Optical Metabolic Imaging Identifies Glycolytic Levels, Subtypes, and Early-Treatment Response in Breast Cancer

Walsh, Alex J.; Cook, Rebecca S.; Manning, H. Charles; Hicks, Donna J.; Lafontant, Alec; Arteaga, Carlos L.; Skala, Melissa C.

doi:10.1158/0008-5472.can-13-0527

Cited by 268 publications

(343 citation statements)

References 53 publications

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“…Phasor fluorescence lifetime microscopy (FLIM) analysis proceeds by Fourier transformation of the lifetime data, which allows direct quantification at each pixel of the free and bound NADH ratio (34)(35)(36). Metabolic transitions and phenotypes in living single cells have been previously determined using the phasor approach to FLIM, including identification of biological processes, such as cell proliferation and differentiation, tumorigenesis, and aging (22,24,27,37,38). Because biochemical reactions depending on NADH occur at different rates in distinct subcellular compartments, the measurement of NADH free/bound ratio provides a weighted mean of the enzymatic activities in the cell when combined with local NADH fluorescence intensity.…”

Section: Resultsmentioning

confidence: 99%

“…Lifetime measurements from fluorescent NADH distinguish free NADH and subpopulations of protein-bound NADH, whereas NAD + is not fluorescent (21). NADH 2P-FLIM cellular map provides sensitive measurements of local activity associated with NADH metabolism (22)(23)(24)(25)(26)(27)(28). Fluorescence correlation spectroscopy analyzes the fluctuation of fluorescent molecules in a small illuminated spot, providing spatiotemporal maps of concentration, interaction, or diffusion parameters of molecules (29)(30)(31)(32)(33).…”

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confidence: 99%

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Spatial dynamics of SIRT1 and the subnuclear distribution of NADH species

Aguilar-Arnal

Ranjit

Stringari

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Sirtuin 1 (SIRT1) is an NAD + -dependent deacetylase that functions as metabolic sensor of cellular energy and modulates biochemical pathways in the adaptation to changes in the environment. SIRT1 substrates include histones and proteins related to enhancement of mitochondrial function as well as antioxidant protection. Fluctuations in intracellular NAD + levels regulate SIRT1 activity, but how SIRT1 enzymatic activity impacts on NAD + levels and its intracellular distribution remains unclear. Here, we show that SIRT1 determines the nuclear organization of protein-bound NADH. Using multiphoton microscopy in live cells, we show that free and bound NADH are compartmentalized inside of the nucleus, and its subnuclear distribution depends on SIRT1. Importantly, SIRT6, a chromatin-bound deacetylase of the same class, does not influence NADH nuclear localization. In addition, using fluorescence fluctuation spectroscopy in single living cells, we reveal that NAD + metabolism in the nucleus is linked to subnuclear dynamics of active SIRT1. These results reveal a connection between NAD + metabolism, NADH distribution, and SIRT1 activity in the nucleus of live cells and pave the way to decipher links between nuclear organization and metabolism.S irtuins (SIRTs) are a conserved family of deacetylases that target a variety of proteins located in virtually all cellular compartments (1). Deacetylation by SIRTs may control many functional aspects of target proteins (2). Because SIRT deacetylase activity depends on the energy carrier NAD + , these enzymes are thought to operate as cellular metabolic sensors. In addition, because histones are targeted by nuclear SIRT, these enzymes could link variations in cellular metabolism to chromatin function.There are seven mammalian SIRTs (SIRT1 to SIRT7) with distinct subcellular locations. SIRT2 is mainly cytoplasmic; SIRT3, SIRT4, and SIRT5 are found in the mitochondrial compartment; and SIRT1, SIRT6, and SIRT7 are located in the cell nucleus (1). In mammals, SIRT1 contributes to development and protects from metabolic and cardiovascular disease, neurodegeneration, and cancer (3). SIRT1 has been reported to promote healthy aging and regulate lifespan (4, 5). At the cellular level, SIRT1 regulates lipid and glucose homeostasis, apoptosis, DNA repair, and mitochondrial function. Variations in NAD + levels control SIRT1 activity (6-9), a relevant finding in the regulation of circadian rhythms (10). Circadian rhythms in NAD + levels have been observed (11,12), which lead to fluctuating SIRT1 deacetylase activity (9) that, in turn, results into cyclic acetylation of specific SIRT1 targets (6, 9, 13). SIRT1 and SIRT6 segregate circadian metabolism by driving transcription of a differential subset of circadian genes (14). SIRT6 is a chromatinbound protein that was first characterized as a regulator of genome stability (15). The other nuclear SIRT, SIRT7, appears to be highly localized in the nucleolus and possibly involved in Pol-I-dependent transcription (16), and it has been shown to regula...

