Deregulation of focal adhesion formation and cytoskeletal tension due to loss of A-type lamins

Corne, Tobias; Sieprath, Tom; Vandenbussche, Jonathan; Mohammed, Danahé; Lindert, Mariska te; Gevaert, Kris; Gabriele, Sylvain; Wolf, Katarina; Vos, Winnok H. De

doi:10.1080/19336918.2016.1247144

Cited by 24 publications

(18 citation statements)

References 72 publications

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“…To rule out a major effect on the general migration machinery after lamin depletion (Corne et al, 2016;Hale et al, 2008), we tested migration across ECM-coated glass and large pore 3D conditions and detected no principal migration defect when cells move in non-confining conditions. When, however, comparing speed changes under confinement, downregulation of lamins allow acceleration by 1.5-fold in 3D collagen and PDMS channels and up to 2.5-fold crossing transwell membranes.…”

Section: Implications For Cell Navigation In Complex Environmentsmentioning

confidence: 99%

Lamin B2 follows lamin A/C- mediated nuclear mechanics and cancer cell invasion efficacy

Vortmeyer-Krause

Lindert

Riet

et al. 2020

Preprint

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2Abbreviations: 2D, two-dimensional; 3D, three-dimensional; AFS, atomic force spectroscopy; ECM, extracellular matrix; H2B, histone-2B; MMP matrix metalloproteinase; NII, nuclear irregularity index; NT, non-targeting control; PDMS, polydimethylsiloxane SummaryNuclear deformability during cancer cell invasion and metastasis is critically regulated by lamin A. Here, researchers showed that lamin B2 also contributes to nuclear mechanics, implicating cooperating lamin networks regulating nuclear integrity, migration efficacy, and metastatic tumor progression. AbstractInterstitial tumor cell invasion depends upon complex mechanochemical adaptation of both cell body and the rigid nucleus in response to extracellular tissue topologies. Nuclear mechanics during cell migration through confined environments is controlled by A-type lamins, however, the contribution of B-type lamins to the deformability of the nucleus remains unclear. Using systematic expression regulation of different lamin isoforms, we applied multi-parameter wet-lab and in silico analysis to test their impact on nuclear mechanics, shape regulation, and cancer cell migration. Modulation of lamin A/C and B2 but not B1 isoforms controlled nuclear deformation and viscoelasticity, whereby lamin B2 generally followed lamin A/C-mediated effects. Cell migration rates were altered by 5 to 9fold in dense collagen environments and synthetic devices, with accelerated rates after lamin downregulation and reverse effects after lamin upregulation, with migration rates strongly depending on nuclear shape change. These findings implicate cooperation of lamin B2 with lamin A/C in regulating nuclear mechanics for shape adaptation and migration efficacy.

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Section: Implications For Cell Navigation In Complex Environmentsmentioning

confidence: 99%

Lamin B2 follows lamin A/C- mediated nuclear mechanics and cancer cell invasion efficacy

Vortmeyer-Krause

Lindert

Riet

et al. 2020

Preprint

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“…After automatic thresholding (Isodata algorithm), touching nuclei are separated using a conditional watershed algorithm ( 40 ). Next, cytoplasmic ROIs are determined as concentric bands around the nuclei by Voronoi-restrained region growing ( 41 ). Finally, the average intensity of the GR channel is measured in both the nuclear and cytoplasmic ROIs and the nuclear-to cytoplasm signal intensity ratio (N/C Ratio) is calculated per cell (Figure S1 in Supplementary Material) ( 42 ).…”

Section: Methodsmentioning

confidence: 99%

Selective Glucocorticoid Receptor Properties of GSK866 Analogs with Cysteine Reactive Warheads

