This study presents a simple and robust three-dimensional human hepatic tissue model to emulate steatotic and fibrotic conditions and provide an in vitro model for drug testing and mechanistic studies.Using a photolit...This study presents a simple and robust three-dimensional human hepatic tissue model to emulate steatotic and fibrotic conditions and provide an in vitro model for drug testing and mechanistic studies.Using a photolithographic biofabrication method with a photomask featuring hexagonal units,liver cells,including a human hepatic cell line(HepG2-C3A)and a human hepatic stellate cell line(LX-2)were embedded in gelatin methacryloyl hydrogel.Hepatic steatosis was induced by supraphysiological concentration of free fatty acids;hepatic fibrosis was induced by transforming growth factor-β1.Induction of steatosis was confirmed by Oil Red O and BODIPY staining and was inhibited with toyocamycin and obeticholic acid.Induction of fibrosis was confirmed by immunostaining for collagen type I and alpha smooth muscle actin and inhibited by rapamycin and curcumin treatment.This model was further preliminarily validated using primary human hepatocytes in a similar setup.These constructs provide a viable,biologically relevant,and higher throughput model of hepatic steatosis and fibrosis and may facilitate the study of the mechanisms of disease and testing of liver-directed drugs.展开更多
The microbiome,particularly the compo-sition and diversity of microbial communi-ties within various tissues,influences cancer development,progression and therapy response across various malignancies.1-5 Understanding ...The microbiome,particularly the compo-sition and diversity of microbial communi-ties within various tissues,influences cancer development,progression and therapy response across various malignancies.1-5 Understanding specific microbial signatures associated with cancers,termed the onco-biome,is crucial for advancing diagnostic and therapeutic strategies.展开更多
Background and Aims:Alpha-1 antitrypsin deficiency(AATD)is a genetic disorder associated with liver disease,ranging from fibrosis to hepatocellular carcinoma.The disease remains asymptomatic until its final stages whe...Background and Aims:Alpha-1 antitrypsin deficiency(AATD)is a genetic disorder associated with liver disease,ranging from fibrosis to hepatocellular carcinoma.The disease remains asymptomatic until its final stages when liver transplantation is the only available therapy.Biomarkers offer an advantage for disease evaluation.The presence of microRNAs(miRNAs)in plasma extracellular vesicles(EVs)presents a noninvasive approach to assess the molecular signatures of the disease.In this study,we aimed to identify miRNA biomarkers to distinguish molecular signatures of the liver disease associated with AATD in AATD individuals.Methods:Using small RNA sequencing and qPCR,we examined plasma EV miRNAs in healthy controls(n=20)and AATD patients(n=17).We compared the EV miRNAs of AATD individuals with and without liver disease,developing an approach for detecting liver disease.A set of miRNAs identified in the AATD testing cohort was validated in a separate cohort of AATD patients(n=45).Results:We identified differential expression of 178 EV miRNAs in the plasma of the AATD testing cohort compared to controls.We categorized AATD individuals into those with and without liver disease,identifying 39 differentially expressed miRNAs.Six miRNAs were selected to test their ability to discriminate liver disease in AATD.These were validated for their specificity and sensitivity in an independent cohort of 45 AATD individuals.Our logistic model established composite scores with threeand four-miRNA combinations,achieving areas under the curve of 0.737 and 0.751,respectively,for predicting AATD liver disease.Conclusions:We introduce plasma EV-derived miRNAs as potential biomarkers for evaluating AATD liver disease.Plasma EV-associated miRNAs may represent a molecular signature of AATD liver disease and could serve as valuable tools for its detection and monitoring.展开更多