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Spatial dynamics of SIRT1 and the subnuclear distribution of NADH species

Aguilar-Arnal

Ranjit

Stringari

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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“…The histogram of photons by lifetime can be deconvolved from the system response and fit to an exponential decay function to resolve both short and long lifetimes, as well as the proportion of short and long lifetimes ( Fig. 2A) (35). (Additional details on fluorescence lifetime imaging can be found in a book by Marcu et al (36).…”

Section: Optical Imaging Of Organoidsmentioning

confidence: 99%

“…OMI uses multiphoton microscopy to probe the fluorescence intensities and lifetimes of NADH and FAD, coenzymes of cellular metabolism processes. OMI includes the optical redox ratio (the fluorescence intensity of NADH divided by the intensity of FAD), which provides a relative measure of the glycolytic state of the cell (35). Figures 2B-2D show representative OMI images of an organoid derived from a human breast cancer biopsy sample.…”

Section: Omi As An Early Measure Of Drug Response In Organoidsmentioning

confidence: 99%

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Functional Optical Imaging of Primary Human Tumor Organoids: Development of a Personalized Drug Screen

2017

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Primary tumor organoids are a robust model of individual human cancers and present a unique platform for patient-specific drug testing. Optical imaging is uniquely suited to assess organoid function and behavior because of its subcellular resolution, penetration depth through the entire organoid, and functional endpoints. Specifically, optical metabolic imaging (OMI) is highly sensitive to drug response in organoids, and OMI in tumor organoids correlates with primary tumor drug response. Therefore, an OMI organoid drug screen could enable accurate testing of drug response for individualized cancer treatment. The objective of this perspective is to introduce OMI and tumor organoids to a general audience in order to foster the adoption of these techniques in diverse clinical and laboratory settings.

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Patient‐derived organoids as a preclinical platform for precision medicine in colorectal cancer

et al. 2022

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Patient‐derived organoids are being considered as models that can help guide personalized therapy through in vitro anticancer drug response evaluation. However, attempts to quantify in vitro drug responses in organoids and compare them with responses in matched patients remain inadequate. In this study, we investigated whether drug responses of organoids correlate with clinical responses of matched patients and disease progression of patients. Organoids were established from 54 patients with colorectal cancer who (except for one patient) did not receive any form of therapy before, and tumor organoids were assessed through whole‐exome sequencing. For comparisons of in vitro drug responses in matched patients, we developed an ‘organoid score’ based on the variable anticancer treatment responses observed in organoids. Very interestingly, a higher organoid score was significantly correlated with a lower tumor regression rate after the standard‐of‐care treatment in matched patients. Additionally, we confirmed that patients with a higher organoid score (≥ 2.5) had poorer progression‐free survival compared with those with a lower organoid score (< 2.5). Furthermore, to assess potential drug repurposing using an FDA‐approved drug library, ten tumor organoids derived from patients with disease progression were applied to a simulation platform. Taken together, organoids and organoid scores can facilitate the prediction of anticancer therapy efficacy, and they can be used as a simulation model to determine the next therapeutic options through drug screening. Organoids will be an attractive platform to enable the implementation of personalized therapy for colorectal cancer patients.

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Optical Metabolic Imaging Identifies Glycolytic Levels, Subtypes, and Early-Treatment Response in Breast Cancer

Cited by 268 publications

References 53 publications

Spatial dynamics of SIRT1 and the subnuclear distribution of NADH species

Spatial dynamics of SIRT1 and the subnuclear distribution of NADH species

Functional Optical Imaging of Primary Human Tumor Organoids: Development of a Personalized Drug Screen

Patient‐derived organoids as a preclinical platform for precision medicine in colorectal cancer

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