Spatial–temporal FUCCI imaging of each cell in a tumor demonstrates locational dependence of cell cycle dynamics and chemoresponsiveness

Yano, Shuya; Zhang, Yong; Miwa, Shinji; Tome, Yasunori; Hiroshima, Yukihiko; Uehara, Fuminari; Yamamoto, Masaaki; Suetsugu, Atsushi; Kishimoto, Hiroyuki; Tazawa, Hiroshi; Zhao, Ming; Bouvet, Michael; Fujiwara, Toshiyoshi; Hoffman, Robert M.

doi:10.4161/cc.29156

Cited by 68 publications

(83 citation statements)

References 37 publications

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“…2 The results also suggest that drugs that target quiescent cancer cells are urgently needed. 1 In the present study, we demonstrate that dormant/quiescent cancer cells induce nascent tumor vessels after chemotherapy, allowing tumors to regrow rapidly after cessation of treatment. Chemotherapy-treated tumors had much more and deeper tumor vessels than control tumors and rapidly began regrowing after cessation of treatment.…”

Section: Introductionmentioning

confidence: 80%

“…1). 1 A tumor 7 d after implantation consisted of 15.3 § 2.4% quiescent cancer cells and 84.7 § 2.4% proliferating cancer cells. A tumor 14 d after implantation consisted of 52.4 § 3.5% quiescent cancer cells and 47.6 § 3.5% proliferating cancer cells.…”

Section: Resultsmentioning

confidence: 99%

“…12 Plasmids expressing mKO2-hCdt1 (red/orange fluorescent protein) or mAG-hGem (green fluorescent protein) were obtained from the Medical and Biological Laboratory (City, Japan). 1 …”

Section: Establishment Of Mkn45 Cells Stably Transfected With Fucci-vmentioning

confidence: 99%

“…1 Nestin-driven green fluorescent protein (ND-GFP) transgenic nude mice carry the GFP gene under the control of the nestin promoter, were also bred and maintained at AntiCancer Inc. 3,[13][14][15] All animal studies were conducted in accordance with the principles and procedures outlined in the National Institute of Health Guide for the Care and Use of Animals under Assurance Number A3873-1.…”

Section: Animal Experimentsmentioning

confidence: 99%

“…These results indicate why most drugs currently in clinical use, which target cancer cells in S/G 2 /M, are mostly ineffective on solid tumors. 1 We have termed this phenomena tumor intrinsic chemoresistance (TIC). 2 The results also suggest that drugs that target quiescent cancer cells are urgently needed.…”

Section: Introductionmentioning

confidence: 99%

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Cell-cycle-dependent drug-resistant quiescent cancer cells induce tumor angiogenesis after chemotherapy as visualized by real-time FUCCI imaging

et al. 2017

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We previously demonstrated that quiescent cancer cells in a tumor are resistant to conventional chemotherapy as visualized with a fluorescence ubiquitination cell cycle indicator (FUCCI). We also showed that proliferating cancer cells exist in a tumor only near nascent vessels or on the tumor surface as visualized with FUCCI and green fluorescent protein (GFP)-expressing tumor vessels. In the present study, we show the relationship between cellcycle phase and chemotherapy-induced tumor angiogenesis using in vivo FUCCI real-time imaging of the cell cycle and nestin-driven GFP to detect nascent blood vessels. We observed that chemotherapy-treated tumors, consisting of mostly of quiescent cancer cells after treatment, had much more and deeper tumor vessels than untreated tumors. These newly-vascularized cancer cells regrew rapidly after chemotherapy. In contrast, formerly quiescent cancer cells decoyed to S/G 2 phase by a telomerase-dependent adenovirus did not induce tumor angiogenesis. The present results further demonstrate the importance of the cancer-cell position in the cell cycle in order that chemotherapy be effective and not have the opposite effect of stimulating tumor angiogenesis and progression.

