Crosstalk between Kupffer cells(KCs)and hepatic stellate cells(HSCs)plays an important role in multiple liver disease conditions,including the formation of liver fibrosis in alcohol-associated liver disease(AALD).Ther...Crosstalk between Kupffer cells(KCs)and hepatic stellate cells(HSCs)plays an important role in multiple liver disease conditions,including the formation of liver fibrosis in alcohol-associated liver disease(AALD).Therapeutic targeting of the KC-HSC crosstalk is a prime target for therapeutic interventions.Herein,a novel modular nanosystem was designed and prepared through the self-assembly utilizing boric acid and catechol interactions to prepare polymers modified with a CXCR4-inhibiting moieties.The polymers were used to encapsulate anti-miR-155 and to block the undesirable crosstalk between HSCs and KCs by downregulating miR-155 expression in KCs with the parallel inhibition of CXCR4 signaling in activated HSCs.The combined inhibition of miR-155 and CXCR4 at two different liver cell types achieved improved antifibrosis effects in a mouse model of AALD fibrosis.Our finding highlights the key role that blocking the undesirable crosstalk between HSCs and KCs plays in reversing AALD fibrosis as well as demonstrates a proof-of-concept approach for designing and constructing multifunctional delivery nanosystems using orthogonal functional modules based on the understanding of disease mechanisms.展开更多
Tetracycline(TC)residues from anthropogenic activities undesirably present in nature as an emerging sustainability challenge and thereby require innovations in remediation technologies.Herein,as inspired by the microc...Tetracycline(TC)residues from anthropogenic activities undesirably present in nature as an emerging sustainability challenge and thereby require innovations in remediation technologies.Herein,as inspired by the microcompartment structure in living organisms,we adopt a synthetic biology approach to engineer the FerTiG,a modular enzyme assembly,to robustly scavenge TC residues with improved performance.The FerTiG consists of three functional modules,namely,a TC degradation module(Tet(X4)),a cofactor recycling module glucose dehydrogenase(GDH),and a protection module(ferritin),to organize diverse catalytic processes simultaneously as a biological circuit.The incorporation of GDH suitably fuels the FerTiG-dependent TC degradation by regenerating expensive nicotinamide adenine dinucleotide phosphate(NADPH)cofactor with glucose.The ferritin shields the catalytic core of FerTiG to resiliently decompose TC under unfavorable conditions.Due to collaboration among functional modules,FerTiG strongly catalyzes the residual TC removal from multiple environmental matrices.The degradation pathways and environmental/biological safety of FerTiG are then elaborated,indicating the promising readiness for the application of FerTiG.In summary,this work presents a synthetic biology-based strategy to spontaneously impose residual antibiotic biodegradation for better sustainability management.The FerTiG is engineered as a proof-of-principle for TC removal;however,this'microcompartment-mimick ing'concept is of great interest in mitigating other sustainability challenges where modular catalytic machinery is applied.展开更多
The introduction of living cells to manufacturing process has enabled the engineering of complex biological tissues in vitro.The recent advances in biofabrication with extremely high resolution(e.g.at single cell leve...The introduction of living cells to manufacturing process has enabled the engineering of complex biological tissues in vitro.The recent advances in biofabrication with extremely high resolution(e.g.at single cell level)have greatly enhanced this capacity and opened new avenues for tissue engineering.In this review,we comprehensively overview the current biofabrication strategies with single-cell resolution and categorize them based on the dimension of the single-cell building blocks,i.e.zero-dimensional single-cell droplets,one-dimensional single-cell filaments and two-dimensional single-cell sheets.We provide an informative introduction to the most recent advances in these approaches(e.g.cell trapping,bioprinting,electrospinning,microfluidics and cell sheets)and further illustrated how they can be used in in vitro tissue modelling and regenerative medicine.We highlight the significance of single-cell-level biofabrication and discuss the challenges and opportunities in the field.展开更多
Astaxanthin is a natural red carotenoid,commonly used as an additive in the pharmaceutical industry and as a nutritional supplement owing to its notable antioxidant benefits.However,a complex biosynthetic pathway pose...Astaxanthin is a natural red carotenoid,commonly used as an additive in the pharmaceutical industry and as a nutritional supplement owing to its notable antioxidant benefits.However,a complex biosynthetic pathway poses a challenge to de novo biosynthesis of astaxanthin.Here,Yarrowia lipolytica was engineered through multiple strategies for high level production of astaxanthin using a cheap mineral medium.For the production ofβ-carotene,a platform strain was constructed in which 411.7 mg/L ofβ-carotene was produced at a shake-flask level.Integration of algalβ-carotene ketolase andβ-carotene hydroxylase led to the production of 12.3 mg/L of astaxanthin.Furthermore,construction of HpBKT and HpCrtZ as a single enzyme complex along with the enhanced catalytic activity of the enzymes led to the accumulation of 41.0 mg/L of astaxanthin.Iterative gene integration into the genome and direction of the astaxanthin production pathway into sub-organelles substan-tially increased astaxanthin production(172.1 mg/L).Finally,restoration of the auxotrophic markers and me-dium optimization further improved astaxanthin production to 237.3 mg/L.The aforementioned approaches were employed in fed-batch fermentation to produce 2820 mg/L of astaxanthin(229-fold improvement regarding the starter strain),with an average productivity of 434 mg/L/d and a yield of 5.6 mg/g glucose,which is the highest reported productivity in Y.lipolytica.展开更多
基金This work was supported by a grant from the National Institute on Alcohol Abuse and Alcoholism(R01 AA027695)The Small Animal Ultrasound Core at UNMC is supported in part by funding from the Nebraska Center for Nanomedicine COBRE grant from the National Institute for General Medical Science(NIGMS)P30 GM127200.The UNMC Advanced Microscopy Core Facility receives partial support from the NIGMS INBRE-P20 GM103427 and COBRE-P30 GM106397 grants,as well as support from the National Cancer Institute(NCI)for The Fred&Pamela Buffett Cancer Center Support Grant-P30 CA036727the Nebraska Research Initiative.
