Dmitri Gordienko - Academia.edu (original) (raw)

Papers by Dmitri Gordienko

Nature Communications, 2022

Cellular senescence is implicated in a great number of diseases including cancer. Although altera... more Cellular senescence is implicated in a great number of diseases including cancer. Although alterations in mitochondrial metabolism were reported as senescence drivers, the underlying mechanisms remain elusive. We report the mechanism altering mitochondrial function and OXPHOS in stress-induced senescent fibroblasts. We demonstrate that TRPC3 protein, acting as a controller of mitochondrial Ca2+ load via negative regulation of IP3 receptor-mediated Ca2+ release, is down regulated in senescence regardless of the type of senescence inducer. This remodelling promotes cytosolic/mitochondrial Ca2+ oscillations and elevates mitochondrial Ca2+ load, mitochondrial oxygen consumption rate and oxidative phosphorylation. Re-expression of TRPC3 in senescent cells diminishes mitochondrial Ca2+ load and promotes escape from OIS-induced senescence. Cellular senescence evoked by TRPC3 downregulation in stromal cells displays a proinflammatory and tumour-promoting secretome that encourages cancer epi...

Journal of Cell Biology, May 27, 2013

Neurophysiology, May 1, 2003

Two layers of interstitial cells (ICs) of Cajal were detected by c-kit and methylene blue stainin... more Two layers of interstitial cells (ICs) of Cajal were detected by c-kit and methylene blue staining in the media of the rabbit portal vein in subendothelial intramuscular and deeper intramuscular positions, displaced radially from each other by about 40-70 microm. Two morphologically distinct types of ICs were found among enzymatically dispersed cells from this vessel: small multipolar cells with stellate-shaped bodies not exceeding 20 microm, and spindle-shaped cells from 40 to 300 microm in length with numerous branching processes. Relaxed smooth muscle cells (SMCs) had a more constant length (90-150 microm). The cell membrane capacitance was 46.5+/-2.2 pF in SMCs, 39.7+/-2.4 pF in spindle-shaped ICs and 27.8+/-0.7 pF in multipolar ICs. Although darker under phase contrast, after loading with fluo-4 AM, single isolated ICs of both types usually had brighter fluorescence than SMCs and displayed various spontaneous calcium events, including Ca(2+) sparks and Ca(2+) waves. Ca(2+) waves were usually followed by contraction of SMCs but no change in shape of ICs. In some ICs spontaneous [Ca(2+)](i) transients (lasting about 2s) which propagated towards the end of the processes were observed. Physical contacts between the processes of ICs and the body of one or more SMCs survived the isolation procedure. Application of noradrenaline (1-10 microM), caffeine (1-10 mM) or high-K(+) solution (60mM) led to a rise of [Ca(2+)](i) in both SMCs and ICs evoking contraction of SMCs but not ICs. No differences in electrophysiological characteristics between single enzymatically isolated IC and SMC were detected; thus, the resting membrane potential estimated under current-clamp conditions was -46.5+/-2.0 mV in spindle-shaped ICs and -45.6+/-2.7 mV in SMCs. Under voltage-clamp, both ICs and SMCs revealed a well-developed voltage-gated nifedipine-sensitive L-type Ca(2+) current, a set of K(+) currents, including spontaneous transient outward currents (STOCs) but no Na(+) current. This study for the first time directly demonstrated the presence in vascular tissue of ICs. Possible roles for ICs including their involvement in spontaneous activity of the vessel were discussed.

Proceedings of the National Academy of Sciences, 2013

SignificanceHuman adenoviruses encode Early region 3 (E3) proteins that manipulate the host immun... more SignificanceHuman adenoviruses encode Early region 3 (E3) proteins that manipulate the host immune response to establish an infection or to persist longer. To date, only a few E3 functions from a single adenovirus species (C) have been characterized, all of which act directly on infected cells. Here we describe a secreted E3 protein that is uniquely expressed by species D adenoviruses. This protein targets noninfected leukocytes using a cell surface phosphatase as a receptor. We provide evidence that this interaction suppresses leukocyte activation and effector functions, implying that species D adenoviruses can affect the host distant from the site of infection.

