Mevalonate pathway for isoprenoid biosynthesis was constructed in Escherichia coli cells by the transformation with a gene cluster isolated from Streptomyces sp., and farnesyl diphosphate synthase and δ-guaiene synth...Mevalonate pathway for isoprenoid biosynthesis was constructed in Escherichia coli cells by the transformation with a gene cluster isolated from Streptomyces sp., and farnesyl diphosphate synthase and δ-guaiene synthase genes were coexpressed in this strain. This transformant was capable of liberating an appreciable amount of δ-guaiene, an aroma sesquiterpene compound accumulated in agarwood, and its concentration was elevated to more than 30 μg/ml culture by the incubation with mevalonolactone as an isoprene precursor in a nutrient-enriched Terrific broth. Coexpression of type 1 isopentenyl diphosphate isomerase plus acetoacetyl-CoA ligase genes also enhanced δ-guaiene production, and the concentration of the compound was approximately 38 - 42 μg/ml culture in the presence of mevalonolactone or lithium acetoacetate. These results clearly indicate that mevalonate pathway-engineered E. coli cells showed an appreciable δ-guaiene producing activity in the en- riched medium in the presence of appropriate isoprene precursors.展开更多
Zeaxanthin,an oxygenated carotenoid derivative with potent antioxidative properties,is produced by many organism taxa.Flavobacteriaceae are widely distributed in marine environments;however,the zeaxanthin biosynthesis...Zeaxanthin,an oxygenated carotenoid derivative with potent antioxidative properties,is produced by many organism taxa.Flavobacteriaceae are widely distributed in marine environments;however,the zeaxanthin biosynthesis property in this family remains incompletely explored.Here,we characterized zeaxanthin production by marine Flavobacteriaceae strains and elucidated underlying molecular mechanisms.Eight Flavobacteriaceae strains were isolated from the phycosphere of various dinoflagellates.Analyses of the zeaxanthin production in these strains revealed yields ranging from 5 to 3289μg/g of dry cell weight.Genomic and molecular biology analyses revealed the biosynthesized zeaxanthin through the mevalonate(MVA)pathway diverging from the 2-C-methyl-d-erythritol-4-phosphate(MEP)pathway commonly observed in most Gram-negative bacteria.Furthermore,comprehensive genome analyses of 322 culturable marine Flavobacteriale strains indicated that the majority of Flavobacteriaceae members possess the potential to synthesize zeaxanthin using precursors derived from the MVA pathway.These data provide insight into the zeaxanthin biosynthesis property in marine Flavobacteriaceae strains,highlighting their ecological and biotechnological relevance.展开更多
Osteosarcoma is the most common malignant bone tumour,and the metastasis of osteosarcoma is an important cause of death.Evidence has shown that the mevalonate pathway is highly activated and is expected to be a new ta...Osteosarcoma is the most common malignant bone tumour,and the metastasis of osteosarcoma is an important cause of death.Evidence has shown that the mevalonate pathway is highly activated and is expected to be a new target for tumour therapy.In this study,we investigated the effect of mevalonate signalling on osteosarcoma metastasis and its molecular mechanism.First,we found that the key rate-limiting enzyme of mevalonate signalling,3-hydroxy-3-methylglutaryl-CoA reductase(HMGCR),was highly expressed in osteosarcoma cells,and inhibition of HMGCR with simvastatin significantly inhibited the motility of 143B cells.Next,we found that YAP1 activity was significantly upregulated in osteosarcoma cells and that YAP1 knockdown inhibited the motility of 143B cells.We also found that the mevalonate pathway regulated the motility of 143B cells by modulating YAP1 phosphorylation and cellular localization.Moreover,we found that the activity of YAP1 was regulated by the mevalonate pathway by modulating the cell membrane localization of RhoA.Finally,we demonstrated that inhibition of the mevalonate pathway notably reduced the lung metastasis of 143B cells,as reflected by the decreased incidence and number of metastatic nodules and the increased survival time of the nude mice.Taken together,our findings suggest that the mevalonate pathway can promote the metastasis of osteosarcoma by activating YAP1 via RhoA.Inhibition of the mevalonate pathway may be a promising therapeutic strategy for osteosarcoma metastasis.展开更多
