Egor Zindy's research while affiliated with The University of Manchester and other places

Publications (47)

Article
Full-text available
Apoptosis is regulated by interactions between the BH3-only and multi-domain Bcl-2 family proteins. These interactions are integrated on the outer mitochondrial membrane (OMM) where they set the threshold for apoptosis, known as mitochondrial priming. However, how mitochondrial priming is controlled at the level of single cells remains unclear. Ret...
Article
Full-text available
Saliva secretion requires effective translocation of aquaporin 5 (AQP5) water channel to the salivary glands (SGs) acinar apical membrane. Patients with Sjögren’s syndrome (SS) display abnormal AQP5 localization within acinar cells from SGs that correlate with sicca manifestation and glands hypofunction. Several proteins such as Prolactin-inducible...
Article
Full-text available
Integration of signalling downstream of individual receptor tyrosine kinases (RTKs) is crucial to fine-tune cellular homeostasis during development and in pathological conditions, including breast cancer. However, how signalling integration is regulated and whether the endocytic fate of single receptors controls such signalling integration remains...
Article
Full-text available
Sjögren’s syndrome (SS) is an exocrinopathy characterized by the hypofunction of salivary glands (SGs). Aquaporin-5 (AQP5); a water channel involved in saliva formation; is aberrantly distributed in SS SG acini and contributes to glandular dysfunction. We aimed to investigate the role of ezrin in AQP5 mislocalization in SS SGs. The AQP5–ezrin inter...
Preprint
Integration of signaling downstream from individual Receptor Tyrosine Kinases (RTKs) is crucial to fine tune cellular homeostasis during development and in pathological conditions, including breast cancer. However, how signalling integration is regulated and whether the endocytic fate of single receptors controls such signalling integration still r...
Preprint
Microenvironmental stiffness regulates the behaviour of both normal and cancer cells. In breast tissue, high mammographic density (HMD), which reflects greater organisation and stiffness of the periductal collagen, represents a significant risk factor for cancer. However, the mechanistic link between extracellular matrix (ECM) stiffness and increas...
Article
Full-text available
Glucocorticoids (GCs) act through the glucocorticoid receptor (GR, also known as NR3C1) to regulate immunity, energy metabolism and tissue repair. Upon ligand binding, activated GR mediates cellular effects by regulating gene expression, but some GR effects can occur rapidly without new transcription. Here, we show that GCs rapidly inhibit cell mig...
Article
Full-text available
Evolutionarily conserved circadian clocks generate 24-hour rhythms in physiology and behaviour that adapt organisms to their daily and seasonal environments. In mammals, the suprachiasmatic nucleus (SCN) of the hypothalamus is the principal co-ordinator of the cell-autonomous clocks distributed across all major tissues. The importance of robust dai...
Article
Full-text available
Background: Glioblastoma (GBM) has been extensively researched over the last few decades, yet despite aggressive multi-modal treatment, recurrence is inevitable and second-line treatment options are limited. Here, we demonstrate how high throughput screening (HTS) in multicellular spheroids can generate physiologically relevant patient chemosensit...
Chapter
Small proteinaceous oligomeric species contribute to the formation of larger aggregates, a phenomenon that is of direct relevance to the characterization of protein aggregation in biopharmaceuticals and understanding the underlying processes contributing to neurodegenerative diseases.The ability to monitor in situ oligomerization and aggregation pr...
Article
Full-text available
Background Circadian rhythms maintain tissue homeostasis during the 24-h day-night cycle. Cell-autonomous circadian clocks play fundamental roles in cell division, DNA damage responses and metabolism. Circadian disruptions have been proposed as a contributing factor for cancer initiation and progression, although definitive evidence for altered mol...
Article
The formation of uniaxial fibrous tissues with defined viscoelastic properties implies the existence of an orchestrated mechanical interaction between the cytoskeleton and the extracellular matrix. This study addresses the nature of this interaction. The hypothesis is that this mechanical interplay underpins the mechanical development of the tissue...
