Cellular Senescence in Vitro

In 1960, Rita Levi-Montalcini and Barbara Booker made an observation that transformed neuroscience: as neurons mature, they become apoptosis resistant. The following year Leonard Hayflick and Paul Moorhead described a stable replicative arrest of cells in vitro, termed “senescence”. For nearly 60 years, the cell biology fields of neuroscience and senescence ran in parallel, each separately defining phenotypes and uncovering molecular mediators to explain the 1960s observations of their founding mothers and fathers, respectively. During this time neuroscientists have consistently observed the remarkable ability of neurons to survive. Despite residing in environments of chronic inflammation and degeneration, as occurs in numerous neurodegenerative diseases, often times the neurons with highest levels of pathology resist death. Similarly, cellular senescence (hereon referred to simply as “senescence”) now is recognized as a complex stress response that culminates with a change in cell fate. Instead of reacting to cellular/DNA damage by proliferation or apoptosis, senescent cells survive in a stable cell cycle arrest. Senescent cells simultaneously contribute to chronic tissue degeneration by secreting deleterious molecules that negatively impact surrounding cells. These fields have finally collided. Neuroscientists have begun applying concepts of senescence to the brain, including post-mitotic cells. This initially presented conceptual challenges to senescence cell biologists. Nonetheless, efforts to understand senescence in the context of brain aging and neurodegenerative disease and injury emerged and are advancing the field. The present review uses pre-defined criteria to evaluate evidence for post-mitotic brain cell senescence. A closer interaction between neuro and senescent cell biologists has potential to advance both disciplines and explain fundamental questions that have plagued their fields for decades.

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Calcium channel ITPR2 and mitochondria–ER contacts promote cellular senescence and aging

Nature Communications ◽

10.1038/s41467-021-20993-z ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Dorian V. Ziegler ◽

David Vindrieux ◽

Delphine Goehrig ◽

Sara Jaber ◽

Guillaume Collin ◽

...

Keyword(s):

Cellular Senescence ◽

Calcium Release ◽

Liver Steatosis ◽

Metabolic Stress ◽

Calcium Release Channel ◽

Release Channel ◽

Age Related ◽

Trisphosphate Receptor

AbstractCellular senescence is induced by stresses and results in a stable proliferation arrest accompanied by a pro-inflammatory secretome. Senescent cells accumulate during aging, promoting various age-related pathologies and limiting lifespan. The endoplasmic reticulum (ER) inositol 1,4,5-trisphosphate receptor, type 2 (ITPR2) calcium-release channel and calcium fluxes from the ER to the mitochondria are drivers of senescence in human cells. Here we show that Itpr2 knockout (KO) mice display improved aging such as increased lifespan, a better response to metabolic stress, less immunosenescence, as well as less liver steatosis and fibrosis. Cellular senescence, which is known to promote these alterations, is decreased in Itpr2 KO mice and Itpr2 KO embryo-derived cells. Interestingly, ablation of ITPR2 in vivo and in vitro decreases the number of contacts between the mitochondria and the ER and their forced contacts induce premature senescence. These findings shed light on the role of contacts and facilitated exchanges between the ER and the mitochondria through ITPR2 in regulating senescence and aging.

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Pigment epithelium-derived factor delays cellular senescence of human mesenchymal stem cells in vitro by reducing oxidative stress

Cell Biology International ◽

10.1002/cbin.10041 ◽

2013 ◽

Vol 37 (4) ◽

pp. 305-313 ◽

Cited By ~ 6

Author(s):

Yukun Cao ◽

Ting Yang ◽

Chunhu Gu ◽

Dinghua Yi

Keyword(s):

Oxidative Stress ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Cellular Senescence ◽

Pigment Epithelium ◽

Human Mesenchymal Stem Cells ◽

Pigment Epithelium Derived Factor

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Senolytic Therapy for Cerebral Ischemia-Reperfusion Injury

International Journal of Molecular Sciences ◽

10.3390/ijms222111967 ◽

2021 ◽

Vol 22 (21) ◽

pp. 11967

Author(s):

Songhyun Lim ◽

Tae Jung Kim ◽

Young-Ju Kim ◽

Cheesue Kim ◽

Sang-Bae Ko ◽

...

