A quantitative FastFUCCI assay defines cell cycle dynamics at a single-cell level

Koh, Siang-Boon; Mascalchi, Patrice; Rodríguez, Esther; Lin, Yao; Jodrell, Duncan I.; Richards, Frances M.; Lyons, Scott K.

doi:10.1242/jcs.195164

Cited by 76 publications

(84 citation statements)

References 39 publications

(45 reference statements)

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“…5A). This may explain why it was previously found that the interference with the DNA replication origin activity enhanced the response of cells to paclitaxel (16).…”

Section: Discussionmentioning

confidence: 91%

“…While the blockage in S phase is expected after gemcitabine exposure, due to depletion of the nucleotide pool required for DNA duplication, cells treated with nab-paclitaxel were expected to arrest in mitosis or late G2 phase due to defects in spindle elongation. However, recent data have revealed that cells treated with paclitaxel often proceeded through mitosis into the next interphase, where the majority of cell deaths occurred (16). In particular, nab-paclitaxel seemed to interfere with the very early stages of the S phase in PDAC cells (Fig.…”

Section: Discussionmentioning

confidence: 93%

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Co-treatment with gemcitabine and nab-paclitaxel exerts additive effects on pancreatic cancer cell death

Passacantilli

Panzeri²,

Terracciano³

et al. 2018

Oncol Rep

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Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive cancer and current treatments exert small effects on life expectancy. The most common adjuvant treatment for PDAC is gemcitabine. However, relapse almost invariably occurs and most patients develop metastatic, incurable disease. The aim of the present study was to assess the activity of nanoparticle albumin-bound paclitaxel (nab-paclitaxel) alone or in combination with gemcitabine in PDAC cell lines displaying different degrees of sensitivity to gemcitabine treatment. We evaluated the effects of gemcitabine and nab-paclitaxel and their combination on cell proliferation, death, apoptosis and cell cycle distribution in PDAC cell lines either sensitive to gemcitabine, or with primary or secondary resistance to gemcitabine. Our results indicated that the dose‑response of PDAC cell lines to nab-paclitaxel was similar, regardless of their sensitivity to gemcitabine. In addition, nab-paclitaxel elicited similar cytotoxic effects on a PDAC cell line highly resistant to gemcitabine that was selected after prolonged exposure to the drug. Notably, we found that combined treatment with gemcitabine and nab-paclitaxel exerted additive effects on cell death, even at lower doses of the drugs. The combined treatment caused an increase in cell death by apoptosis and in cell cycle blockage in S phase, as assessed by flow cytometry and western blot analysis of the PARP-1 cleavage. These results revealed that a combined treatment with nab-paclitaxel may overcome resistance to gemcitabine and may represent a valuable therapeutic approach for PDAC.

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“…5A). This may explain why it was previously found that the interference with the DNA replication origin activity enhanced the response of cells to paclitaxel (16).…”

Section: Discussionmentioning

confidence: 91%

Section: Discussionmentioning

confidence: 93%

Co-treatment with gemcitabine and nab-paclitaxel exerts additive effects on pancreatic cancer cell death

Passacantilli

Panzeri²,

Terracciano³

et al. 2018

Oncol Rep

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“…A problem with the original FUCCI was the need to use two separate plasmids, which may not be equally expressed, thereby confusing the color specificity of the various cell-cycle phases. FastFUCCI was subsequently developed, which has an all-in-one expression cassette that has all FUCCI genes under the control of a single promoter, resulting in their equal expression; these different FUCCI systems are compared in Figure 2 [21].…”

Section: Fucci Images Cell-cycle Dynamics Of Cancer Cells In Real-timementioning

confidence: 99%

FUCCI Real-Time Cell-Cycle Imaging as a Guide for Designing Improved Cancer Therapy: A Review of Innovative Strategies to Target Quiescent Chemo-Resistant Cancer Cells