Article
Full-text available
The circadian clock is an autonomous molecular feedback loop inside almost every cell in the body. We have shown that the mammary epithelial circadian clock is regulated by the cellular microenvironment. Moreover, a stiff extracellular matrix dampens the oscillations of the epithelial molecular clock. Here, we extend this analysis to other tissues...
Data
Method for calculating average FRET ratio in the leading edge of migrating cells. A. Typical raw YFP-donor YFP-acceptor images at 4 different time points as of a migrating cell on a CDM nucleofected with the Raichu-RhoA probe (as well as Eps8 siRNA and treated with cRGDfV and PD184352). Images were taken with 400ms exposure. B. The same timepoints...
Article
Full-text available
Cell migration in 3D microenvironments is fundamental to development, homeostasis and the pathobiology of diseases such as cancer. Rab-coupling protein (RCP) dependent co-trafficking of α5β1 and EGFR1 promotes cancer cell invasion into fibronectin (FN) containing extracellular matrix (ECM), by potentiating EGFR1 signalling at the front of invasive...
Data
MEK1/2 inhibition effect on Rac1 and RhoA activity of migrating H1299 cells expressing mutant p53. Representative Ratiometric FRET images of whole H1299-mutant p53 expressing cells on CDMs at a single timepoint. A. Cell transfected with Raichu-Rac1 probe, stimulated with cRGDfV and treated with DMSO for vehicle; B. Cell transfected with Raichu-Rac1...
Data
Full list of references for information found in S1 and S2 Tables. (DOCX)
Data
Initially active nodes in simulations and justification. All other nodes in the model are set to OFF and may become active at some time following EGF input. (DOCX)
Data
Contains all required code and instructions to simulate the model in CellNetAnalyzer (the MatLAB plugin). Raw code for reactions and species in the model can also be read by opening the ‘reactions’ or ‘metabolites’ file. (ZIP)
Data
FRET biosensors reporting Rac1 activity (top) or RhoA activity (bottom) for A2780 cells migrating over a 5 minute timecourse on 3D CDMs, stimulated with cRGDfV, treated with MEK1/2 inhibitor PD184352 and transfected with control siRNA (left) or Eps8 siRNA (right) to abrogate formation of the Sos1-Eps8-Abi1 complex. Thermal look up table applied as...
Data
Effects of MEK1/2 inhibition on 2-D A2780 cell migration in scratch wound experiments. A. Representative images of A2780 cells in a sub-domain of an image at t = 0 (the initial frame) and the same sub-domain at t = 5 hours (30 10 minute frames later), for cells without/with cRGDfV stimulation and treated with DMSO or MEK1/2 inhibitor AZD6244. Yello...
Data
Random asynchronous model updates show similar output dynamics. A. Average RhoA and Rac1 node ON time following initial Rac1 OFF and RhoA ON switch during the first 2000 time increments: Asynchronous simulation data taken from 10 simulation repeats, Synchronous simulation data corresponds to deterministic heatmap in Fig 1C and has been scaled for d...
Data
Every interaction in the model and references of origin. (DOCX)
Data
Effect of MEK1/2 inhibition and Eps8 knockdown on 2-D cell migration in scratch wound experiments. A. Representative images of A2780 cells in a sub-domain of an image at t = 0 (the initial frame) and the same sub-domain at t = 5 hours (30 10 minute frames later), for cells without/with cRGDfV stimulation, nucleofected with control siRNA or Eps8 siR...
Data
H1299 cells migrating in a 2D scratch wound assay over > 10 hours, expressing mutant p53, treated with MEK1/2 inhibitor and transfected with control siRNA (left) or Eps8 siRNA (right) to abrogate formation of the Sos1-Eps8-Abi1 complex. (AVI)
Data
Individual Eps8 siRNA renders cells insensitive to MEK1/2 inhibition. A. Average speed and persistence of migrating A2780 cells into a scratch wound, treated exactly as in Fig 4C–4G, with either control siRNA, individual Eps8-A siRNA or individual Eps8-B siRNA as indicated. 3 experimental repeats were performed for each Eps8 siRNA experiment with >...
