Generating Primary Fibroblast Cultures from Mouse Ear and Tail Tissues

Khan, Muznah; Gasser, Stephan

doi:10.3791/53565

Cited by 70 publications

(67 citation statements)

References 18 publications

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“…However, this kind of cell preparation would take a long time to reach senescence and only produce a finite number of cells for testing. Alternatively, using an in vitro mouse replicative senescence model is more feasible, as cells they stop proliferating after 5-6 passages (Khan and Gasser, 2016). Senolytics or senostatics could next be applied in vivo using aged wild-type mice.…”

Section: Conclusion: a Long Road Aheadmentioning

confidence: 99%

An Appraisal on the Value of Using Nutraceutical Based Senolytics and Senostatics in Aging

Kaur

Macip

Stover

2020

Front. Cell Dev. Biol.

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The average human life expectancy has increased globally, and continues to rise, owing to the substantive progress made in healthcare, medicine, sanitation, housing and education. This ultimately enriches society with a greater proportion of elderly people. Sustaining a healthy aged population is key to diminish the societal and economic impact of age-related infirmities. This is especially challenging because tissue function, and thus wellbeing, naturally progressively decline as humans age. With age increasing the risk of developing diseases, one of the therapeutic options is to interfere with the molecular and cellular pathways involved in age-related tissue dysfunction, which is in part caused by the accumulation of senescent cells. One strategy to prevent this could be using drugs that selectively kill these cells (senolytics). In parallel, some compounds have been identified that prevent or slow down the progression of senescence or some of its features (senostatics). Senolytic and senostatic therapies have been shown to be efficient in vivo, but they also have unwanted dose-dependent side effects, including toxicity. Important advances might be made using bioactive compounds from plants and foods (nutraceuticals) if, as is proposed, they offer similar effectiveness with fewer side effects. The focus of this review is on the use of nutraceuticals in interfering with cellular senescence.

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Section: Conclusion: a Long Road Aheadmentioning

confidence: 99%

An Appraisal on the Value of Using Nutraceutical Based Senolytics and Senostatics in Aging

Kaur

Macip

Stover

2020

Front. Cell Dev. Biol.

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“…I25BD461E) from the UK Home Office. Fibroblasts were prepared from Npc1 -/- and Npc1 +/+ mice according to published methods (Khan and Gasser, 2016).…”

Section: Experimental Model and Subject Detailsmentioning

confidence: 99%

Redistribution of cholesterol from vesicle to plasmalemma controls fusion pore geometry

Rituper

Guček

Lisjak

et al. 2020

Preprint

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Highlights 36  Intravesicular cholesterol redistributes to the outer leaflet of the plasmalemma. 37 3  Cholesterol depletion widens the fusion pore, whereas cholesterol enrichment constricts the fusion 38 pore. 39  A model of cholesterol-dependent force preventing fusion pore widening is developed. 40  Disease-related increase in vesicle cholesterol constricts the fusion pore. 41 ABSTRACT 42 Eukaryotic vesicles fuse with the plasmalemma to form the fusion pore, previously considered to be 43 unstable with widening of the pore diameter. Recent studies established that the pore diameter is 44 stable, reflecting balanced forces of widening and closure. Proteins are considered key regulators of 45 the fusion pore, whereas the role of membrane lipids remains unclear. Super-resolution microscopy 46 revealed that lactotroph secretory vesicles discharge cholesterol after stimulation of exocytosis; 47 subsequently, vesicle cholesterol redistributes to the outer leaflet of the plasmalemma. Cholesterol 48 depletion in lactotrophs and astrocytes evokes release of vesicle hormone, indicating that cholesterol 49 constricts the fusion pore. A new model of cholesterol-dependent fusion pore diameter regulation is 50 proposed. High-resolution measurements of fusion pore conductance confirmed that the fusion pore 51 widens with cholesterol depletion and constricts with cholesterol enrichment. In fibroblasts lacking 52 the Npc1 protein, in which cholesterol accumulates in vesicles, the fusion pore is narrower than in 53 controls, showing that cholesterol regulates fusion pore geometry. 54 65 to the membrane area, and is affected by vesicle fusion and fission (Neher and Marty, 1982).66

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“…To a 20 mL scintillation vial containing a magnetic stir bar under an Ar(g) atmosphere was added cobimetinib (238 mg, 0.45 mmol, 2.50 equiv. ), (1''S, 3''S)-dispiro[adamantane-2,3'- [1,2,4]trioxolane-5',1''-cyclohexan]-3''-yl (4-nitrophenyl)carbonate (80 mg, 0.18 mmol, 1.0 equiv), N,N-dimethylformamide (1.60 mL), N,N-diisopropylethylamine (78 uL, 0.45 mmol, 2.50 equiv), and 4-dimethylaminopyridine (28.6 mg, 0.23 mmol, 1.3 equiv). The reaction was stirred at rt for 12 h and monitored by TLC and LC/MS.…”

Section: Yl)piperidine-1-carboxylatementioning

confidence: 99%

“…The crude material was purified by preparative HPLC (40-100% MeOH-water + 0.05% formic acid) and the product-bearing fractions were combined, concentrated and lyophilized to give (1''S,3''S)-TRX-COBI (65.2 mg, 0.078 mmol, 43%) as a colorless lyophilized solid (9:1 dr, trans:cis). 1…”

Section: Yl)piperidine-1-carboxylatementioning

confidence: 99%

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Exploiting KRAS-driven Ferroaddiction in Cancer Through Ferrous Iron-Activatable Drug Conjugates (FeADC)

Jiang

Muir

Gonciarz

et al. 2020

Preprint

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KRAS mutations cause a quarter of cancer mortality and currently are not sensitive to any targeted, FDA-approved agents. Several inhibitors of the MAPK pathway are FDA approved but exhibit low clinical tolerability at doses needed to adequately extinguish KRAS signaling. We discovered an avidity for ferrous iron (Fe 2+ ) induced by and dependent on oncogenic KRAS signaling. We leveraged an FDA-approved MEK inhibitor to produce a prototypical Ferrous Iron-Activatable Drug Conjugate (FeADC) and with this novel agent achieved MAPK blockade in tumor cells while sparing normal tissues. These improvements allowed sustainable, effective treatment of tumor bearing animals with superior tolerability. Iron-activated therapeutics are unknown outside of antiparasitic therapy but may hold significant promise for the treatment of KRAS-driven solid tumors.driven PDA xenografts as compared to breast cancer xenografts with wt KRAS (Figure 1E). To assess levels of reduced iron in the same tumors, we used 18 F-TRX, our recently described (Muir et al., 2019) oxidation-state specific radiotracer for Fe 2+ . We found that the mKRAS-driven PDA tumors were enriched in Fe 2+ as compared to the wt KRAS tumors, mirroring our findings with 68 Ga (Figure S1A). Together these findings suggest that increased iron uptake and iron reduction are common in PDA and associated with poor patient prognosis.

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Generating Primary Fibroblast Cultures from Mouse Ear and Tail Tissues

Cited by 70 publications

References 18 publications

An Appraisal on the Value of Using Nutraceutical Based Senolytics and Senostatics in Aging

An Appraisal on the Value of Using Nutraceutical Based Senolytics and Senostatics in Aging

Redistribution of cholesterol from vesicle to plasmalemma controls fusion pore geometry

Exploiting KRAS-driven Ferroaddiction in Cancer Through Ferrous Iron-Activatable Drug Conjugates (FeADC)

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