in oncology patients,but related pharmacotherapeutic measures are relatively limited.Ferroptosis was recently identified as a major mechanism of DOX-induced cardiotoxicity.Idebenone,a novel ferroptosis inhibitor,is a well-described clinical drug widely used.However,its role and pathological mechanism in DOX-induced cardiotoxicity are still unclear.In this study,we demonstrated the effects of idebenone on DOX-induced cardiotoxicity and elucidated its underlying mechanism.A single intraperitoneal injection of DOX(15 mg/kg)was administrated to establish DOX-induced cardiotoxicity.The results showed that idebenone significantly attenuated DOX-induced cardiac dysfunction due to its ability to regulate acute DOX-induced Fe^(2+)and ROS overload,which resulted in ferroptosis.CESTA and BLI further revealed that idebenone's anti-ferroptosis effect was mediated by FSP1.Interestingly,idebenone increased FSP1 protein levels but did not affect Fsp1 mRNA levels in the presence of DOX.Idebenone could form stable hydrogen bonds with FSP1 protein at K355,which may influence its association with ubiquitin.The results confirmed that idebenone stabilized FSP1 protein levels by inhibiting its ubiquitination degradation.In conclusion,this study demonstrates idebenone attenuated DOX-induced cardiotoxicity by inhibiting ferroptosis via regulation of FSP1,making it a potential clinical drug for patients receiving DOX treatment.
基金supported by grants from the National Natural Science Foundation of China(No.81873604)the Medical Scientific Research Project of the Chongqing Health Commission(China)(No.2022GDRC005)Chongqing Science and Technology Agency(China)(CSTB2022NSCQ-MSX0984).
文摘Reactive oxidative species(ROos)production-driven ferroptosis plays a role in acute kidney injury(Akl).However,its exact molecular mechanism is poorly understood.Scavenger receptor CD36 has important roles in oxidizing lipids,lipid accumulation,metabolic syndrome,and insulin resistance in chronic kidney disease,but its roles remain unexplored in AKl.The present study investigated the role and mechanism of CD36 in regulating proximal tubular cell ferroptosis and AKl.The expression of CD36 was found to be significantly up-regulated in AKI renal tissues and correlated with renal function,which might serve as an independent biomarker for AKl patients.Moreover,in adult mice subjected to AKl,deletion of CD36(CD36-/-)induced tubular cell Ros accumulation,ferroptosis activation,and renal injury.Mechanistically,combining LC-MS/MS,co-IP,and ubiquitination analyses revealed that CD36 could specifically bind to ferroptosis suppressor protein 1(FSP1)and regulate its ubiquitination at sites K16 and K24,leading to FSP1 degradation and progression of ferroptosis in AKl.The present results emphasize a novel mechanism of CD36 in cisplatin-induced AKl.The discovery of the special CD36 roles in promoting ferroptosis and AKI development by regulating the ubiquitination of FSP1 in proximal tubular cells may be potential therapeutic targets for AKl.Moreover,CD36 may play a key role in the progression of AKl.Therefore,targeting CD36 may provide a promising treatment option for AKI.
基金funded by grants from the National Key R&D Program of China(2020YFA0803202 to D.Y.)the National Natural Science Foundation of China(82225036 and 31821002 to D.Y.,32101011 to P.W.).
文摘Ferroptosis is a non-apoptotic mode of cell death characterized by iron-dependent accumulation of lipid peroxidation.While lipid radical elimination reaction catalyzed by glutathione peroxidase 4(GPX4)is a major anti-ferroptosis mechanism,inhibiting this pathway pharmaceutically shows promise as an antitumor strategy.However,certain tumor cells exhibit redundancy in lipid radical elimination pathways,rendering them unresponsive to GPX4 inhibitors.In this study,we conducted screens across different cancer cell lines and Food and Drug Administration-approved drugs,leading to the identification of temsirolimus in combination with the GPX4 inhibitor RSL3 as a potent inducer of ferroptosis in liver cancer cells.Mechanistically,temsirolimus sensitized liver cancer cells to ferroptosis by directly binding to and inhibiting ferroptosis suppressor protein 1(FSP1)enzyme.Notably,while temsirolimus is recognized as a potent mammalian target of rapamycin(mTOR)inhibitor,its ferroptosis-inducing effect is primarily attributed to the inhibition of FSP1 rather than mTOR activity.By employing in vitro colony formation assays and in vivo tumor xenograft models,we demonstrated that the combination of temsirolimus and RSL3 effectively suppressed liver tumor progression.This tumoricidal effect was associated with increased lipid peroxidation and induction of ferroptosis.In conclusion,our findings underscore the potential of combining multitarget ferroptosis-inducing agents to circumvent the resistance to ferroptosis of liver cancer cells and highlight temsirolimus as a promising FSP1 inhibitor and ferroptosis inducer,which also deserves further investigation in translational medicine.
基金National Natural Science Foundation of China,Grant/Award Numbers:81902386,81972869,82002479The Natural Science Foundation of Jiangsu Province,Grant/Award Numbers:BK20211065,BK20200179+2 种基金China Postdoctoral Science Foundation,Grant/Award Number:2021M700547Youth Talent Science and Technology Project of Changzhou Health Commission,Grant/Award Number:QN202103The open fund of state key laboratory of Pharmaceutical Biotechnology,Nanjing University,China,Grant/Award Number:KF-202203。
文摘Background:N-acetyltransferase 10(NAT10)is the only enzyme known tomediate the N4-acetylcytidine(ac4C)modification of mRNA and is crucial formRNA stability and translation efficiency.However,its role in cancer development and prognosis has not yet been explored.This study aimed to examine the possible role of NAT10 in colon cancer.Methods:The expression levels ofNAT10were evaluated by immunohistochemical analyses with a colon cancer tissue microarray,and its prognostic value in patients was further analyzed.Quantitative real-time polymerase chain reaction(qRT-PCR)and Western blotting were performed to analyze NAT10 expression in harvested colon cancer tissues and cell lines.Stable NAT10-knockdown and NAT10-overexpressing colon cancer cell lines were constructed using lentivirus.The biological functions of NAT10 in colon cancer cell lines were analyzed in vitro by Cell Counting Kit-8(CCK-8),wound healing,Transwell,cell cycle,and ferroptosis assays.Xenograft models were used to analyze the effect of NAT10 on the tumorigenesis and metastasis of colon cancer cells in vivo.Dot blotting,acetylated RNA immunoprecipitation-qPCR,and RNA stability analyses were performed to explore the mechanism by which NAT10 functions in colon cancer progression.Results:NAT10 was upregulated in colon cancer tissues and various colon cancer cell lines.This increased NAT10 expression was associated with shorter patient survival.Knockdown of NAT10 in two colon cancer cell lines(HT-29 and LoVo)impaired the proliferation,migration,invasion,tumor formation and metastasis of these cells,whereas overexpression of NAT10 promoted these abilities.Further analysis revealed that NAT10 exerted a strong effect on the mRNA stability and expression of ferroptosis suppressor protein 1(FSP1)in HT-29 and LoVo cells.In these cells,FSP1 mRNA was found to be modified by ac4C acetylation,and this epigenetic modification was associated with the inhibition of ferroptosis.Conclusions:Our study revealed that NAT10 plays a critical role in colon cancer development by affecting FSP1 mRNA stability and ferroptosis,suggesting that NAT10 could be a novel prognostic and therapeutic target in colon cancer.