Dysregulation of mevalonate pathway,an essential metabolic route involving coenzyme A(CoASH)and cholesterol,contributes significantly to escalating cartilage degradation.Existing treatments rely on the sim-vastatin de...Dysregulation of mevalonate pathway,an essential metabolic route involving coenzyme A(CoASH)and cholesterol,contributes significantly to escalating cartilage degradation.Existing treatments rely on the sim-vastatin delivery via tunable sol-gel transition mechanisms of injectable hydrogel.However,those methods suffer from lack of controllable drug release by selective phase transition under distinct disease microenvironment.Herein,we developed an aberrant lipid metabolism microenvironment-activated phase transition(normal con-dition:gel-gel,abnormal condition:gel-sol)with targeted drug release for synergistic treatment of osteoarthritis(OA).Naked-eye diagnosis and therapy of OA through cholesterol downregulation using an injectable hydrogel were based on the simvastatin-loaded nanoparticles embedded in hexanoyl glycol chitosan(HGC-SIM@PAAMnO2-cPDA or SIM gel).The interaction between highly expressed CoASH in OA and PAA-MnO2 in SIM gel altered the hydrophobic-hydrophilic balance and gelation temperature,triggering the OA-sensitive gel-sol transformation.Naked-eye gel-sol transformation was observed after incubating SIM gel with OA chondrocyte models,including acetyl-CoA-induced wild-type(WT+CoA),NudT7^(-/-)knockout(N7KO),and Acot12^(-/-)knockout(A12KO).Because of the simvastatin release after gel-sol transition,OA-related enzymes and genes,including antioxidant enzymes(Sod2),cartilage degradation genes(Adamts4),and cholesterol synthesis-related enzymes(Mvk),were downregulated.In vivo studies revealed gel-sol transformation in destabilized medial meniscus of OA mice(DMM WT,N7KO,and A12KO)at 4-8 weeks post-injection,with significantly reduced cartilage degradation,demonstrating theragnostic capability of SIM gel.Thus,SIM gel offers a potential approach for future synergistic OA diagnosis and therapy.展开更多
TheγδT cells are an emerging class of immune effectors with potent antitumor activity,bridging innate and adaptive immunity.Their unique ability to recognise stress-induced ligands independently of major histocompat...TheγδT cells are an emerging class of immune effectors with potent antitumor activity,bridging innate and adaptive immunity.Their unique ability to recognise stress-induced ligands independently of major histocompatibility complex restriction makes them attractive candidates for cancer immunotherapy.However,the clinical application ofγδT cells requires efficient in vitro expansion strategies to generate large numbers of functional cells.This mini-review explores the latest advancements inγδT cell expansion protocols,focusing on key activation stimuli,cytokine support,and culture conditions that optimise proliferation and cytotoxicity.展开更多
Statins inhibit 3-hydroxy-3-methylglutaryl-CoA reductase,the rate-limiting enzyme of the mevalonate pathway,and are widely used as an effective and safe approach handle hypercholesterolemia.The mevalonate pathway is a...Statins inhibit 3-hydroxy-3-methylglutaryl-CoA reductase,the rate-limiting enzyme of the mevalonate pathway,and are widely used as an effective and safe approach handle hypercholesterolemia.The mevalonate pathway is a vital metabolic pathway that uses acetyl-CoA to generate isoprenoids and sterols that are crucial to tumor growth and progression.Multiple studies have indicated that statins improve patient prognosis in various carcinomas.Basic research on the mechanisms underlying the antitumor effects of statins is underway.The development of new anti-cancer drugs is progressing,but increasing medical costs from drug development have become a major obstacle.Readily available,inexpensive and well-tolerated drugs like statins have not yet been successfully repurposed for cancer treatment.Identifying the cancer patients that may benefit from statins is key to improved patient treatment.This review summarizes recent advances in statin research in cancer and suggests important considerations for the clinical use of statins to improve outcomes for cancer patients.展开更多