Yano

Tazawa

Kagawa

et al. 2020

Cancers

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Progress in chemotherapy of solid cancer has been tragically slow due, in large part, to the chemoresistance of quiescent cancer cells in tumors. The fluorescence ubiquitination cell-cycle indicator (FUCCI) was developed in 2008 by Miyawaki et al., which color-codes the phases of the cell cycle in real-time. FUCCI utilizes genes linked to different color fluorescent reporters that are only expressed in specific phases of the cell cycle and can, thereby, image the phases of the cell cycle in real-time. Intravital real-time FUCCI imaging within tumors has demonstrated that an established tumor comprises a majority of quiescent cancer cells and a minor population of cycling cancer cells located at the tumor surface or in proximity to tumor blood vessels. In contrast to most cycling cancer cells, quiescent cancer cells are resistant to cytotoxic chemotherapy, most of which target cells in S/G2/M phases. The quiescent cancer cells can re-enter the cell cycle after surviving treatment, which suggests the reason why most cytotoxic chemotherapy is often ineffective for solid cancers. Thus, quiescent cancer cells are a major impediment to effective cancer therapy. FUCCI imaging can be used to effectively target quiescent cancer cells within tumors. For example, we review how FUCCI imaging can help to identify cell-cycle-specific therapeutics that comprise decoy of quiescent cancer cells from G1 phase to cycling phases, trapping the cancer cells in S/G2 phase where cancer cells are mostly sensitive to cytotoxic chemotherapy and eradicating the cancer cells with cytotoxic chemotherapy most active against S/G2 phase cells. FUCCI can readily image cell-cycle dynamics at the single cell level in real-time in vitro and in vivo. Therefore, visualizing cell cycle dynamics within tumors with FUCCI can provide a guide for many strategies to improve cell-cycle targeting therapy for solid cancers.

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“…Further investigations into the field of stem cell differentiation may yield more clinically applicable therapeutics. Mouse models with fluorescence ubiquitination cell cycle indicator (FUCCI) markers (Zielke and Edgar, 2015; Koh et al, 2017) and Lgr5-GFP positive cells (Bramhall et al, 2014), have been used to find small molecules capable of inducing stem cell differentiation into hair cell progenitors. Through the use of these models, a small molecule CHIR99021, a GSK inhibitor capable of inducing cochlear progenitor cells has been identified (Roccio et al, 2015).…”

Section: Specialized Sensory Cell Regenerationmentioning

confidence: 99%

Recent Advancements in the Regeneration of Auditory Hair Cells and Hearing Restoration

Mittal

Nguyen

Patel

et al. 2017

Front. Mol. Neurosci.

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Neurosensory responses of hearing and balance are mediated by receptors in specialized neuroepithelial sensory cells. Any disruption of the biochemical and molecular pathways that facilitate these responses can result in severe deficits, including hearing loss and vestibular dysfunction. Hearing is affected by both environmental and genetic factors, with impairment of auditory function being the most common neurosensory disorder affecting 1 in 500 newborns, as well as having an impact on the majority of elderly population. Damage to auditory sensory cells is not reversible, and if sufficient damage and cell death have taken place, the resultant deficit may lead to permanent deafness. Cochlear implants are considered to be one of the most successful and consistent treatments for deaf patients, but only offer limited recovery at the expense of loss of residual hearing. Recently there has been an increased interest in the auditory research community to explore the regeneration of mammalian auditory hair cells and restoration of their function. In this review article, we examine a variety of recent therapies, including genetic, stem cell and molecular therapies as well as discussing progress being made in genome editing strategies as applied to the restoration of hearing function.

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A quantitative FastFUCCI assay defines cell cycle dynamics at a single-cell level

Cited by 76 publications

References 39 publications

Co-treatment with gemcitabine and nab-paclitaxel exerts additive effects on pancreatic cancer cell death

Co-treatment with gemcitabine and nab-paclitaxel exerts additive effects on pancreatic cancer cell death

FUCCI Real-Time Cell-Cycle Imaging as a Guide for Designing Improved Cancer Therapy: A Review of Innovative Strategies to Target Quiescent Chemo-Resistant Cancer Cells

Recent Advancements in the Regeneration of Auditory Hair Cells and Hearing Restoration

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