Data
Full list of Rac1 activator/inhibitor logical hierarchies. (DOCX)
Data
Eps8 siRNA has no direct effect on phosphorylated MAP kinase levels or MEK1/2 inhibition viability. A. Western blots showing total Eps8 levels, endogenous levels of phosphorylated p44 and p42 MAP Kinase (Erk1 and Erk2), and total levels of Akt2 (for loading) for A2780 cells either nucleofected with control siRNA or Eps8 siRNA and treated with PD184...
Data
H1299 cells migrating in a 2D scratch wound assay over > 16 hours, stably transfected with an empty vector for mutant p53 (left) or expressing mutant p53 (right) and treated with DMSO (top) or MEK1/2 inhibitor AZD6244 (bottom). (AVI)
Data
FRET biosensors reporting Rac1 activity (top) or RhoA activity (bottom) for A2780 cells migrating over a 5 minute timecourse on 3D CDMs, stimulated with cRGDfV and treated with DMSO (left) or MEK1/2 inhibitor PD184352 (right). Thermal look up table applied as in Fig 3E–3H, where red denotes high GTPase activity and green denotes low GTPase activity...
Article
Full-text available
The mechanical properties of the cell nucleus change to allow cells to migrate, but how chromatin modifications contribute to nuclear deformability has not been defined. Here, we demonstrate that a major factor in this process involves epigenetic changes that underpin nuclear structure. We investigated the link between cell adhesion and epigenetic...
Article
Full-text available
There is significant evidence that brain-infiltrating CD8+ T cells play a central role in the development of experimental cerebral malaria (ECM) during Plasmodium berghei ANKA infection of C57BL/6 mice. However, the mechanisms through which they mediate their pathogenic activity during malaria infection remain poorly understood. Utilizing intravita...
Article
Invasive migration in 3D extracellular matrix (ECM) is crucial to cancer metastasis, yet little is known of the molecular mechanisms that drive reorganization of the cytoskeleton as cancer cells disseminate in vivo. 2D Rac-driven lamellipodial migration is well understood, but how these features apply to 3D migration is not clear. We find that lame...
Article
Recently, the multifunctional peptide TatLK15 resulting from the fusion of the cell penetrating peptide Tat and the amphipathic peptide LK15 was shown to be efficient at mediating siRNA and shRNA delivery in leukemia cells to silence the bcr-abl oncoprotein. The present study focused on TatLK15 peptide cellular uptake and defining conditions for it...
Article
Full-text available
Real-time confocal imaging was utilised to monitor the in situ loss of BSA monomers and aggregate formation using Spatial Intensity Distribution Analysis (SpIDA) and Raster Image Correlation Spectroscopy (RICS). At the proof of concept level this work has demonstrated the applicability of RICS and SpIDA for monitoring protein oligomerisation and la...
Article
Full-text available
The application of raster image correlation spectroscopy (RICS) as a tool for the characterisation of protein diffusion was assessed using a model protein, bovine serum albumin (BSA), as a function of formulation and denaturing conditions. RICS results were also validated against dynamic light scattering and fluorescence correlation spectroscopy. R...

Citations

... Finally, immunohistochemistry confirmed that PIP staining was faint in the mucus acini, while real-time PCR showed that PIP mRNA was significantly lower in pSS patients when compared to both no-SS sicca subjects and healthy controls, thus supporting the hypothesis that the observed reduction of PIP in pSS saliva may be related to a decrease in that protein's production [25]. Mucin 5 B is a major contributor to the lubricating and viscoelastic properties of whole saliva, while lipocalin plays a role in taste reception, and S-type cystatins have antibacterial and antiviral activity, which is consistent with a protective function [26][27][28]. Analogously, we found that lactoferrin, a globular protein with antibacterial activity, was significantly upregulated in both established and pre-clinical SS [29]. Overall, these proteins may not only play a role in the early diagnosis of pSS but also represent ideal targets for novel therapeutic interventions aimed at restoring salivary function and rheology when curative interventions can, ideally, still be possible. ...