Statins,which are competitive inhibitors of 3-hydroxy-3-methyl-glutarylcoenzyme A reductase,reduce cholesterol blood levels and the risk of developing cardiovascular diseases and their related complications.In additio...Statins,which are competitive inhibitors of 3-hydroxy-3-methyl-glutarylcoenzyme A reductase,reduce cholesterol blood levels and the risk of developing cardiovascular diseases and their related complications.In addition to this main activity,statins show pleiotropic effects such as antioxidant,anti-inflammatory and antiproliferative properties,with applications in many pathologies.Based on their antiproliferative properties,in vitro and in vivo studies have investigated their effects on various types of cancer(i.e.,breast cancer,prostate cancer,colorectal cancer,ovarian cancer,lung cancer)with different genetic and molecular characteristics.Many positive results were obtained,but they were highly dependent on the physiochemical properties of the statins,their dose and treatment period.Combined therapies of statins and cytotoxic drugs have also been tested,and synergistic or additive effects were observed.Moreover,observational studies performed on patients who used statins for different pathologies,revealed that statins reduced the risk of developing various cancers,and improved the outcomes for cancer patients.Currently,there are many ongoing clinical trials aimed at exploring the potential of statins to lower the mortality and the disease-recurrence risk.All these results are the foundation of new treatment directions in cancer therapy.展开更多
The isoprenoid brasilicardin A is a promising immunosuppressant compound with a unique mode of action,high potency and reduced toxicity compared to today's standard drugs.However,production of brasilicardin has be...The isoprenoid brasilicardin A is a promising immunosuppressant compound with a unique mode of action,high potency and reduced toxicity compared to today's standard drugs.However,production of brasilicardin has been hampered since the producer strain Nocardia terpenica IFM0406 synthesizes brasilicardin in only low amounts and is a biosafety level 2 organism.Previously,we were able to heterologously express the brasilicardin gene cluster in the nocardioform actinomycete Amycolatopsis japonicum.Four brasilicardin congeners,intermediates of the BraA biosynthesis,were produced.Since chemical synthesis of the brasilicardin core structure has remained elusive we intended to produce high amounts of the brasilicardin backbone for semi synthesis and derivatization.Therefore,we used a metabolic engineering approach to increase heterologous production of brasilicardin in A.japonicum.Simultaneous heterologous expression of genes encoding the MVA pathway and expression of diterpenoid specific prenyltransferases were used to increase the provision of the isoprenoid precursor isopentenyl diphosphate(IPP)and to channel the precursor into the direction of diterpenoid biosynthesis.Both approaches contributed to an elevated heterologous production of the brasilicardin backbone,which can now be used as a starting point for semi synthesis of new brasilicardin congeners with better properties.展开更多
Being a Th2 stimulator,classic aluminum salt-based adjuvants only stimulate weak cellular immune responses that are required for vaccination against intracellular viruses or cancerous cells.As a thirdgeneration bispho...Being a Th2 stimulator,classic aluminum salt-based adjuvants only stimulate weak cellular immune responses that are required for vaccination against intracellular viruses or cancerous cells.As a thirdgeneration bisphosphonate,zoledronate(ZOL)can enhance antigen crosspresentation by inhibiting key enzymes of the mevalonate pathway.Here,we developed the subunit antigen ovalbumin(OVA)and ZOL co-loaded aluminum hydroxide nanoparticles(APN-OVA-ZOL)and investigated their capacity for inducing cellular immune responses against the antigen.Our results showed that the developed nanovaccines could successfully encapsulate OVA and ZOL,and enabled efficient lymph node delivery.Benefited by the mevalonate pathway inhibition effect of ZOL,APN-OVA-ZOL significantly promoted crosspresentation.As a result,APN-OVA-ZOL induced robust cellular immunity,including the activation of T and B cells.In a EG7-OVA tumor-bearing murine model,APN-OVA-ZOL significantly inhibited the tumor growth and prolonged mice survival.This work provided a strong empirical foundation indicating that zoledronate-loaded aluminum salt nanovaccines had a strong potency for cancer immunotherapy.展开更多
