A meso–molecular muscle was prepared by capping the[c2]daisy chain based on a mono-functionalized copillar[5]arene with an imidazolium group in its axle.From its crystal structure,we observed that it was a cyclic dim...A meso–molecular muscle was prepared by capping the[c2]daisy chain based on a mono-functionalized copillar[5]arene with an imidazolium group in its axle.From its crystal structure,we observed that it was a cyclic dimer composed of two mirror image subcomponents,a pR-and a pS-copillar[5]arene.Their conformations were fixed by the doubly interlocked mechanical bond.By comparison of the^1H NMR and COSY spectra,we found that the length of this meso–molecular muscle could be controlled not only by the solvents,but also by the counter anions.展开更多
Tissue engineering provides a promising approach for regenerative medicine.The ideal engineered tissue should have the desired structure and functional properties suitable for uniform cell distribution and stable shap...Tissue engineering provides a promising approach for regenerative medicine.The ideal engineered tissue should have the desired structure and functional properties suitable for uniform cell distribution and stable shape fidelity in the full period of in vitro culture and in vivo implantation.However,due to insufficient cell infiltration and inadequate mechanical properties,engineered tissue made from porous scaffolds may have an inconsistent cellular composition and a poor shape retainability,which seriously hinders their further clinical application.In this study,silk fibroin was integrated with silk short fibers with a physical and chemical double-crosslinking network to fabricate fiber-reinforced silk fibroin super elastic absorbent sponges(Fr-SF-SEAs).The Fr-SF-SEAs exhibited the desirable synergistic properties of a honeycomb structure,hygroscopicity and elasticity,which allowed them to undergo an unconventional cyclic compression inoculation method to significantly promote cell diffusion and achieve a uniform cell distribution at a high-density.Furthermore,the regenerated cartilage of the Fr-SF-SEAs scaffold withstood a dynamic pressure environment after subcutaneous implantation and maintained its precise original structure,ultimately achieving human-scale ear-shaped cartilage regeneration.Importantly,the SF-SEAs prepara-tion showed valuable universality in combining chemicals with other bioactive materials or drugs with reactive groups to construct microenvironment bionic scaffolds.The established novel cell inoculation method is highly versatile and can be readily applied to various cells.Based on the design concept of dual-network Fr-SF-SEAs scaffolds,homogenous and mature cartilage was successfully regenerated with precise and complicated shapes,which hopefully provides a platform strategy for tissue engineering for various cartilage defect repairs.展开更多
Background:The efficacy of immune checkpoint blockade therapy in patients with hepatocellular carcinoma(HCC)remains poor.Although serine-and arginine-rich splicing factor(SRSF)family members play crucial roles in tumo...Background:The efficacy of immune checkpoint blockade therapy in patients with hepatocellular carcinoma(HCC)remains poor.Although serine-and arginine-rich splicing factor(SRSF)family members play crucial roles in tumors,their impact on tumor immunology remains unclear.This study aimed to elucidate the role of SRSF10 in HCC immunotherapy.Methods:To identify the key genes associated with immunotherapy resistance,we conducted single-nuclear RNA sequencing,multiplex immunofluorescence,and The Cancer Genome Atlas and Gene Expression Omnibus database analyses.We investigated the biological functions of SRSF10 in immune evasion using in vitro co-culture systems,flow cytometry,various tumor-bearing mouse models,and patient-derived organotypic tumor spheroids.Results:SRSF10 was upregulated in various tumors and associated with poor prognosis.Moreover,SRSF10 positively regulated lactate production,and SRSF10/glycolysis/histone H3 lysine 18 lactylation(H3K18la)formed a positive feedback loop in tumor cells.Increased lactate levels promoted M2 macrophage polarization,thereby inhibiting CD8^(+)T cell activity.Mechanistically,SRSF10 interacted with the 3′-untranslated region of MYB,enhancing MYB RNA stability,and subsequently upregulating key glycolysis-related enzymes including glucose transporter 1(GLUT1),hexokinase 1(HK1),lactate dehydrogenase A(LDHA),resulting in elevated intracellular and extracellular lactate levels.Lactate accumulation induced histone lactylation,which further upregulated SRSF10 expression.Additionally,lactate produced by tumors induced lactylation of the histone H3K18la site upon transport into macrophages,thereby activating transcription and enhancing pro-tumor macrophage activity.M2 macrophages,in turn,inhibited the enrichment of CD8^(+)T cells and the proportion of interferon-γ+CD8^(+)T cells in the tumor microenvironment(TME),thus creating an immunosuppressive TME.Clinically,SRSF10 could serve as a biomarker for assessing immunotherapy resistance in various solid tumors.Pharmacological targeting of SRSF10 with a selective inhibitor 1C8 enhanced the efficacy of programmed cell death 1(PD-1)monoclonal antibodies(mAbs)in both murine and human preclinical models.Conclusions:The SRSF10/MYB/glycolysis/lactate axis is critical for triggering immune evasion and anti-PD-1 resistance.Inhibiting SRSF10 by 1C8 may overcome anti-PD-1 tolerance in HCC.展开更多