Notch signaling plays important roles in the regulation of immune cell functioning during the inflammatory response.Activation of the innate immune signaling receptor NLRP3 promotes inflammation in injured tissue.Howe...Notch signaling plays important roles in the regulation of immune cell functioning during the inflammatory response.Activation of the innate immune signaling receptor NLRP3 promotes inflammation in injured tissue.However,it remains unknown whether Jagged1(JAG1)-mediated myeloid Notch1 signaling regulates NLRP3 function in acute liver injury.Here,we report that myeloid Notch1 signaling regulates the NLRP3-driven inflammatory response in ischemia/reperfusion(IR)-induced liver injury.In a mouse model of liver IR injury,Notch1-proficient(Notch1^(FL/FL))mice receiving recombinant JAG1 showed a reduction in IR-induced liver injury and increased Notch intracellular domain(NICD)and heat shock transcription factor 1(HSF1)expression,whereas myeloidspecific Notch1 knockout(Notch1^(M-KO))aggravated hepatocellular damage even with concomitant JAG1 treatment.Compared to JAG1-treated Notch1^(FL/FL) controls,Notch1^(M-KO) mice showed diminished HSF1 and Snail activity but augmented NLRP3/caspase-1 activity in ischemic liver.The disruption of HSF1 reduced Snail activation and enhanced NLRP3 activation,while the adoptive transfer of HSF1-expressing macrophages to Notch1^(M-KO) mice augmented Snail activation and mitigated IR-triggered liver inflammation.Moreover,the knockdown of Snail in JAG1-treated Notch1^(FL/FL) livers worsened hepatocellular functioning,reduced TRX1 expression and increased TXNIP/NLRP3 expression.Ablation of myeloid Notch1 or Snail increased ASK1 activation and hepatocellular apoptosis,whereas the activation of Snail increased TRX1 expression and reduced TXNIP,NLRP3/caspase-1,and ROS production.Our findings demonstrated that JAG1-mediated myeloid Notch1 signaling promotes HSF1 and Snail activation,which in turn inhibits NLRP3 function and hepatocellular apoptosis leading to the alleviation of IR-induced liver injury.Hence,the Notch1/HSF1/Snail signaling axis represents a novel regulator of and a potential therapeutic target for liver inflammatory injury.展开更多
基金YSZ received funding from National Institutes of Health(K99CA201603,R00CA201603,R21EB025270,R21EB026175,R01EB028143,R03EB027984)National Science Foundation(1935105)+1 种基金Brigham Research Institute New England Anti-Vivisection Foundation,and American Fund for Alternatives to Animal Research(AFAAR).AZ received funding from National Institutes of Health(K08DK113244,R01MD012579)SD received funding from National Institutes of Health(R01MD012579-UT20664DS).
文摘This study presents a simple and robust three-dimensional human hepatic tissue model to emulate steatotic and fibrotic conditions and provide an in vitro model for drug testing and mechanistic studies.Using a photolithographic biofabrication method with a photomask featuring hexagonal units,liver cells,including a human hepatic cell line(HepG2-C3A)and a human hepatic stellate cell line(LX-2)were embedded in gelatin methacryloyl hydrogel.Hepatic steatosis was induced by supraphysiological concentration of free fatty acids;hepatic fibrosis was induced by transforming growth factor-β1.Induction of steatosis was confirmed by Oil Red O and BODIPY staining and was inhibited with toyocamycin and obeticholic acid.Induction of fibrosis was confirmed by immunostaining for collagen type I and alpha smooth muscle actin and inhibited by rapamycin and curcumin treatment.This model was further preliminarily validated using primary human hepatocytes in a similar setup.These constructs provide a viable,biologically relevant,and higher throughput model of hepatic steatosis and fibrosis and may facilitate the study of the mechanisms of disease and testing of liver-directed drugs.
基金supported by R01 HL148801-02S1.AmZ is supported by the Prevent Cancer Foundation Grant,and NIH grants R21 CA259921,R01 AI163483,R01 EB030134 and U01 CA265719supported in part by the Department of Veterans Affairs,Veterans Health Administration,Office of Research and Development(VA Merit BLR&D Award I01 BX005707 to AmZ)+4 种基金RL receives funding support from NIH grants U01 DK061734,U01 DK130190,R01 DK106419,R01 DK121378,R01 DK124318 and P01 HL147835,as well as the John C Martin Foundation(RP124)KC is supported by AGA Research Foundation(AGA Research Scholar Award AGA2022-13-05)KC and CG are supported by NIH P30 CA023100 and R01 CA270235.AlZ has received funding support related to this study from NIH UH2 TR002087UH3 TR002087.RK is supported by NIH U24 CA248454,Pioneer DP1 AT010885 and R01 CA241728All authors receive institutional support from NIH P30 DK120515,P30 DK063491,P30 CA014195,P50 AA011999 and UL1 TR001442.
文摘The microbiome,particularly the compo-sition and diversity of microbial communi-ties within various tissues,influences cancer development,progression and therapy response across various malignancies.1-5 Understanding specific microbial signatures associated with cancers,termed the onco-biome,is crucial for advancing diagnostic and therapeutic strategies.