All organisms living in complex environments have evolved effective mechanisms of dynamic responses to extracellular stimuli.The immune system activates when damaged or injured cells release damage‐associated molecul...All organisms living in complex environments have evolved effective mechanisms of dynamic responses to extracellular stimuli.The immune system activates when damaged or injured cells release damage‐associated molecular patterns(DAMPs).In addition to well‐characterized DAMPs such as high‐mobility group box 1 and adenosine triphosphate,studies on new classes of DAMPs have emerged.Here,we review recent reports of a new class of isoprenoid‐derived DAMPs,including farnesyl pyrophosphate and geranylgeranyl pyrophosphate,both of which are pivotal metabolic inter-mediates of the mevalonate pathway.We also explore the roles of old and new DAMPs in autoimmune diseases that result from dysregulated inflammation.The findings highlight that understanding the functional mechanisms of DAMPs is important to enrich the DAMP family and decipher their immunoregulatory mechanisms to provide new therapeutics for the prevention and treatment of autoimmune diseases.展开更多
Leukemia is a common hematological malignancy with overall poor prognosis.Novel therapies are needed to improve the outcome of leukemia patients.Cholesterol metabolism reprogramming is a featured alteration in leukemi...Leukemia is a common hematological malignancy with overall poor prognosis.Novel therapies are needed to improve the outcome of leukemia patients.Cholesterol metabolism reprogramming is a featured alteration in leukemia.Many metabolic-related genes and metabolites are essential to the progress and drug resistance of leukemia.Exploring potential therapeutical targets related to cholesterol homeostasis is a promising area.This review summarized the functions of cholesterol and its derived intermediate metabolites,and also discussed potential agents targeting this metabolic vulnerability in leukemia.展开更多
Aim: This study aimed to decipher the molecular mechanism underlying the synergistic effect of inhibitors of the mevalonate-cholesterol pathway (i.e., statins) and aminopeptidase inhibitors (APis) on APi-sensitive and...Aim: This study aimed to decipher the molecular mechanism underlying the synergistic effect of inhibitors of the mevalonate-cholesterol pathway (i.e., statins) and aminopeptidase inhibitors (APis) on APi-sensitive and -resistant acute myeloid leukemia (AML) cells.Methods: U937 cells and their sublines with low and high levels of acquired resistance to (6S)-[(R)-2-((S)-Hydroxy-hydroxycarbamoyl-methoxy-methyl)-4-methyl-pentanoylamino]-3,3 dimethyl-butyric acid cyclopentyl ester (CHR2863), an APi prodrug, served as main AML cell line models. Drug combination effects were assessed with CHR2863 and in vitro non-toxic concentrations of various statins upon cell growth inhibition, cell cycle effects, and apoptosis induction. Mechanistic studies involved analysis of Rheb prenylation required for mTOR activation.Results: A strong synergy of CHR2863 with the statins simvastatin, fluvastatin, lovastatin, and pravastatin was demonstrated in U937 cells and two CHR2863-resistant sublines. This potent synergy between simvastatin and CHR2863 was also observed with a series of other human AML cell lines (e.g., THP1, MV4-11, and KG1), but not with acute lymphocytic leukemia or multiple solid tumor cell lines. This synergistic activity was: (i) specific for APis (e.g., CHR2863 and Bestatin), rather than for other cytotoxic agents;and (ii) corroborated by enhanced induction of apoptosis and cell cycle arrest which increased the sub-G1 fraction. Consistently, statin potentiation of CHR2863 activity was abrogated by co-administration of mevalonate and/or farnesyl pyrophosphate, suggesting the involvement of protein prenylation;this was experimentally confirmed by impaired Rheb prenylation by simvastatin.Conclusion: These novel findings suggest that the combined inhibitory effect of impaired Rheb prenylation and CHR2863-dependent mTOR inhibition instigates a potent synergistic inhibition of statins and APis on human AML cells.展开更多
Cycloastragenol is a bioactive,high-value triterpenoid derived from Astragalus membranaceus.Conventional plant-based extraction and chemical synthesis methods are expensive.To our knowledge,this is the first report on...Cycloastragenol is a bioactive,high-value triterpenoid derived from Astragalus membranaceus.Conventional plant-based extraction and chemical synthesis methods are expensive.To our knowledge,this is the first report on the de novo biosynthesis of cycloastragenol in yeast.The mevalonate pathway was reconstituted in yeast peroxisomes,and the engineered yeast produced 656.55 mg/L squalene.Further introduction of heterologous enzymes led the engineered yeast to produce 1.04 mg/L cycloastragenol,which demonstrated the yeast production of value-added medicinal molecules.展开更多