Cartilage tissue engineering is a promising strategy for repairing cartilage defects.However,achieving satisfactory cartilage regeneration in vitro and maintaining its stability in vivo remains a challenge.The key to ...Cartilage tissue engineering is a promising strategy for repairing cartilage defects.However,achieving satisfactory cartilage regeneration in vitro and maintaining its stability in vivo remains a challenge.The key to achieving this goal is establishing an efficient cartilage regeneration culture system to retain sufficient active cells with physiological functions,generate abundant cartilage extracellular matrix(ECM)and maintain a low level of cartilage ECM degradation.The current chondrogenic medium(CM)can effectively promote cartilage ECM production;however,it has a negative effect on cell proliferation.Meanwhile,the specific c-Jun N-terminal kinase pathway inhibitor SP600125 promotes chondrocyte proliferation but inhibits ECM synthesis.Here,we aimed to construct a three-dimensional cartilage regeneration model using a polyglycolic acid/polylactic acid scaffold in combination with chondrocytes to investigate the effect of different culture modes with CM and SP600125 on in vitro cartilage regeneration and their long-term outcomes in vivo systematically.Our results demonstrate that the long-term combination of CM and SP600125 made up for each other and maximized their respective advantages to obtain optimal cartilage regeneration in vitro.Moreover,the long-term combination achieved stable cartilage regeneration after implantation in vivo with a relatively low initial cell-seeding concentration.Therefore,the long-term combination of CM and SP600125 enhanced in vitro and in vivo cartilage regeneration stability with fewer initial seeding cells and thus optimized the cartilage regeneration culture system.展开更多
Despite the notable efficacy of anti-PD1 therapy in the management of hepatocellular carcinoma(HCC)patients,resistance in most individuals necessitates additional investigation.For this study,we collected tumor tissue...Despite the notable efficacy of anti-PD1 therapy in the management of hepatocellular carcinoma(HCC)patients,resistance in most individuals necessitates additional investigation.For this study,we collected tumor tissues from nine HCC patients receiving anti-PD1 monotherapy and conducted RNA sequencing.These findings revealed significant upregulation of GSDME,which is predominantly expressed by tumor-associated macrophages(TAMs),in anti-PD1-resistant patients.Furthermore,patients with elevated levels of GSDME+macrophages in HCC tissues presented a poorer prognosis.The analysis of single-cell sequencing data and flow cytometry revealed that the suppression of GSDME expression in nontumor cells resulted in a decrease in the proportion of M2-like macrophages within the tumor microenvironment(TIME)of HCC while concurrently augmenting the cytotoxicity of CD8+T cells.The non-N-terminal fragment of GSDME within macrophages combines with PDPK1,thereby activating the PI3K-AKT pathway and facilitating M2-like polarization.The small-molecule Eliprodil inhibited the increase in PDPK1 phosphorylation mediated by GSDME site 1.The combination of Eliprodil and anti-PD1 was effective in the treatment of both spontaneous HCC in c-Myc+/+;Alb-Cre+/+mice and in a hydrodynamic tail vein injection model,which provides a promising strategy for novel combined immunotherapy.展开更多
基金the Natural Science Foundation of Zhejiang Province(No.LY22B040001)the Science&Technology Innovation Program of Zhejiang Province(No.2018R52051)the National Natural Science Foundation of China(No.22035006)for financial support。
文摘A meso–molecular muscle was prepared by capping the[c2]daisy chain based on a mono-functionalized copillar[5]arene with an imidazolium group in its axle.From its crystal structure,we observed that it was a cyclic dimer composed of two mirror image subcomponents,a pR-and a pS-copillar[5]arene.Their conformations were fixed by the doubly interlocked mechanical bond.By comparison of the^1H NMR and COSY spectra,we found that the length of this meso–molecular muscle could be controlled not only by the solvents,but also by the counter anions.
