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imaging redox STATE HETEROGENEITY WITHIN INDIVIDUAL EMBRYONIC STEM CELL COLONIES

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Journal of Innovative Optical Health Sciences 2011年第3期4卷 279-288页

作者： HE N.XU RUSSELL C.ADDIS DAVIDA F.GOINGS SHOKO NIOKA britton chance JOHN D.GEARHART LIN Z.LI Molecular imaging laboratory Department of Radiology University of PennsylvaniaSchool of Medicine PhiladelphiaPA 19104USA Institute for Regenerative Medicine and department of Cell and Developmental Biology University of Pennsylvania School of Medicine PhiladelphiaPA 19104USA britton chance laboratory of redox imaging Johnson Research Foundation department of Biochemistry and Biophysics University of Pennsylvania School of Medicine PhiladelphiaPA 19104USA Institute of Translational Medicine and Therapeutics University of Pennsylvania PhiladelphiaPA 19104USA

redox state mediates embryonic stem cell(ESC)differentiation and thus offers an important complementary approach to understanding the pluripotency of stem *** redox ratio(NADH/(Fp t NADH)),where NADH is the reduced form of nicotinamide adenine dinucleotide and Fp is the oxidizedflavoproteins,has been established as a sensitive indicator of mitochondrial redox *** this paper,we report our redox imaging data on the mitochondrial redox state of mouse ESC(mESC)colonies and the implications *** low-temperature NADH/Fp redox scanner was employed to image mESC colonies grown on a feeder layer of gamma-irradiated mouse embryonicfibroblasts(MEFs)on glass cover *** result showed significant heterogeneity in the mitochondrial redox state within individual mESC colonies(size:~200-440μm),exhibiting a core with a more reduced state than the *** more reduced state positively correlates with the expression pattern of Oct4,a well-established marker of *** observation is thefirst to show the heterogeneity in the mitochondrial redox state within a mESC colony,suggesting that mitochondrial redox state should be further investigated as a potential new biomarker for the stemness of embryonic stem cells.

关键词： redox imaging pluripotency NADH flavoproteins redox ratio

INTRODUCTION SPECIAL ISSUE IN MEMORY OF PROFESSOR britton chance

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Journal of Innovative Optical Health Sciences 2011年第3期4卷 I0001-I0003页

作者： Lin Z.Li Shoko Nioka Molecular imaging Lab Department of Radiology britton chance Lab of redox imaging Johnson Foundation department of Biochemistry and Biophysics University of Pennsylvania

britton chance:One of the Most Outstanding Scientists in the World On June 3rd and 4th,2011,over 300 scientists and professionals from all over the world gathered at the Translational Research Center of the University of Pennsylvania to attend a memorial symposium on\britton chance:His Life,Times and Legacy"and a\Molecular Spectroscopy/imaging Workshop:from bench top to bedside"dedicated to britton chance as well.

关键词： Times chance symposium

redox imaging OF THE p53-DEPENDENT MITOCHONDRIAL redox STATE IN COLON CANCER EX VIVO

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Journal of Innovative Optical Health Sciences 2013年第3期6卷 1-10页

作者： HE N.XU MIN FENG LILY MOON NATHAN DOLLOFF WAFIKEL-DEIRY LIN Z.LI department of radiology University of Pennsylvania PhiladelphiaPAUSA britton chance laboratory of redox imaging Johnson Research Foundation department of Biochemistry and Biophysics Perelman School of Medicine University of Pennsylvania PhiladelphiaPAUSA department of Medicine Penn State Hershey Medical Center and Penn State College of MedicineHersheyPAUSA Hematology/Oncology Penn State Hershey Medical Center HersheyPAUSA Abramson Cancer Center University of Pennsylvania PhiladelphiaPAUSA Institute of Translational Medicine and Therapeutics University of Pennsylvania PhiladelphiaPAUSA