文摘Mevalonate pathway for isoprenoid biosynthesis was constructed in Escherichia coli cells by the transformation with a gene cluster isolated from Streptomyces sp., and farnesyl diphosphate synthase and δ-guaiene synthase genes were coexpressed in this strain. This transformant was capable of liberating an appreciable amount of δ-guaiene, an aroma sesquiterpene compound accumulated in agarwood, and its concentration was elevated to more than 30 μg/ml culture by the incubation with mevalonolactone as an isoprene precursor in a nutrient-enriched Terrific broth. Coexpression of type 1 isopentenyl diphosphate isomerase plus acetoacetyl-CoA ligase genes also enhanced δ-guaiene production, and the concentration of the compound was approximately 38 - 42 μg/ml culture in the presence of mevalonolactone or lithium acetoacetate. These results clearly indicate that mevalonate pathway-engineered E. coli cells showed an appreciable δ-guaiene producing activity in the en- riched medium in the presence of appropriate isoprene precursors.
基金provided by the Guangdong Basic and Applied Basic Research Foundation(2022B1515020017,2021A1515110426)the National Natural Science Foundation of China(32070113).
文摘Zeaxanthin,an oxygenated carotenoid derivative with potent antioxidative properties,is produced by many organism taxa.Flavobacteriaceae are widely distributed in marine environments;however,the zeaxanthin biosynthesis property in this family remains incompletely explored.Here,we characterized zeaxanthin production by marine Flavobacteriaceae strains and elucidated underlying molecular mechanisms.Eight Flavobacteriaceae strains were isolated from the phycosphere of various dinoflagellates.Analyses of the zeaxanthin production in these strains revealed yields ranging from 5 to 3289μg/g of dry cell weight.Genomic and molecular biology analyses revealed the biosynthesized zeaxanthin through the mevalonate(MVA)pathway diverging from the 2-C-methyl-d-erythritol-4-phosphate(MEP)pathway commonly observed in most Gram-negative bacteria.Furthermore,comprehensive genome analyses of 322 culturable marine Flavobacteriale strains indicated that the majority of Flavobacteriaceae members possess the potential to synthesize zeaxanthin using precursors derived from the MVA pathway.These data provide insight into the zeaxanthin biosynthesis property in marine Flavobacteriaceae strains,highlighting their ecological and biotechnological relevance.
基金This work was supported by the Natural Science Foundation of Chongqing(No.cstc2019jcyj-msxmX0358).
文摘Osteosarcoma is the most common malignant bone tumour,and the metastasis of osteosarcoma is an important cause of death.Evidence has shown that the mevalonate pathway is highly activated and is expected to be a new target for tumour therapy.In this study,we investigated the effect of mevalonate signalling on osteosarcoma metastasis and its molecular mechanism.First,we found that the key rate-limiting enzyme of mevalonate signalling,3-hydroxy-3-methylglutaryl-CoA reductase(HMGCR),was highly expressed in osteosarcoma cells,and inhibition of HMGCR with simvastatin significantly inhibited the motility of 143B cells.Next,we found that YAP1 activity was significantly upregulated in osteosarcoma cells and that YAP1 knockdown inhibited the motility of 143B cells.We also found that the mevalonate pathway regulated the motility of 143B cells by modulating YAP1 phosphorylation and cellular localization.Moreover,we found that the activity of YAP1 was regulated by the mevalonate pathway by modulating the cell membrane localization of RhoA.Finally,we demonstrated that inhibition of the mevalonate pathway notably reduced the lung metastasis of 143B cells,as reflected by the decreased incidence and number of metastatic nodules and the increased survival time of the nude mice.Taken together,our findings suggest that the mevalonate pathway can promote the metastasis of osteosarcoma by activating YAP1 via RhoA.Inhibition of the mevalonate pathway may be a promising therapeutic strategy for osteosarcoma metastasis.
