Illuminating NAD+ Metabolism in Live Cells and In Vivo Using a Genetically Encoded Fluorescent Sensor

Zou, Yejun; Wang, Aoxue; Huang, Li; Zhu, Xudong; Hu, Qingmin; Zhang, Yinan; Chen, Xianjun; Li, Fengwen; Wang, Qiaohui; Wang, Hu; Liu, Renmei; Zuo, Fangting; Li, Ting; Yao, Jing; Qian, Yanlong; Shi, M.; Xiao, Yue; Chen, Weicai; Zhang, Zhuo; Wang, Congrong; Zhou, Yong; Zhu, Linyong; Ju, Zhenyu; Loscalzo, Joseph; Yang, Yi; Zhao, Yuzheng

doi:10.1016/j.devcel.2020.02.017

Cited by 75 publications

(81 citation statements)

References 54 publications

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“…In this work, we uncover how inter-cellular heterogeneity in metabolic state in breast cancer results from the combined variation in PI3K signaling, chromatin state, and actin dynamics ( Figures S7 H and S7I). Fluorescent biosensors have previously been used to perform high-content screens for metabolic perturbations, as well as interrogate metabolic features in vivo ; however, one under-explored feature that these tools provide is the ability to perform longitudinal single-cell analysis ( Takanaga et al., 2008 ; Zhao et al., 2015 ; Zou et al., 2020 ). Previous work using multiplex imaging of an AKT biosensor and AMPK FRET biosensor has shown that the cellular energetic state is synchronized with AKT activity ( Hung et al., 2017 ).…”

Section: Discussionmentioning

confidence: 99%

Single-cell resolved imaging reveals intra-tumor heterogeneity in glycolysis, transitions between metabolic states, and their regulatory mechanisms

et al. 2021

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Summary Inter-cellular heterogeneity in metabolic state has been proposed to influence many cancer phenotypes, including responses to targeted therapy. Here, we track the transitions and heritability of metabolic states in single PIK3CA mutant breast cancer cells, identify non-genetic glycolytic heterogeneity, and build on observations derived from methods reliant on bulk analyses. Using fluorescent biosensors in vitro and in tumors, we have identified distinct subpopulations of cells whose glycolytic and mitochondrial metabolism are regulated by combinations of phosphatidylinositol 3-kinase (PI3K) signaling, bromodomain activity, and cell crowding effects. The actin severing protein cofilin, as well as PI3K, regulates rapid changes in glucose metabolism, whereas treatment with the bromodomain inhibitor slowly abrogates a subpopulation of cells whose glycolytic activity is PI3K independent. We show how bromodomain function and PI3K signaling, along with actin remodeling, independently modulate glycolysis and how targeting these pathways affects distinct subpopulations of cancer cells.

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Section: Discussionmentioning

confidence: 99%

Single-cell resolved imaging reveals intra-tumor heterogeneity in glycolysis, transitions between metabolic states, and their regulatory mechanisms

et al. 2021

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“…In addition to Ca 2+ sensors, cpFP-based sensors to detect cofactors [ 94 ], cAMP [ 95 ], ATP [ 96 , 97 ], or neurotransmitters such as glutamate and GABA [ 98 , 99 ] were developed by inserting specific sensing domains to cp-FP. Different from these cytosolic cpFP-based sensors detecting diffusible signaling molecules, the cpFP-based voltage sensor, named ASAP, was designed by fusion of cpFP module to voltage-sensing domains tethered at plasma-membrane [ 100 , 101 ].…”

Section: Sensing Strategies Of Fp-based Biosensorsmentioning

confidence: 99%

Genetically Encoded Biosensors Based on Fluorescent Proteins

Kim

Lee

et al. 2021

Sensors

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Genetically encoded biosensors based on fluorescent proteins (FPs) allow for the real-time monitoring of molecular dynamics in space and time, which are crucial for the proper functioning and regulation of complex cellular processes. Depending on the types of molecular events to be monitored, different sensing strategies need to be applied for the best design of FP-based biosensors. Here, we review genetically encoded biosensors based on FPs with various sensing strategies, for example, translocation, fluorescence resonance energy transfer (FRET), reconstitution of split FP, pH sensitivity, maturation speed, and so on. We introduce general principles of each sensing strategy and discuss critical factors to be considered if available, then provide representative examples of these FP-based biosensors. These will help in designing the best sensing strategy for the successful development of new genetically encoded biosensors based on FPs.

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“…Several genetically encoded sensors able to detect the NADH/NAD + ratio in living cells have been reported [ 204 , 205 ], but the absolute quantification of NAD + has only recently been obtained by Cambronne et al [ 206 ] who reported a radiometric sensor based on bacterial DNA-ligase and cpVenusFP (LigA-cpVenus), showing fluorescence reduction upon NAD + binding. Very recently, Zou et al [ 207 ] managed to obtain a sensor lighting in response to NAD + (FiNAD). “FiNad” is based on the insertion of cpYFP into the NAD + /NADH binding domain of the bacterial transcription repressor protein (T-Rex), optimized in order to recognize specifically NAD + .…”

Section: Fluorescence-based Biosensorsmentioning

confidence: 99%

Emergent Biosensing Technologies Based on Fluorescence Spectroscopy and Surface Plasmon Resonance

Camarca

Varriale

Capo

et al. 2021

Sensors

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The purpose of this work is to provide an exhaustive overview of the emerging biosensor technologies for the detection of analytes of interest for food, environment, security, and health. Over the years, biosensors have acquired increasing importance in a wide range of applications due to synergistic studies of various scientific disciplines, determining their great commercial potential and revealing how nanotechnology and biotechnology can be strictly connected. In the present scenario, biosensors have increased their detection limit and sensitivity unthinkable until a few years ago. The most widely used biosensors are optical-based devices such as surface plasmon resonance (SPR)-based biosensors and fluorescence-based biosensors. Here, we will review them by highlighting how the progress in their design and development could impact our daily life.

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Illuminating NAD+ Metabolism in Live Cells and In Vivo Using a Genetically Encoded Fluorescent Sensor

Cited by 75 publications

References 54 publications

Single-cell resolved imaging reveals intra-tumor heterogeneity in glycolysis, transitions between metabolic states, and their regulatory mechanisms

Single-cell resolved imaging reveals intra-tumor heterogeneity in glycolysis, transitions between metabolic states, and their regulatory mechanisms

Genetically Encoded Biosensors Based on Fluorescent Proteins

Emergent Biosensing Technologies Based on Fluorescence Spectroscopy and Surface Plasmon Resonance

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