Fluorescence lifetime imaging(FLIM)is increasingly used to read out cellular autofluorescenceoriginating from the coenzyme NADH in the context of investigating cell metabolic state.Wepresent here an automated multiwel...Fluorescence lifetime imaging(FLIM)is increasingly used to read out cellular autofluorescenceoriginating from the coenzyme NADH in the context of investigating cell metabolic state.Wepresent here an automated multiwell plate reading FLIM microscope optimized for UV illumi-nation with the goal of extending high content fluorescence lifetime asays to readouts ofmetabolism,We demonstrate its application to automated cellular autofluorescence lifetime imaging and discuss the key practical issues associated with its implementation.In particular,weillustrate its capability to read out the NADH-lifetime response of cells to metabolic modulators,thereby illustrating the potential of the instrument for cytotoxicity studies,assays for drugdiscovery and stratified medicine.展开更多
The forkhead transcription factors FOXO and FOXM1 have pivotal roles in tumorigenesis and in mediating chemotherapy sensitivity and resistance.Recent research shows that the forkhead transcription factor FOXM1 is a di...The forkhead transcription factors FOXO and FOXM1 have pivotal roles in tumorigenesis and in mediating chemotherapy sensitivity and resistance.Recent research shows that the forkhead transcription factor FOXM1 is a direct transcriptional target repressed by the forkhead protein FOXO3a,a vital downstream effector of the PI3K-AKT-FOXO signaling pathway.Intriguingly,FOXM1 and FOXO3a also compete for binding to the same gene targets,which have a role in chemotherapeutic drug action and sensitivity.An understanding of the role and regulation of the FOXO-FOXM1 axis will impact directly on our knowledge of chemotherapeutic drug action and resistance in patients,and provide new insights into the design of novel therapeutic strategy and reliable biomarkers for prediction of drug sensitivity.展开更多
基金funding from the UK Biotechnology and Biological Sciences Research Council(BBSRCBB/H00713X/1)the UK Engineering and PhysicalSciences Research Council(EPSRC EP/IO2770X/1)the UK Technology Strategy Board Technology Award(CHBT/007/00030,EP/C54269X,inpartnership with Astra Zeneca,GE Healthcare,GSK,Kentech Instruments Ltd).
文摘Fluorescence lifetime imaging(FLIM)is increasingly used to read out cellular autofluorescenceoriginating from the coenzyme NADH in the context of investigating cell metabolic state.Wepresent here an automated multiwell plate reading FLIM microscope optimized for UV illumi-nation with the goal of extending high content fluorescence lifetime asays to readouts ofmetabolism,We demonstrate its application to automated cellular autofluorescence lifetime imaging and discuss the key practical issues associated with its implementation.In particular,weillustrate its capability to read out the NADH-lifetime response of cells to metabolic modulators,thereby illustrating the potential of the instrument for cytotoxicity studies,assays for drugdiscovery and stratified medicine.
基金sponsored by Breast Cancer Campaign,Cancer Research UK,Imperial College NHS Health Trust.
文摘The forkhead transcription factors FOXO and FOXM1 have pivotal roles in tumorigenesis and in mediating chemotherapy sensitivity and resistance.Recent research shows that the forkhead transcription factor FOXM1 is a direct transcriptional target repressed by the forkhead protein FOXO3a,a vital downstream effector of the PI3K-AKT-FOXO signaling pathway.Intriguingly,FOXM1 and FOXO3a also compete for binding to the same gene targets,which have a role in chemotherapeutic drug action and sensitivity.An understanding of the role and regulation of the FOXO-FOXM1 axis will impact directly on our knowledge of chemotherapeutic drug action and resistance in patients,and provide new insights into the design of novel therapeutic strategy and reliable biomarkers for prediction of drug sensitivity.