基金supported by National Research Foundation of Korea(NRF)grants funded by the Korean Government(MSIT)(No.RS-2023-00207925,RS-2023-00237700,2018R1A6A1A03023788 and RS-2024-00449612)the Alchemist Project of the Korea Evaluation Institute of Industrial Technology(KEIT 20018560,NTIS 1415184668)funded by the Ministry of Trade,Industry&Energy(MOTIE),Republic of Korea.
文摘Dysregulation of mevalonate pathway,an essential metabolic route involving coenzyme A(CoASH)and cholesterol,contributes significantly to escalating cartilage degradation.Existing treatments rely on the sim-vastatin delivery via tunable sol-gel transition mechanisms of injectable hydrogel.However,those methods suffer from lack of controllable drug release by selective phase transition under distinct disease microenvironment.Herein,we developed an aberrant lipid metabolism microenvironment-activated phase transition(normal con-dition:gel-gel,abnormal condition:gel-sol)with targeted drug release for synergistic treatment of osteoarthritis(OA).Naked-eye diagnosis and therapy of OA through cholesterol downregulation using an injectable hydrogel were based on the simvastatin-loaded nanoparticles embedded in hexanoyl glycol chitosan(HGC-SIM@PAAMnO2-cPDA or SIM gel).The interaction between highly expressed CoASH in OA and PAA-MnO2 in SIM gel altered the hydrophobic-hydrophilic balance and gelation temperature,triggering the OA-sensitive gel-sol transformation.Naked-eye gel-sol transformation was observed after incubating SIM gel with OA chondrocyte models,including acetyl-CoA-induced wild-type(WT+CoA),NudT7^(-/-)knockout(N7KO),and Acot12^(-/-)knockout(A12KO).Because of the simvastatin release after gel-sol transition,OA-related enzymes and genes,including antioxidant enzymes(Sod2),cartilage degradation genes(Adamts4),and cholesterol synthesis-related enzymes(Mvk),were downregulated.In vivo studies revealed gel-sol transformation in destabilized medial meniscus of OA mice(DMM WT,N7KO,and A12KO)at 4-8 weeks post-injection,with significantly reduced cartilage degradation,demonstrating theragnostic capability of SIM gel.Thus,SIM gel offers a potential approach for future synergistic OA diagnosis and therapy.
基金Supported by National Science Center of Poland,No.2019/35/N/NZ6/02973.
文摘TheγδT cells are an emerging class of immune effectors with potent antitumor activity,bridging innate and adaptive immunity.Their unique ability to recognise stress-induced ligands independently of major histocompatibility complex restriction makes them attractive candidates for cancer immunotherapy.However,the clinical application ofγδT cells requires efficient in vitro expansion strategies to generate large numbers of functional cells.This mini-review explores the latest advancements inγδT cell expansion protocols,focusing on key activation stimuli,cytokine support,and culture conditions that optimise proliferation and cytotoxicity.
文摘Statins inhibit 3-hydroxy-3-methylglutaryl-CoA reductase,the rate-limiting enzyme of the mevalonate pathway,and are widely used as an effective and safe approach handle hypercholesterolemia.The mevalonate pathway is a vital metabolic pathway that uses acetyl-CoA to generate isoprenoids and sterols that are crucial to tumor growth and progression.Multiple studies have indicated that statins improve patient prognosis in various carcinomas.Basic research on the mechanisms underlying the antitumor effects of statins is underway.The development of new anti-cancer drugs is progressing,but increasing medical costs from drug development have become a major obstacle.Readily available,inexpensive and well-tolerated drugs like statins have not yet been successfully repurposed for cancer treatment.Identifying the cancer patients that may benefit from statins is key to improved patient treatment.This review summarizes recent advances in statin research in cancer and suggests important considerations for the clinical use of statins to improve outcomes for cancer patients.