The mitochondrial redox state and its heterogeneity of colon cancer at tissue level have not been previously *** has how p53 regulates mitochondial respi ration been measured at(deep)tissue level,presumably due to the unavailability of the technology that has sufficient spatial resolution and tissue penetration *** prior work demonstrated that the mito-chondrial redox state and its intnatumor heterogeneity is associated with cancer aggressiveness in human melanoma and breast cancer in mouse models,with the more metastatic tumons exhi-bit ing localized negions of more oxidized redox *** the chance redox scanner with an in-plane spatial resolution of 200 pm,we imaged the mitochondrial redox state of the wild-type p53 colon tumors(HCT116 p53 ut)and the p5-deleted colon tumors(HCT116 p53-/-)by cllcting the fuorescence si gnals of nicotinamide adenine dimucleotide(NA DH)and oxidized flavoproteins [Fp,including favin adenine dinucleotide(FAD)]from the mouse xenogmafts snap frozen at low *** results show that:(1)both tumor lines have significant degree of intratumor heterogeneity of the redox state,typically exhibiting a distinct bi modal distribution that either correlates with the spatial core-rim pattern or the“hot/oold”oxida tion-roduction patches;.(2)the p531-group is significantly more beterogencous in the mitochondrial redox state and has a more oxidized turmor core compared to the p53 wt group when the tunor sizes of the two groups are matched;(3)the tumor size dependence of the redox indices(such as Fp and Fp redox ratio)is significant in the p531-group with the larger ones being more oxidized and more hetero-geneous in their redox state,particularly more oxidized in the tumor central regions;(4)the H&E staining images of tumor soctions grossly correlate with the redox *** present work is the first to reveal at the submillimeter scale the intratumor heterogeneity pattem of the mitochon-drial redox state in colon cancer and

关键词： NADH favoprotein intratumor heterogeneity HCT116 p53 null necrotic

CHOP THER APY INDUCED MITOCHONDRIAL redox STATE ALTERATION IN NON-HODGKIN'S LYMPHOMA XENOGRAFTS

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Journal of Innovative Optical Health Sciences 2013年第2期6卷 29-35页

作者： H.N.XU H.ZHAO T.A.MIR S.C.LEE M.FENG R.CHOE J.D.GLICKSON L.Z.LI department of radiology University of Pennsylvania PhiladelphiaPAUSA britton chance laboratory of redox imaging Johnson Research Foundation department of Biochemistry and Biophysics University of Pennsylvania PhiladelphiaPAUSA Biostatistics and Data Management Core The Children's Hospital of Philadelphia(CHOP) PhiladelphiaPAUSA department of Biomedical Engineering University of Rochester RochesterNYUSA Abramson Cancer Center University of Pennsylvania PhiladelphiaPAUSA Institute of Translational Medicine and Therapeutics University of Pennsylvania PhiladelphiaPAUSA

We are interested in investigating whether cancer therapy may alter the mitochondrial redox state in cancer cells to inhibit their growth and *** redox state can be imaged by the redox scanner that collects the fuorescence signals from both the oxidized-fAavoproteins(Fp)and the reduced form of nicotinamide adenine dinucleotide(NADH)in snap frozen tissues and has been previously employed to study tumor aggressiveness and treatment ***,with the redox scanner we investigated the effects of chemotherapy on mouse xenografts of a human diffuse large *** lymphoma cell line(DLCL2).The mice were treated with CHOP therapy,i.e,cyclophosphamide(C)+hydroxydoxorubicin(H)+Oncovin(O)+prednisone(P)with CHO administration on day 1 and prednisone administration on days *** Fp content of the treated group was significantly decreased(p=0.033)on day 5,and the mitochondrial redox state of the treated group was slightly more reduced than that of the control group(p=0.048).The decrease of the Fp heterogeneity(measured by the mean st andard deviation)had a border-line statistical significance(p=0.071).The result suggests that the mitochondrial metabolism of lymphoma cells was slightly suppressed and the lymphomas became less aggressive after the CHOP therapy.

关键词： NA DH flavoprotein DLCL2 therapeutic effect tumor metabolism

Optical redox imaging of ANT1-de-cient muscles

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Journal of Innovative Optical Health Sciences 2024年第1期17卷 135-140页

作者： He N.Xu Ryan M.Morrow Min Feng Huaqing Zhao Douglas Wallace Lin Z.Li britton chance laboratory of redox imaging department of radiology Perelman School of Medicine University of Pennsylvania PhiladelphiaPA 19104USA Center for Mitochondrial and Epigenomic Medicine Children's Hospital of Philadelphia Research Institute Philadelphia PA 19104USA Center for Biostatistics and Epidemiology Lewis Katz School of Medicine at Temple University Philadelphia PA 19140USA

Adenine nucleotide translocator(ANT)is a mitochondrial protein involved in the exchange of ADP and ATP across the mitochondrial inner *** plays a crucial role in cellular energy metabolism by facilitating the transport of ATP synthesized within the mitochondria to the *** isoform ANT1 predominately expresses in cardiac and skeletal *** or dysregulation in ANT1 have been implicated in various mitochondrial disorders and neuromuscular *** aimed to examine whether ANT1 deletion may affect mitochondrial redox state in our established ANT1-de-cient *** and quadriceps resected from age-matched wild type(WT)and ANT1-de-cient mice were snap-frozen in liquid *** chance redox scanner was utilized to perform 3D optical redox *** sample underwent scanning across 3–5 *** averaging analysis showed no signi-cant differences in the redox indices(NADH,flavin adenine dinucleotide containing-flavoproteins Fp,and the redox ratio Fp/(NADH+Fp)between WT and ANT1-de-cient ***,quadriceps had higher Fp than hearts in both groups(p¼0:0004 and 0.01,respectively).Furthermore,the quadriceps were also more oxidized(a higher redox ratio)than hearts in WT group(p¼0:004).NADH levels were similar in all *** data suggest that under non-stressful physical condition,the ANT1-de-cient muscle cells were in the same mitochondrial state as WT ones and that the signi-cant difference in the mitochondrial redox state between quadriceps and hearts found in WT might be diminished in ANT1-de-cient *** imaging of muscles under physical stress can be conducted in future.