Keyword(s):

Ischemic Stroke ◽

Cerebral Ischemia ◽

Inflammatory Cytokines ◽

Cellular Senescence ◽

Therapeutic Potential ◽

Ischemia Reperfusion ◽

Severe Disabilities ◽

Cerebral Ischemia Reperfusion ◽

Cerebral Ir Injury

Ischemic stroke is one of the leading causes of death, and even timely treatment can result in severe disabilities. Reperfusion of the ischemic stroke region and restoration of the blood supply often lead to a series of cellular and biochemical consequences, including generation of reactive oxygen species (ROS), expression of inflammatory cytokines, inflammation, and cerebral cell damage, which is collectively called cerebral ischemia-reperfusion (IR) injury. Since ROS and inflammatory cytokines are involved in cerebral IR injury, injury could involve cellular senescence. Thus, we investigated whether senolytic therapy that eliminates senescent cells could be an effective treatment for cerebral IR injury. To determine whether IR induces neural cell senescence in vitro, astrocytes were subjected to oxygen-glucose deprivation/reoxygenation (OGD/R). OGD/R induced astrocyte senescence and senescent cells in OGD/R-injured astrocytes were effectively eliminated in vitro by ABT263, a senolytic agent. IR in rats with intraluminal middle cerebral artery occlusion induced cellular senescence in the ischemic region. The senescent cells in IR-injured rats were effectively eliminated by intravenous injections of ABT263. Importantly, ABT263 treatment significantly reduced the infarct volume and improved neurological function in behavioral tests. This study demonstrated, for the first time, that senolytic therapy has therapeutic potential for cerebral IR injury.

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Μελέτη της κυτταρικής γήρανσης στον ομαλό λειχήνα του στόματος με τη χρήση της ειδικής χρώσης της λιποφουσκίνης με sudan black B, μιά πρωτότυπη μέθοδο ανίχνευσης γηρασμένων κυττάρων

10.12681/eadd/41503 ◽

2014 ◽

Author(s):

Ελένη Γεωργακοπούλου

Keyword(s):

Cellular Senescence ◽

Replicative Senescence ◽

Sudan Black B ◽

Beta Galactosidase

Ως κυτταρική γήρανση (cellular senescence) ορίζεται η μη αναστρέψιμη παύση του κυτταρικού κύκλου συνεπεία είτε εξάντλησης των τελομερών, είτε κυτταρικού στρες, και θεωρείται μηχανισμός αντίστασης στην καρκινογένεση. Το φαινόμενο της κυτταρικής γήρανσης αποτελεί μια ερευνητική πρόκληση μιας και αποτελεί συνδετικό κρίκο μεταξύ της φυσιολογικής γήρανσης της χρόνιας φλεγμονής και βασικών μονοπατιών της καρκινογένεσης. Ο μέχρι σήμερα πιο αξιόπιστος δείκτης κυτταρικής γήρανσης είναι η ανίχνευση της δραστηριότητας της β- γαλακτοσιδάσης των γηρασμένων κυττάρων ,(senescence-associated-beta-galactosidase SA-β-gal). Η μέθοδος αυτή δεν μπορεί να εφαρμοστεί σε αρχειακό υλικό (ιστούς εγκιβωτισμένους σε παραφίνη) αλλά μόνο σε φρέσκους ιστούς και σε τομές από άμεσα κατεψυγμένους ιστούς (κρυοτομές). Εξαιτίας αυτού του περιορισμού υπάρχει έλλειψη εκτενών κλινικοπαθολογικών μελετών για την κυτταρική γήρανση.ΣΚΟΠΟΣ : Επιχειρήθηκε η αναζήτηση ενός βιολογικού δείκτη κυτταρικής γήρανσης, με εφαρμογή σε αρχειακό υλικό. Επίσης, μελετήθηκε η πρωτότυπη χρησιμοποίησή του ως δείκτη κυτταρικής γήρανσης σε ένα χρόνιο φλεγμονώδες νόσημα που αποτελεί μοντέλο συσχέτισης χρόνιας φλεγμονής και καρκίνου, τον Ομαλό λειχήνα του στόματος.ΥΛΙΚΑ ΚΑΙ ΜΕΘΟΔΟΙ : Αναζητώντας μια μέθοδο ανίχνευσης γηρασμένων κυττάρων εφαρμόσιμη σε αρχειακό υλικό, αξιολογήσαμε την ιστοχημική χρώση Sudan-Black B (SBB), που είναι ειδική για την ανίχνευση της λιποφουσκίνης. Η λιποφουσκίνη είναι ένα συσσωμάτωμα οξειδωμένων πρωτεϊνών, λιπιδίων και μετάλλων, η οποία συσσωρεύεται σε γηρασμένους ιστούς. Αναλύσαμε κυτταρικά συστήματα στα οποία προκλήθηκε κυτταρική γήρανση είτε ύστερα από εξάντληση του πολλαπλασιασμού (replicative senescence) ή από στρεσογόνα ερεθίσματα, προ-καρκινικές αλλοιώσεις σε υπό προϋποθέσεις knock-in ποντίκια που παρουσιάζουν γήρανση, και τέλος σε ανθρώπινες προκαρκινικές αλλοιώσεις που γνωρίζουμε ότι περιέχουν γηρασμένα κύτταρα. Η τεχνική εν συνεχεία εφαρμόστηκε σε δείγματα Ομαλού λειχήνα, Ακανθοκυτταρικού καρκινώματος στόματος , καλοήθεις βλάβες στοματικού βλεννογόνου (ινώματα) και φυσιολογικού στοματικού βλεννογόνου.ΑΠΟΤΕΛΕΣΜΑΤΑ : Στα παραπάνω πειράματα αποδείξαμε την συνταύτιση της λιποφουσκίνης και του SA-β-gal σε in vitro και in vivo γηρασμένα κύττταρα (κατεψυγμένους ιστούς). Tα ευρήματα αυτά συνηγορούν πως η λιποφουσκίνη είναι ένας ικανός υποψήφιος δείκτης κυτταρικής γήρανσης. Επιπρόσθετα, κατεψυγμένοι ιστοί θετικοί για SA-β-gal μονιμοποιήθηκαν σε φορμόλη, εγκλείστηκαν σε παραφίνη και βάφτηκαν με SBB. Οι αντίστοιχες SA-β-gal θετικές περιοχές στον ιστό βάφτηκαν ειδικά για λιποφουσκίνη με SBB, ενώ οι ιστοί που ήταν αρνητικοί για SA-β-gal βρέθηκαν και αρνητικοί για τη λιποφουσκίνη. Τα ευρήματα αυτά ενισχύουν περαιτέρω την ευαισθησία και την ειδικότητα της SBB χρώσης για την ανάδειξη γηρασμένων κυττάρων σε αρχειακό υλικό. Η τελευταία μοναδική ιδιότητα του SBB μπορεί να αξιοποιηθεί για κλινικοπαθολογικές μελέτες στο ευρέως διαθέσιμο αρχειακό υλικό. Επιπρόσθετα, η εφαρμογή της τεχνικής σε τομές παραφίνης από Ομαλό λειχήνα, Ακανθοκυτταρικό καρκίνωμα σε ινώματα και φυσιολογικό ιστό στόματος, ανέδειξε την παρουσία γηρασμένων ινοβλαστών στις τομές των ινωμάτων και του Ομαλού λειχήνα και την απουσία τους στο φυσιολογικό ιστό και το Ακανθοκυτταρικό καρκίνωμα του στόματος. Τα ευρήματα αυτά συνηγορούν υπερ μιας προστατευτικής δράσης αυτών των κυττάρων , πιθανά στα πλαίσια μιας καλοήθους αντίδρασης του στρώματος.