文摘Statins,which are competitive inhibitors of 3-hydroxy-3-methyl-glutarylcoenzyme A reductase,reduce cholesterol blood levels and the risk of developing cardiovascular diseases and their related complications.In addition to this main activity,statins show pleiotropic effects such as antioxidant,anti-inflammatory and antiproliferative properties,with applications in many pathologies.Based on their antiproliferative properties,in vitro and in vivo studies have investigated their effects on various types of cancer(i.e.,breast cancer,prostate cancer,colorectal cancer,ovarian cancer,lung cancer)with different genetic and molecular characteristics.Many positive results were obtained,but they were highly dependent on the physiochemical properties of the statins,their dose and treatment period.Combined therapies of statins and cytotoxic drugs have also been tested,and synergistic or additive effects were observed.Moreover,observational studies performed on patients who used statins for different pathologies,revealed that statins reduced the risk of developing various cancers,and improved the outcomes for cancer patients.Currently,there are many ongoing clinical trials aimed at exploring the potential of statins to lower the mortality and the disease-recurrence risk.All these results are the foundation of new treatment directions in cancer therapy.
基金This work was funded by Bundesministerium für Bildung und Forschung(BMBF)(FKZ 031A568B),in the frame of the ERA-NET-IB project“NeBrasCa”.
文摘The isoprenoid brasilicardin A is a promising immunosuppressant compound with a unique mode of action,high potency and reduced toxicity compared to today's standard drugs.However,production of brasilicardin has been hampered since the producer strain Nocardia terpenica IFM0406 synthesizes brasilicardin in only low amounts and is a biosafety level 2 organism.Previously,we were able to heterologously express the brasilicardin gene cluster in the nocardioform actinomycete Amycolatopsis japonicum.Four brasilicardin congeners,intermediates of the BraA biosynthesis,were produced.Since chemical synthesis of the brasilicardin core structure has remained elusive we intended to produce high amounts of the brasilicardin backbone for semi synthesis and derivatization.Therefore,we used a metabolic engineering approach to increase heterologous production of brasilicardin in A.japonicum.Simultaneous heterologous expression of genes encoding the MVA pathway and expression of diterpenoid specific prenyltransferases were used to increase the provision of the isoprenoid precursor isopentenyl diphosphate(IPP)and to channel the precursor into the direction of diterpenoid biosynthesis.Both approaches contributed to an elevated heterologous production of the brasilicardin backbone,which can now be used as a starting point for semi synthesis of new brasilicardin congeners with better properties.
基金supported by the National Natural Science Foundation of China(Nos.82320108020 and 81925036)the National Key Research and Development Program of China(No.2023YFC2307700)+1 种基金111 project(No.B18035)the Fundamental Research Funds for the Central Universities。
文摘Being a Th2 stimulator,classic aluminum salt-based adjuvants only stimulate weak cellular immune responses that are required for vaccination against intracellular viruses or cancerous cells.As a thirdgeneration bisphosphonate,zoledronate(ZOL)can enhance antigen crosspresentation by inhibiting key enzymes of the mevalonate pathway.Here,we developed the subunit antigen ovalbumin(OVA)and ZOL co-loaded aluminum hydroxide nanoparticles(APN-OVA-ZOL)and investigated their capacity for inducing cellular immune responses against the antigen.Our results showed that the developed nanovaccines could successfully encapsulate OVA and ZOL,and enabled efficient lymph node delivery.Benefited by the mevalonate pathway inhibition effect of ZOL,APN-OVA-ZOL significantly promoted crosspresentation.As a result,APN-OVA-ZOL induced robust cellular immunity,including the activation of T and B cells.In a EG7-OVA tumor-bearing murine model,APN-OVA-ZOL significantly inhibited the tumor growth and prolonged mice survival.This work provided a strong empirical foundation indicating that zoledronate-loaded aluminum salt nanovaccines had a strong potency for cancer immunotherapy.
基金The authors acknowledge support from the Tsinghua University Spring Breeze Fund,Center for Life Sciences,and Institute for Immunology,Tsinghua University,and grants from the Ministry of Science and Technology of China(2021YFC2300500 and 2021YFC2302403)National Natural Science Foundation of China(32141004,81825010,81730043,and 81621002).