关键词： ANT1 redox ratio flavoproteins

Fluorescent and quantitative mitochondrial redox imaging of tumor targeted by Octa-RGD probe

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Journal of Innovative Optical Health Sciences 2016年第4期9卷 55-65页

作者： Shuang Sha Fei Yang Anle Wang Honglin Jin Zhihong Zhang Qiaoya Lin britton chance Center for Biomedical Photonics Wuhan National laboratory for Optoelectronics–Huazhong University of Science and Technology Wuhan 430074 P.R.China MoE Key laboratory for Biomedical Photonics department of Biomedical Engineering Huazhong University of Science and Technology Wuhan 430074 P.R.China

Integrins,over-expressed in a broad range of cancer diseauses,are widely utilized as a tumor *** investigation also plays important roles in tumor ***-oping simple integrin-targetting probe and monitoring tumor metabolism will give opportunities to find ways for cancer treatment,however,the investigation of tumor metabolism with integrin receptor based probes has been rarely reported so ***,we developed an octavalent fuo-rescent probe *** by convenient genetic method,based on one tetrameric far-red fluo-rescent protein(fRFP)linked with RGD pept *** validated its inter gin targeting by confocal imaging in *** we screened a variety of tumor cells,and differentiated their binding affinity based on the fuorescence of the probe via flow *** these cells,CNE-2 cells had the highest uptake of the probe,while B16 cells had the lowest,corresponding with their intergin expression ***,the fuorescent and metabolic imaging was performed in HT1080(intergin postive)tumor,where nicotinamide adenine dinudeotide hydrogen(NADH),flavo-protein(Fp)and fRFP fuorescent signals were *** tumor from mice intravenously injected with Octa RGD probe displayed obviously higher NADH redox ratio NADH/(Fp+NADH)and fRFP signal,than those with fRFP *** suggested that integrin targeting may have influence on the target cell metabolism,and further demonstrated *** probe facilitated its uptake in the targeted tumor in *** paper developed a useful probe,which can bind integrins speci-cally and e±ciently in tumor cells,and together with tumor metabolic information,it may provide new insight for RGD targeting-based cancer therapeutics.

关键词： Integrin redox metabolism fRFP NADH Fp.

KillerRed protein based in vivo photodynamic therapy and corresponding tumor metabolic imaging

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Journal of Innovative Optical Health Sciences 2016年第1期9卷 21-30页

作者： Qiaoya Lin Shuang Sha Fei Yang Honglin Jin Zhihong Zhang britton chance Center for Biomedical Photonics Wuhan National laboratory for Optoelectronics Huazhong University of Science and Technology Wuhan 430074 P.R.China MoE Key laboratory for Biomedical Photonics department of Biomedical Engineering Huazhong University of Science and Technology Wuhan 430074 P.R.China

Photodynamic therapy(PDT)gains wide attention as a useful therapeutic method for *** is mediated by the oxygen and photosensitizer under the specific light irradiation to produce the reactive oxygen species(ROS),which induce cellular toxicity and regulate the redox potential in tumor ***,genetic photosensitizers of low toxicity and easy production are required to be ***,a unique red fluorescent protein exhibiting excellent phototoxic properties,has the potential to act as a photosensitizer in the application of tumor ***,the course of tumor redox metabolism during this treatment was rarely investigated so *** here,we investigated the effects of KillerRed-based PDT on tumor growth in vivo and examined the subsequent tumor metabolic states including the changes of nicotinamide adenine dinucleotide hydrogen(NADH)and flavoprotein(Fp),two important metabolic coenzymes of tumor *** showed the tumor growth had been significantly inhibited by KillerRedbased PDT treatment compared to control groups.A home-made cryo-imaging redox scanner was used to measure intrinsic fluorescence and exogenous KillerRed fluorescence signals in *** Fp signal was elevated by nearly 4.5-fold,while the NADH signal decreased by 66%after light irradiation,indicating that Fp and NADH were oxidized in the course of KillerRedbased ***,we also observed correlation between the fluorescence distribution of KillerRed and *** suggests that the KillerRed protein based PDT might provide a new approach for tumor therapy accompanied by altering tumor metabolism.