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Section: Introductionmentioning

confidence: 80%

Section: Resultsmentioning

confidence: 99%

“…12 Plasmids expressing mKO2-hCdt1 (red/orange fluorescent protein) or mAG-hGem (green fluorescent protein) were obtained from the Medical and Biological Laboratory (City, Japan). 1 …”

Section: Establishment Of Mkn45 Cells Stably Transfected With Fucci-vmentioning

confidence: 99%

Section: Animal Experimentsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Cell-cycle-dependent drug-resistant quiescent cancer cells induce tumor angiogenesis after chemotherapy as visualized by real-time FUCCI imaging

et al. 2017

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Engineered bacteria detect spatial profiles in glucose concentration within solid tumor cell masses

Panteli

Forbes

2016

Biotech & Bioengineering

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Tumor heterogeneity makes cancer difficult to treat. Many small molecule cancer drugs target rapidly dividing cells on the periphery of tumors but have difficulty in penetrating deep into tumors and are ineffective at treating entire tumors. Targeting both rapidly dividing and slower growing regions of tumors is essential to effectively treat cancer. A cancer drug carrier that penetrates deep into tumors and identifies metabolically activity could supply treatment to those areas based on the local microenvironment. We hypothesized that glucose sensing bacteria could identify sugar gradients in solid tumors. To test this hypothesis, a genetic circuit was designed to trigger expression of a green fluorescent protein (GFP) reporter through the chemotaxis-osmoporin fusion protein, Trz1, a receptor for sensing glucose and ribose sugars. E. coli equipped with the Trz1-GFP expression system, were administered to an in vitro model of a continuously perfused tumor tissue that mimics systemic delivery and clearance of bacteria through a blood vessel adjacent to a solid tumor. The level of GFP expressed, per bacterium, was time independent and indicated the glucose concentration as a function of penetration depth within the microfluidic tumors. The measured glucose concentration, correlated (P-value = 2.6×10−5) with tumor cell viability as a function of depth. Mathematical analysis predicted drug delivery by glucose-sensing bacteria would eliminate a higher percentage of the viable tumor cell population than a systemically administered drug. Glucose-sensing bacteria could deliver cancer therapies with increased drug penetration and nutrient-dependent dosing to continuously treat viable regions of cancer tissue that have a higher prevalence for metastatic dissemination.

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CHO cells engineered for fluorescence read out of cell cycle and growth rate in real time

Fuge

Hong

Riecken

et al. 2017

Biotechnology Progress

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For efficient production of recombinant proteins by mammalian cells in a bioreactor, optimal growth rates are required and represent the most important process parameter. We present the first successful attempt to monitor the growth behavior and cell cycle state of a mammalian production relevant cell line under bioreactor cultivation conditions up to 1.2 l, utilizing a fluorescent read-out without the need of additional staining or marking. For this purpose, we developed two new production relevant cell line derivatives (CHO-K1 FUCCI CM & CHO-K1 FUCCI CN) and corresponding analytical methods. The approach is easily scalable, applicable to mammalian recombinant protein production cell lines, and it allows for real-time monitoring using appropriate fluorescence probes. It is based on the Ubiquitination-based Cell Cycle Indicator (FUCCI) system developed by Miyawaki et al. CHO-K1 was chosen as a model cell line due to its close relationship to several production cell lines. We defined a new process parameter i , a quantitative and numerically robust representation of the cell cycle distribution, and demonstrate it to be linearly correlated with the cell cycle state and inversely related to the real time growth rate. Detection of growth rate limitations is possible earlier than using cell-count-based approaches. Analytics were compatible with bulk fluorescence methods, using a plate reader as well as a flow cytometer. For future real time applications in industry scale bioreactors we recommend the use of on-line or at-line fluorescence probes. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1408-1417, 2017.

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Spatial–temporal FUCCI imaging of each cell in a tumor demonstrates locational dependence of cell cycle dynamics and chemoresponsiveness

Cited by 68 publications

References 37 publications

Cell-cycle-dependent drug-resistant quiescent cancer cells induce tumor angiogenesis after chemotherapy as visualized by real-time FUCCI imaging

Cell-cycle-dependent drug-resistant quiescent cancer cells induce tumor angiogenesis after chemotherapy as visualized by real-time FUCCI imaging

Engineered bacteria detect spatial profiles in glucose concentration within solid tumor cell masses

CHO cells engineered for fluorescence read out of cell cycle and growth rate in real time

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