文摘All organisms living in complex environments have evolved effective mechanisms of dynamic responses to extracellular stimuli.The immune system activates when damaged or injured cells release damage‐associated molecular patterns(DAMPs).In addition to well‐characterized DAMPs such as high‐mobility group box 1 and adenosine triphosphate,studies on new classes of DAMPs have emerged.Here,we review recent reports of a new class of isoprenoid‐derived DAMPs,including farnesyl pyrophosphate and geranylgeranyl pyrophosphate,both of which are pivotal metabolic inter-mediates of the mevalonate pathway.We also explore the roles of old and new DAMPs in autoimmune diseases that result from dysregulated inflammation.The findings highlight that understanding the functional mechanisms of DAMPs is important to enrich the DAMP family and decipher their immunoregulatory mechanisms to provide new therapeutics for the prevention and treatment of autoimmune diseases.
基金supported by National Natural Science Foundation of China to H.Z.and L.Z.(Grant No.81770184 and 81800174).
文摘Leukemia is a common hematological malignancy with overall poor prognosis.Novel therapies are needed to improve the outcome of leukemia patients.Cholesterol metabolism reprogramming is a featured alteration in leukemia.Many metabolic-related genes and metabolites are essential to the progress and drug resistance of leukemia.Exploring potential therapeutical targets related to cholesterol homeostasis is a promising area.This review summarized the functions of cholesterol and its derived intermediate metabolites,and also discussed potential agents targeting this metabolic vulnerability in leukemia.
基金This study was supported by Cancer Center Amsterdam grants 07/36 and 2012-1-08.
文摘Aim: This study aimed to decipher the molecular mechanism underlying the synergistic effect of inhibitors of the mevalonate-cholesterol pathway (i.e., statins) and aminopeptidase inhibitors (APis) on APi-sensitive and -resistant acute myeloid leukemia (AML) cells.Methods: U937 cells and their sublines with low and high levels of acquired resistance to (6S)-[(R)-2-((S)-Hydroxy-hydroxycarbamoyl-methoxy-methyl)-4-methyl-pentanoylamino]-3,3 dimethyl-butyric acid cyclopentyl ester (CHR2863), an APi prodrug, served as main AML cell line models. Drug combination effects were assessed with CHR2863 and in vitro non-toxic concentrations of various statins upon cell growth inhibition, cell cycle effects, and apoptosis induction. Mechanistic studies involved analysis of Rheb prenylation required for mTOR activation.Results: A strong synergy of CHR2863 with the statins simvastatin, fluvastatin, lovastatin, and pravastatin was demonstrated in U937 cells and two CHR2863-resistant sublines. This potent synergy between simvastatin and CHR2863 was also observed with a series of other human AML cell lines (e.g., THP1, MV4-11, and KG1), but not with acute lymphocytic leukemia or multiple solid tumor cell lines. This synergistic activity was: (i) specific for APis (e.g., CHR2863 and Bestatin), rather than for other cytotoxic agents;and (ii) corroborated by enhanced induction of apoptosis and cell cycle arrest which increased the sub-G1 fraction. Consistently, statin potentiation of CHR2863 activity was abrogated by co-administration of mevalonate and/or farnesyl pyrophosphate, suggesting the involvement of protein prenylation;this was experimentally confirmed by impaired Rheb prenylation by simvastatin.Conclusion: These novel findings suggest that the combined inhibitory effect of impaired Rheb prenylation and CHR2863-dependent mTOR inhibition instigates a potent synergistic inhibition of statins and APis on human AML cells.
基金supported by the Program for Scientific and Techno-logical Innovation Talents in Universities of Henan Provincial Education Department(25HASTIT057)the Henan Province Outstanding Youth Foundation(242300421097)National Natural Science Foundation of China(32371485 and 32111530179).
文摘Cycloastragenol is a bioactive,high-value triterpenoid derived from Astragalus membranaceus.Conventional plant-based extraction and chemical synthesis methods are expensive.To our knowledge,this is the first report on the de novo biosynthesis of cycloastragenol in yeast.The mevalonate pathway was reconstituted in yeast peroxisomes,and the engineered yeast produced 656.55 mg/L squalene.Further introduction of heterologous enzymes led the engineered yeast to produce 1.04 mg/L cycloastragenol,which demonstrated the yeast production of value-added medicinal molecules.