关键词： Reactive oxygen species redox metabolism flavoprotein nicotinamide adenine dinucleotide hydrogen

3D imaging of the mitochondrial redox state of rathearts under normal and fasting conditions

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Journal of Innovative Optical Health Sciences 2014年第2期7卷 60-69页

作者： He N.Xu Rong Zhou Lily Moon Min Feng Lin Z.Li department of radiology Universityof PennsyluaniaPhilade lphia PA 19104USA Briton chance laboratory of Redor imaging Johnson Research Foundation Department of Biochemistry and Biophysics University of Pennsyluania Philade lphiaPA 19104USA Institute of Translational Medicine and Therapeutics Perelman School of Medicine University of Pennsylvania Philadelphia PA 19104USA

The heart requires continuous ATP availability that is generated in the *** using the cell culture and perfiused organ models have been carried out to investigate thebiochemistry in the mitochondria in response to a change in substrate supply,mitochondrialbioenergetics of heart under normal feed or fasting conditions has not been studied at the tissuelevel with a sub-millimeter spatial resolution either in vivo or er *** of many food-derived metabolites to generate ATP in the mitochondria is realized through the NADH/NAD+couple acting as a central electron *** employed the chance redox scanner thelow-temperat ure fluorescence scanner to image the three-dimensional(3D)spatial distribution of themitochondrial redox states in heart tissues of rats under normai feeding or an overnight star-vation for 14.5 *** consecutive sections of each heart were imaged to map three redoxindices,i.e,NADH,oxidized favoproteins Fp,including flavin adenine dinucleotide(FAD)andthe redox ratio NADH/*** imaging results revealed the micro-heterogeneity and the spatial distribution of these redox *** quantitative analysis showed that in the fasted hearts thestandard deviation of both NADH and Fp,ie.,SD NADH and SDFp,significantly decreasedwith a p value of 0.032 and 0.045,respectively,indicating that the hearts become relatively morehomogeneous after *** fasted hearts contained 28.6%less NADH(p=0.038).No *** change in Fp was foumnd(p=0.4).The NADH/Fp ratio decreased with a marginalP value(0.076).The decreased NADH im the fasted hearts is consistent with the cardiac celsreliance of fatty acids consumption for energy metabolism when glucose becomes *** o bservation of N ADH decrease induced by dietary restriction in the heart at tissuelevel has not been reported to our best *** chance redox scanner demonstrated thefeasibility of 3D imaging of the mitochondrial redox st ate in the heart and

关键词： Mitochondion metabolism bioenergetics cardiomyocyte NADH flavoproteins Fp FAD caloric restriction food deprivation.

QUANTITATIVE redox SCANNING OF TISSUE SAMPLES USING A CALIBRATION PROCEDURE

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Journal of Innovative Optical Health Sciences 2009年第4期2卷 375-385页

作者： HE N.XU BAOHUA WU SHOKO NIOKA britton chance LIN Z.LI Molecular imaging laboratory Department of Radiology University of PennsylvaniaPhiladelphia PA 19104USA Johnson Research Foundation department of Biochemistry and Biophysics University of Pennsylvania PA 19104USA

The fluorescence properties of reduced nicotinamide adenine dinucleotide(NADH)and oxidizedflavoproteins(Fp)including flavin adenine dinucleotide(FAD)in the respiratory chain are sensitive indicators of intracellular metabolic states and have been applied to the studies of mitochondrial function with energy-linked *** redox scanner,a three-dimensional(3D)low temperature imager previously developed by chance et al.,measures the in vivo metabolicproperties of tissue samples by acquiring fluorescence images of NADH and *** redox ratios,***/(Fp+NADH)and NADH/(Fp+NADH),provided a sensitive index of the mitochondrialredox state and were determined based on relative signal intensity *** we report thefurther development of the redox scanning technique by using a calibration method to quantifythe nominal concentration of the fluorophores in *** redox scanner exhibited very goodlinear response in the range of NADH concentration between 165–1318µM and Fp between90–720µM using snap-frozen solution *** samples such as human tumor mousexenografts and various mouse organs were redox-scanned together with adjacent NADH and Fpstandards of known concentration at liquid nitrogen *** nominal NADH and Fpconcentrations as well as the redox ratios in the tissue samples were quantified by normalizing the tissue NADH and Fp fluorescence signal to that of the snap-frozen solution *** calibration procedure allows comparing redox images obtained at different time,independent of instrument *** quantitative multi-slice redox images revealed heterogeneity inmitochondrial redox state in the tissues.

关键词： NADH flavoprotein redox ratio mitochondria tissue heterogeneity