... For example, an interaction between FGFR1 and EGFR can marginally increase epidermal growth factor (EGF)-mediated AKT and STAT3 signaling outputs in lung cancer cells [261]. Along this line, we have recently showed that FGFR2-IgIIIb and EGFR engage in reciprocal regulation of each other's signaling and trafficking in breast cancer cells [275], confirming that FGFR trafficking regulation may become a target for therapeutic intervention. ...
... Broadly, these genes were enriched for roles in managing reactive oxygen species, which are known to be affected by HDAC inhibition [48][49][50] (Supplementary Fig. 14). Unresponsive genes also included G-protein coupled receptors associated with cytoskeleton reorganization, concordant with previous reports of HDAC inhibition preventing glucocorticoid-induced hypertension 51 and changes in microtubule dynamics 52 . Interestingly, we observed various classes of effects on these unresponsive genes suggesting multiple ways in which HDACi might interfere with the DEX response ( Supplementary Fig. 15). ...
... BMAL1 and CLOCK are positive regulators in the circadian rhythm system. BMAL1, a gene essential for the generation of 24-h periodic behavior, is predominantly located in the nucleus, where it encodes proteins with distinct circadian oscillations (Ray et al., 2020;Yang et al., 2020). A two-branch regulation model of circadian clock genes has been proposed, in which the BMAL1-centered circadian clock regulatory network in the body is divided into an autonomous response branch and a memory branch. ...
... Promising drug candidates identified based on such approaches include actinomycin-D. Taylor et al. identified actinomycin-D as a promising candidate among the approved oncology drug set VIII against recurrent GBM, showing that actinomycin-D is more effective than TMZ [145]. Recently, our group also identified actinomycin-D as a highly active compound in IDHmut high-grade glioma cell cultures [146]. ...
... SpIDA analysis was employed to measure the QB and surface density of GPCR proteins in live and fixed cells. GPCRs or associated signaling proteins measured using the SpIDA approach include GABA(B) in spinal cord tissue, tyrosine kinase, secretin receptor, protein aggregation, and kinetics of fluorophore uptake in tissue culture cells [156][157][158][159][160][161]. ...
... The products of clock genes are involved in the expression of p21, Cyc D, c-Myc, and Wee1 for the regulation of the cell cycle [90][91][92]. The period 2 (PER2) has been reported to be a tumor suppressor gene whose expression reduces the formation of a variety of tumors like breast, prostate, and lymphoma [89,93]. Loss of PER2 may induce breast cancer. ...
... (ii) The subsequent relaxation to the steady-state plateau phase reflects Ca 2+dependent channel inactivation, internalization, interactions with modulatory sites within N-and C-termini (16) or negative regulation of Ca 2+ influx by TRPM4 (44). (iii) The relaxation phase was associated with Ca 2+ fluctuations in a frequency range similar to signals reported in glaucomatous TM (45) and pressure-, stretch-and GSK101-stimulated fibroblasts and chondrocytes (49,50). ...
... While several previous studies have characterized the chondrocyte circadian clock and its influence on joint homeostasis, it is not known how the chondrocytes maintain their circadian clock within articular cartilage. It is hypothesized that changes in body temperature and factors within the synovial fluid help with clock synchronization (1,9), but the prolonged maintenance of the circadian clock ex vivo without any synchronizing cues highlights that there might be a factor secreted by the chondrocytes themselves that maintains this synchrony, or the abundant ECM maintains the biochemical and biomechanical niche for synchrony (53). Future work looking into potential synchronizing factors will be key in uncovering this mechanism. ...