文摘Crosstalk between Kupffer cells(KCs)and hepatic stellate cells(HSCs)plays an important role in multiple liver disease conditions,including the formation of liver fibrosis in alcohol-associated liver disease(AALD).Therapeutic targeting of the KC-HSC crosstalk is a prime target for therapeutic interventions.Herein,a novel modular nanosystem was designed and prepared through the self-assembly utilizing boric acid and catechol interactions to prepare polymers modified with a CXCR4-inhibiting moieties.The polymers were used to encapsulate anti-miR-155 and to block the undesirable crosstalk between HSCs and KCs by downregulating miR-155 expression in KCs with the parallel inhibition of CXCR4 signaling in activated HSCs.The combined inhibition of miR-155 and CXCR4 at two different liver cell types achieved improved antifibrosis effects in a mouse model of AALD fibrosis.Our finding highlights the key role that blocking the undesirable crosstalk between HSCs and KCs plays in reversing AALD fibrosis as well as demonstrates a proof-of-concept approach for designing and constructing multifunctional delivery nanosystems using orthogonal functional modules based on the understanding of disease mechanisms.
基金supported by the National Natural Science Foundation of China(32121004 and 32102720)the Guangzhou Science and Technology Plan Project(2024A04J6509)+3 种基金the National Key Research and Development Program of China(2023YFD1800100)the Local Innovative and Research Teams Project of Guangdong Pearl River Talents Program(2019BT02N054)the Double First-Class Discipline Promotion Project(2023B10564003)the 111 Project(D20008)。
文摘Tetracycline(TC)residues from anthropogenic activities undesirably present in nature as an emerging sustainability challenge and thereby require innovations in remediation technologies.Herein,as inspired by the microcompartment structure in living organisms,we adopt a synthetic biology approach to engineer the FerTiG,a modular enzyme assembly,to robustly scavenge TC residues with improved performance.The FerTiG consists of three functional modules,namely,a TC degradation module(Tet(X4)),a cofactor recycling module glucose dehydrogenase(GDH),and a protection module(ferritin),to organize diverse catalytic processes simultaneously as a biological circuit.The incorporation of GDH suitably fuels the FerTiG-dependent TC degradation by regenerating expensive nicotinamide adenine dinucleotide phosphate(NADPH)cofactor with glucose.The ferritin shields the catalytic core of FerTiG to resiliently decompose TC under unfavorable conditions.Due to collaboration among functional modules,FerTiG strongly catalyzes the residual TC removal from multiple environmental matrices.The degradation pathways and environmental/biological safety of FerTiG are then elaborated,indicating the promising readiness for the application of FerTiG.In summary,this work presents a synthetic biology-based strategy to spontaneously impose residual antibiotic biodegradation for better sustainability management.The FerTiG is engineered as a proof-of-principle for TC removal;however,this'microcompartment-mimick ing'concept is of great interest in mitigating other sustainability challenges where modular catalytic machinery is applied.
基金support from the National Natural Science Foundation of China(No.52105306,32211530075)New Faculty Start-up Funding provided by Tsinghua University(012-53330200421).
文摘The introduction of living cells to manufacturing process has enabled the engineering of complex biological tissues in vitro.The recent advances in biofabrication with extremely high resolution(e.g.at single cell level)have greatly enhanced this capacity and opened new avenues for tissue engineering.In this review,we comprehensively overview the current biofabrication strategies with single-cell resolution and categorize them based on the dimension of the single-cell building blocks,i.e.zero-dimensional single-cell droplets,one-dimensional single-cell filaments and two-dimensional single-cell sheets.We provide an informative introduction to the most recent advances in these approaches(e.g.cell trapping,bioprinting,electrospinning,microfluidics and cell sheets)and further illustrated how they can be used in in vitro tissue modelling and regenerative medicine.We highlight the significance of single-cell-level biofabrication and discuss the challenges and opportunities in the field.
基金supported by the National Key Research and Devel-opment Program of China(2022YFC2106100)the Foundation for Innovative Research Groups of the National Natural Science Foundation of China(32021005).
文摘Astaxanthin is a natural red carotenoid,commonly used as an additive in the pharmaceutical industry and as a nutritional supplement owing to its notable antioxidant benefits.However,a complex biosynthetic pathway poses a challenge to de novo biosynthesis of astaxanthin.Here,Yarrowia lipolytica was engineered through multiple strategies for high level production of astaxanthin using a cheap mineral medium.For the production ofβ-carotene,a platform strain was constructed in which 411.7 mg/L ofβ-carotene was produced at a shake-flask level.Integration of algalβ-carotene ketolase andβ-carotene hydroxylase led to the production of 12.3 mg/L of astaxanthin.Furthermore,construction of HpBKT and HpCrtZ as a single enzyme complex along with the enhanced catalytic activity of the enzymes led to the accumulation of 41.0 mg/L of astaxanthin.Iterative gene integration into the genome and direction of the astaxanthin production pathway into sub-organelles substan-tially increased astaxanthin production(172.1 mg/L).Finally,restoration of the auxotrophic markers and me-dium optimization further improved astaxanthin production to 237.3 mg/L.The aforementioned approaches were employed in fed-batch fermentation to produce 2820 mg/L of astaxanthin(229-fold improvement regarding the starter strain),with an average productivity of 434 mg/L/d and a yield of 5.6 mg/g glucose,which is the highest reported productivity in Y.lipolytica.