基金support from the National Key Research and Development Program of China(2018YFC1105800,2017YFC1103900)the National Natural Science Foundation of China(82102211,81871502)+5 种基金the Shanghai Municipal Key Clinical Specialty(shslczdzk06601)the Shanghai Collaborative Innovation Program on Regenerative Medicine and Stem Cell Research(2019CXJQ01)the Key Research and Development Program of Henan Province(No.221111310100)the Major Science and Technology Projects of Xinxiang City(No.21ZD006)the Clinical Research Plan of SHDC(No.SHDC2020CR2045B)the Start-up Funds of Talent Construction and Scientific Research in Shanghai 9th People's Hospital(2021rcyj-ld).
文摘Tissue engineering provides a promising approach for regenerative medicine.The ideal engineered tissue should have the desired structure and functional properties suitable for uniform cell distribution and stable shape fidelity in the full period of in vitro culture and in vivo implantation.However,due to insufficient cell infiltration and inadequate mechanical properties,engineered tissue made from porous scaffolds may have an inconsistent cellular composition and a poor shape retainability,which seriously hinders their further clinical application.In this study,silk fibroin was integrated with silk short fibers with a physical and chemical double-crosslinking network to fabricate fiber-reinforced silk fibroin super elastic absorbent sponges(Fr-SF-SEAs).The Fr-SF-SEAs exhibited the desirable synergistic properties of a honeycomb structure,hygroscopicity and elasticity,which allowed them to undergo an unconventional cyclic compression inoculation method to significantly promote cell diffusion and achieve a uniform cell distribution at a high-density.Furthermore,the regenerated cartilage of the Fr-SF-SEAs scaffold withstood a dynamic pressure environment after subcutaneous implantation and maintained its precise original structure,ultimately achieving human-scale ear-shaped cartilage regeneration.Importantly,the SF-SEAs prepara-tion showed valuable universality in combining chemicals with other bioactive materials or drugs with reactive groups to construct microenvironment bionic scaffolds.The established novel cell inoculation method is highly versatile and can be readily applied to various cells.Based on the design concept of dual-network Fr-SF-SEAs scaffolds,homogenous and mature cartilage was successfully regenerated with precise and complicated shapes,which hopefully provides a platform strategy for tissue engineering for various cartilage defect repairs.
基金supported by the National Natural Science Foundation of China(No.82372946 and No.82072670)the 12 Leading Project of the Science and Technology Committee of Shanghai Municipality(No.21Y21900100)the Project of Shanghai Municipal Health Commission(No.202140269).
文摘Background:The efficacy of immune checkpoint blockade therapy in patients with hepatocellular carcinoma(HCC)remains poor.Although serine-and arginine-rich splicing factor(SRSF)family members play crucial roles in tumors,their impact on tumor immunology remains unclear.This study aimed to elucidate the role of SRSF10 in HCC immunotherapy.Methods:To identify the key genes associated with immunotherapy resistance,we conducted single-nuclear RNA sequencing,multiplex immunofluorescence,and The Cancer Genome Atlas and Gene Expression Omnibus database analyses.We investigated the biological functions of SRSF10 in immune evasion using in vitro co-culture systems,flow cytometry,various tumor-bearing mouse models,and patient-derived organotypic tumor spheroids.Results:SRSF10 was upregulated in various tumors and associated with poor prognosis.Moreover,SRSF10 positively regulated lactate production,and SRSF10/glycolysis/histone H3 lysine 18 lactylation(H3K18la)formed a positive feedback loop in tumor cells.Increased lactate levels promoted M2 macrophage polarization,thereby inhibiting CD8^(+)T cell activity.Mechanistically,SRSF10 interacted with the 3′-untranslated region of MYB,enhancing MYB RNA stability,and subsequently upregulating key glycolysis-related enzymes including glucose transporter 1(GLUT1),hexokinase 1(HK1),lactate dehydrogenase A(LDHA),resulting in elevated intracellular and extracellular lactate levels.Lactate accumulation induced histone lactylation,which further upregulated SRSF10 expression.Additionally,lactate produced by tumors induced lactylation of the histone H3K18la site upon transport into macrophages,thereby activating transcription and enhancing pro-tumor macrophage activity.M2 macrophages,in turn,inhibited the enrichment of CD8^(+)T cells and the proportion of interferon-γ+CD8^(+)T cells in the tumor microenvironment(TME),thus creating an immunosuppressive TME.Clinically,SRSF10 could serve as a biomarker for assessing immunotherapy resistance in various solid tumors.Pharmacological targeting of SRSF10 with a selective inhibitor 1C8 enhanced the efficacy of programmed cell death 1(PD-1)monoclonal antibodies(mAbs)in both murine and human preclinical models.Conclusions:The SRSF10/MYB/glycolysis/lactate axis is critical for triggering immune evasion and anti-PD-1 resistance.Inhibiting SRSF10 by 1C8 may overcome anti-PD-1 tolerance in HCC.