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Loss of SATB1 Induces a p21 Dependent Cellular Senescence Phenotype in Dopaminergic Neurons

10.1101/452243 ◽

2018 ◽

Cited By ~ 1

Author(s):

Markus Riessland ◽

Benjamin Kolisnyk ◽

Tae Wan Kim ◽

Jia Cheng ◽

Jason Ni ◽

...

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Cellular Senescence ◽

Dopaminergic Neurons ◽

Dna Binding Protein ◽

Dopamine Neurons ◽

Human Stem Cell ◽

Transcriptional Program

AbstractCellular senescence is a mechanism used by mitotic cells to prevent uncontrolled cell division. As senescent cells persist in tissues, they cause local inflammation and are harmful to surrounding cells, contributing to aging. Generally, neurodegenerative diseases, such as Parkinson‘s, are disorders of aging. The contribution of cellular senescence to neurodegeneration is still unclear. SATB1 is a DNA binding protein associated with Parkinson’s disease. We report that SATB1 prevents cellular senescence in post-mitotic dopaminergic neurons. Loss of SATB1 causes activation of a cellular senescence transcriptional program in dopamine neurons, both in human stem cell-derived dopaminergic neurons and in mice. We observed phenotypes which are central to cellular senescence in SATB1 knockout dopamine neurons in vitro and in vivo. Moreover, we found that SATB1 directly represses expression of the pro-senescence factor, p21, in dopaminergic neurons. Our data implicate senescence of dopamine neurons as a contributing factor to the pathology of Parkinson’s disease.