基金supported by a grant from the Alpha One Foundation(AGR00019116)the recipient of a Research Career Scientist Award from the Department of Veterans Affairs(IK6BX004477).
文摘Background and Aims:Alpha-1 antitrypsin deficiency(AATD)is a genetic disorder associated with liver disease,ranging from fibrosis to hepatocellular carcinoma.The disease remains asymptomatic until its final stages when liver transplantation is the only available therapy.Biomarkers offer an advantage for disease evaluation.The presence of microRNAs(miRNAs)in plasma extracellular vesicles(EVs)presents a noninvasive approach to assess the molecular signatures of the disease.In this study,we aimed to identify miRNA biomarkers to distinguish molecular signatures of the liver disease associated with AATD in AATD individuals.Methods:Using small RNA sequencing and qPCR,we examined plasma EV miRNAs in healthy controls(n=20)and AATD patients(n=17).We compared the EV miRNAs of AATD individuals with and without liver disease,developing an approach for detecting liver disease.A set of miRNAs identified in the AATD testing cohort was validated in a separate cohort of AATD patients(n=45).Results:We identified differential expression of 178 EV miRNAs in the plasma of the AATD testing cohort compared to controls.We categorized AATD individuals into those with and without liver disease,identifying 39 differentially expressed miRNAs.Six miRNAs were selected to test their ability to discriminate liver disease in AATD.These were validated for their specificity and sensitivity in an independent cohort of 45 AATD individuals.Our logistic model established composite scores with threeand four-miRNA combinations,achieving areas under the curve of 0.737 and 0.751,respectively,for predicting AATD liver disease.Conclusions:We introduce plasma EV-derived miRNAs as potential biomarkers for evaluating AATD liver disease.Plasma EV-associated miRNAs may represent a molecular signature of AATD liver disease and could serve as valuable tools for its detection and monitoring.
基金supported by the NIH grants R01AI139552,R21AI146742,R21AI112722,R21AI115133(B.K.),P01AI120944,R01DK062357,R01DK102110,and R01DK107533(J.W.K.-W.)by the Dumont Research Foundation.
文摘Notch signaling plays important roles in the regulation of immune cell functioning during the inflammatory response.Activation of the innate immune signaling receptor NLRP3 promotes inflammation in injured tissue.However,it remains unknown whether Jagged1(JAG1)-mediated myeloid Notch1 signaling regulates NLRP3 function in acute liver injury.Here,we report that myeloid Notch1 signaling regulates the NLRP3-driven inflammatory response in ischemia/reperfusion(IR)-induced liver injury.In a mouse model of liver IR injury,Notch1-proficient(Notch1^(FL/FL))mice receiving recombinant JAG1 showed a reduction in IR-induced liver injury and increased Notch intracellular domain(NICD)and heat shock transcription factor 1(HSF1)expression,whereas myeloidspecific Notch1 knockout(Notch1^(M-KO))aggravated hepatocellular damage even with concomitant JAG1 treatment.Compared to JAG1-treated Notch1^(FL/FL) controls,Notch1^(M-KO) mice showed diminished HSF1 and Snail activity but augmented NLRP3/caspase-1 activity in ischemic liver.The disruption of HSF1 reduced Snail activation and enhanced NLRP3 activation,while the adoptive transfer of HSF1-expressing macrophages to Notch1^(M-KO) mice augmented Snail activation and mitigated IR-triggered liver inflammation.Moreover,the knockdown of Snail in JAG1-treated Notch1^(FL/FL) livers worsened hepatocellular functioning,reduced TRX1 expression and increased TXNIP/NLRP3 expression.Ablation of myeloid Notch1 or Snail increased ASK1 activation and hepatocellular apoptosis,whereas the activation of Snail increased TRX1 expression and reduced TXNIP,NLRP3/caspase-1,and ROS production.Our findings demonstrated that JAG1-mediated myeloid Notch1 signaling promotes HSF1 and Snail activation,which in turn inhibits NLRP3 function and hepatocellular apoptosis leading to the alleviation of IR-induced liver injury.Hence,the Notch1/HSF1/Snail signaling axis represents a novel regulator of and a potential therapeutic target for liver inflammatory injury.