基金supported by the National Key Research and Development Program of China(2017YFC1103900 and 2018YFC1105800)the National Natural Science Foundation of China(81871502 and 81701843)+3 种基金the Program of Shanghai Academic/Technology Research Leader(19XD1431100)the Shanghai Collaborative Innovation Program on Regenerative Medicine and Stem Cell Research(2019CXJQ01)the Clinical Research Plan of SHDC(No.SHDC2020CR2045B),Shanghai Municipal Key Clinical Specialty(shslczdzk06601)Biomaterials and Regenerative Medicine Institute Cooperative Research Project,Shanghai Jiao Tong University School of Medicine(2022LHA07).
文摘Cartilage tissue engineering is a promising strategy for repairing cartilage defects.However,achieving satisfactory cartilage regeneration in vitro and maintaining its stability in vivo remains a challenge.The key to achieving this goal is establishing an efficient cartilage regeneration culture system to retain sufficient active cells with physiological functions,generate abundant cartilage extracellular matrix(ECM)and maintain a low level of cartilage ECM degradation.The current chondrogenic medium(CM)can effectively promote cartilage ECM production;however,it has a negative effect on cell proliferation.Meanwhile,the specific c-Jun N-terminal kinase pathway inhibitor SP600125 promotes chondrocyte proliferation but inhibits ECM synthesis.Here,we aimed to construct a three-dimensional cartilage regeneration model using a polyglycolic acid/polylactic acid scaffold in combination with chondrocytes to investigate the effect of different culture modes with CM and SP600125 on in vitro cartilage regeneration and their long-term outcomes in vivo systematically.Our results demonstrate that the long-term combination of CM and SP600125 made up for each other and maximized their respective advantages to obtain optimal cartilage regeneration in vitro.Moreover,the long-term combination achieved stable cartilage regeneration after implantation in vivo with a relatively low initial cell-seeding concentration.Therefore,the long-term combination of CM and SP600125 enhanced in vitro and in vivo cartilage regeneration stability with fewer initial seeding cells and thus optimized the cartilage regeneration culture system.
基金supported by the National Natural Science Foundation of China(No.82372946,No.82072670,and No.81871916)the Leading Project of the Science and Technology Committee of Shanghai Municipality(No.21Y21900100)the Project of Shanghai Municipal Health Commission(No.202140269).
文摘Despite the notable efficacy of anti-PD1 therapy in the management of hepatocellular carcinoma(HCC)patients,resistance in most individuals necessitates additional investigation.For this study,we collected tumor tissues from nine HCC patients receiving anti-PD1 monotherapy and conducted RNA sequencing.These findings revealed significant upregulation of GSDME,which is predominantly expressed by tumor-associated macrophages(TAMs),in anti-PD1-resistant patients.Furthermore,patients with elevated levels of GSDME+macrophages in HCC tissues presented a poorer prognosis.The analysis of single-cell sequencing data and flow cytometry revealed that the suppression of GSDME expression in nontumor cells resulted in a decrease in the proportion of M2-like macrophages within the tumor microenvironment(TIME)of HCC while concurrently augmenting the cytotoxicity of CD8+T cells.The non-N-terminal fragment of GSDME within macrophages combines with PDPK1,thereby activating the PI3K-AKT pathway and facilitating M2-like polarization.The small-molecule Eliprodil inhibited the increase in PDPK1 phosphorylation mediated by GSDME site 1.The combination of Eliprodil and anti-PD1 was effective in the treatment of both spontaneous HCC in c-Myc+/+;Alb-Cre+/+mice and in a hydrodynamic tail vein injection model,which provides a promising strategy for novel combined immunotherapy.