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Exploring the effect of nsSNPs in human YPEL3 gene in cellular senescence

Scientific Reports ◽

10.1038/s41598-020-72333-8 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Abhishek Singh ◽

Mukesh Thakur ◽

Sujeet Kumar Singh ◽

Lalit Kumar Sharma ◽

Kailash Chandra

Keyword(s):

Conformational Changes ◽

Cellular Senescence ◽

Health Professionals ◽

Md Simulation ◽

Tumour Cells ◽

In Vivo Studies ◽

Native Conformation ◽

Altered Expression

Abstract YPEL3 that induces cellular senescence in both normal and tumour cells of humans may show altered expression under the influence of incidental mutations. In this study, we proposed the first structure of Native YPEL3 protein and its five possible deleterious mutants—V40M, C61Y, G98R, G108S, and A131T and predicted their deleterious effects to alter stability, flexibility and conformational changes in the protein. The MD simulation (RMSD, RMSF, Rg, h-bond and SASA) analysis revealed that the variants V40M, G98R and G108S increased the flexibility in protein, and variant V40M imparted more compactness to the protein.. In general, variants attributed changes in the native conformation and structure of the YPEL3 protein which might affect the native function of cellular senescence. The study provides opportunities for health professionals and practitioners in formulating précised medicines to effectively cure various cancers. We propose in-vitro or in-vivo studies should consider these reported nsSNPs while examining any malfunction in the YPEL3 protein.

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P01.16 Effects of the STAT3 inhibitors on senescent tumour cells

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2020-itoc7.29 ◽

2020 ◽

Vol 8 (Suppl 2) ◽

pp. A16.1-A16

Author(s):

O Sapega ◽

R Mikyskova ◽

K Musilek ◽

J Bieblova ◽

Z Hodny ◽

...

Keyword(s):

Cell Proliferation ◽

Czech Republic ◽

Cell Lines ◽

Cellular Senescence ◽

Tumour Cells ◽

Stat3 Signaling ◽

Stat3 Signaling Pathway ◽

Stat3 Phosphorylation

BackgroundCellular senescence is the process of cell proliferation arrest. Premature cellular senescence can be induced by chemotherapy, irradiation and, under certain circumstances, by cytokines. Senescent cells produce a number of secreted proteins and growth factors that may either stimulate or inhibit cell proliferation. One of the major cytokines that play role in regulation of cellular senescence is IL-6. IL-6/STAT3 signaling pathway represent decisive regulatory factors in cellular senescence. The objective of this study was to compare the effects of the STAT3 inhibitors on senescent and proliferative tumour cells. Further, the therapeutic potential of the STAT3 inhibitors was evaluated using murine tumour models.Materials and MethodsIn vitro, as well as in vivo experiments were performed using TC-1 (model for HPV16-associated tumours) TRAMP-C2 (prostate cancer) cell lines. C57Bl/6NCrl mice, 7–8 weeks old, were obtained from Velaz (Prague, Czech Republic). Experimental protocols were approved by the Institutional Animal Care Committee of the Institute of Molecular Genetics (Prague, Czech Republic). STAT3 inhibitors, namely STATTIC, BP-102 (synthesised at the University of Hradec Kralove) and their derivatives were tested for their effects on tumour cells, such as cytotoxicity, ability to inhibit STAT3 phosphorylation, cell proliferation and tumour growth in syngeneic mice.ResultsWe have previously demonstrated that docetaxel-induced senescence in the TC-1 and TRAMP-C2 murine tumour cell lines, which was proved by in vitro (detection of increased p21 expression, positive beta-galactosidase staining, and the typical SASP capable to induce ‘bystander’ senescence), and in vivo experiments, using C57BL/6 mice [1]. Both TC-1 and TRAMP-C2 cells displayed elevated IL-6 secretion and activated STAT3 signaling pathway. Therefore, we tested efficacy of the STAT3 inhibitors on these cell lines. Cytotoxic and STAT3 phosphorylation inhibitory effects of the inhibitors were observed in both proliferating and senescent cells. Antitumor effects of selected inhibitors were evaluated.ConclusionsCollectively, STAT3 is an attractive target for therapeutic approaches in cancer treatment and we can assume that inhibition of the STAT3 pathway can be used for elimination of the pernicious effects of the senescent cells.ReferenceSimova J, Sapega O, Imrichova T, Stepanek I, Kyjacova L, Mikyskova R, Indrova M, Bieblova J, Bubenik J, Bartek J, et al: Tumor growth accelerated by chemotherapy-induced senescent cells is suppressed by treatment with IL-12 producing cellular vaccines. Oncotarget7: 54952–54964, 2016. This work was supported by the research grant No. NV18-05-00562 provided by the Grant Agency of the Ministry of Health of the Czech Republic.Disclosure InformationO. Sapega: None. R. Mikyskova: None. K. Musilek: None. J. Bieblova: None. Z. Hodny: None. M. Reinis: None.

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