et al. 2017

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Synthetic glucocorticoids (GC) are the mainstay therapy for treatment of acute and chronic inflammatory disorders. Due to the high adverse effects associated with long-term use, GC pharmacology has focused since the nineties on more selective GC ligand-binding strategies, classified as selective glucocorticoid receptor (GR) agonists (SEGRAs) or selective glucocorticoid receptor modulators (SEGRMs). In the current study, GSK866 analogs with electrophilic covalent-binding warheads were developed with potential SEGRA properties to improve their clinical safety profile for long-lasting topical skin disease applications. Since the off-rate of a covalently binding drug is negligible compared to that of a non-covalent drug, its therapeutic effects can be prolonged and typically, smaller doses of the drug are necessary to reach the same level of therapeutic efficacy, thereby potentially reducing systemic side effects. Different analogs of SEGRA GSK866 coupled to cysteine reactive warheads were characterized for GR potency and selectivity in various biochemical and cellular assays. GR- and NFκB-dependent reporter gene studies show favorable anti-inflammatory properties with reduced GR transactivation of two non-steroidal GSK866 analogs UAMC-1217 and UAMC-1218, whereas UAMC-1158 and UAMC-1159 compounds failed to modulate cellular GR activity. These results were further supported by GR immuno-localization and S211 phospho-GR western analysis, illustrating significant GR phosphoactivation and nuclear translocation upon treatment of GSK866, UAMC-1217, or UAMC-1218, but not in case of UAMC-1158 or UAMC-1159. Furthermore, mass spectrometry analysis of tryptic peptides of recombinant GR ligand-binding domain (LBD) bound to UAMC-1217 or UAMC-1218 confirmed covalent cysteine-dependent GR binding. Finally, molecular dynamics simulations, as well as glucocorticoid receptor ligand-binding domain (GR-LBD) coregulator interaction profiling of the GR-LBD bound to GSK866 or its covalently binding analogs UAMC-1217 or UAMC-1218 revealed subtle conformational differences that might underlie their SEGRA properties. Altogether, GSK866 analogs UAMC-1217 and UAMC-1218 hold promise as a novel class of covalent-binding SEGRA ligands for the treatment of topical inflammatory skin disorders.

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“…both depletion of Lamin A/C[53] and nucleo-cytoskeletal connector Nesprin-1[54] were shown to increase substrate traction in cells. Tracking with increased basal pFAK levels, application of either LIV or strain pushed acute FAK phosphorylation of Lamin A/C depleted cells higher than control cells.…”

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confidence: 93%

Lamin A/C functions independently from mechanical signaling during adipogenesis

Goelzer

Dudakovic

Olcum

et al. 2020

Preprint

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Mesenchymal stem cells (MSC) maintain the musculoskeletal system by differentiating into multiple cell types including osteocytes and adipocytes. Mechanical signals, including strain and low intensity vibration (LIV), are important regulators of MSC differentiation. Lamin A/C is a vital protein for nuclear architecture that supports chromatin organization, as well as mechanical integrity and mechano-sensitivity of the nucleus in MSCs. Here, we investigated whether Lamin A/C and mechano-responsiveness are functionally coupled during adipogenesis. Lamin depletion in MSCs using siRNA increased nuclear area, height and volume and decreased circularity and stiffness, while phosphorylation of focal adhesions and dynamic substrate strain in response to LIV remained intact. Lamin A/C depletion decelerates adipogenesis as reflected by delayed appearance of key biomarkers (e.g., adiponectin/ADIPOQ). Based on RNA-seq data, reduced Lamin A/C levels decrease the activation of the adipocyte transcriptome that is normally observed in response to adipogenic cues mediating differentiation of MSCs. Mechanical stimulation via daily LIV application reduced the expression levels of ADIPOQ in both control and Lamin A/C depleted cells. Yet, treatment with LIV did not induce major transcriptome changes in either control or Lamin A/C depleted MSCs, suggesting that the biological effects of LIV on adipogenesis may not occur at the transcriptional level. We conclude that while Lamin A/C activation is essential for normal adipogenesis, it is dispensible for activation of focal adhesions by dynamic vibration induced mechanical signals.

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Deregulation of focal adhesion formation and cytoskeletal tension due to loss of A-type lamins

Cited by 24 publications

References 72 publications

Lamin B2 follows lamin A/C- mediated nuclear mechanics and cancer cell invasion efficacy

Lamin B2 follows lamin A/C- mediated nuclear mechanics and cancer cell invasion efficacy

Selective Glucocorticoid Receptor Properties of GSK866 Analogs with Cysteine Reactive Warheads

Lamin A/C functions independently from mechanical signaling during adipogenesis

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