Scientific Reports, 2019

In prostate carcinogenesis, expression and/or activation of the Transient Receptor Potential Mela... more In prostate carcinogenesis, expression and/or activation of the Transient Receptor Potential Melastatin 8 channel (TRPM8) was shown to block in vitro Prostate Cancer (PCa) cell migration. Because of their localization at the plasma membrane, ion channels, such as TRPM8 and other membrane receptors, are promising pharmacological targets. The aim of this study was thus to use nanocarriers encapsulating a TRPM8 agonist to efficiently activate the channel and therefore arrest PCa cell migration. To achieve this goal, the most efficient TRPM8 agonist, WS12, was encapsulated into Lipid NanoCapsules (LNC). The effect of the nanocarriers on channel activity and cellular physiological processes, such as cell viability and migration, were evaluated in vitro and in vivo. These results provide a proof-of-concept support for using TRPM8 channel-targeting nanotechnologies based on LNC to develop more effective methods inhibiting PCa cell migration in zebrafish xenograft.

The FASEB Journal, Apr 1, 2009

The EMBO Journal, Jun 6, 2023

There is growing evidence that ion channels are critically involved in cancer cell invasiveness a... more There is growing evidence that ion channels are critically involved in cancer cell invasiveness and metastasis. However, the molecular mechanisms of ion signaling promoting cancer behavior are poorly understood and the complexity of the underlying remodeling during metastasis remains to be explored. Here, using a variety of in vitro and in vivo techniques, we show that metastatic prostate cancer cells acquire a specific Na+/Ca2+ signature required for persistent invasion. We identify the Na+ leak channel, NALCN, which is overexpressed in metastatic prostate cancer, as a major initiator and regulator of Ca2+ oscillations required for invadopodia formation. Indeed, NALCN‐mediated Na+ influx into cancer cells maintains intracellular Ca2+ oscillations via a specific chain of ion transport proteins including plasmalemmal and mitochondrial Na+/Ca2+ exchangers, SERCA and store‐operated channels. This signaling cascade promotes activity of the NACLN‐colocalized proto‐oncogene Src kinase, actin remodeling and secretion of proteolytic enzymes, thus increasing cancer cell invasive potential and metastatic lesions in vivo. Overall, our findings provide new insights into an ion signaling pathway specific for metastatic cells where NALCN acts as persistent invasion controller.

Pharmacological Reports, Jun 1, 2014

Background: ATP is one of the principal sympathetic neurotransmitters which contracts vascular sm... more Background: ATP is one of the principal sympathetic neurotransmitters which contracts vascular smooth muscle cells (SMCs) via activation of ionotropic P2X receptors (P2XRs). We have recently demonstrated that contraction of the guinea pig small mesenteric arteries evoked by stimulation of P2XRs is sensitive to inhibitors of IP 3 receptors (IP 3 Rs). Here we analyzed contribution of IP 3 Rs and ryanodine receptors (RyRs) to [Ca 2+ ] i transients induced by P2XR agonist ab-meATP (10 mM) in single SMCs from these vessels. Methods: The effects of inhibition of L-type Ca 2+ channels (VGCCs), RyRs and IP 3 Rs (5 mM nicardipine, 100 mM tetracaine and 30 mM 2-APB, respectively) on ab-meATP-induced [Ca 2+ ] i transients were analyzed using fast x-y confocal Ca 2+ imaging. Results: The effect of IP 3 R inhibition on the [Ca 2+ ] i transient was significantly stronger (67 AE 7%) than that of RyR inhibition (40 AE 5%) and was attenuated by block of VGCCs. The latter indicates that activation of VGCCs is linked to IP 3 R-mediated Ca 2+ release. Immunostaining of RyRs and IP 3 Rs revealed that RyRs are located mainly in deeper sarcoplasmic reticulum (SR) while sub-plasma membrane (PM) SR elements are enriched with type 1 IP 3 Rs. This structural peculiarity makes IP 3 Rs more accessible to Ca 2+ entering the cell via VGCCs. Thus, IP 3 Rs may serve as an ''intermediate amplifier'' between voltage-gated Ca 2+ entry and RyRmediated Ca 2+ release. Conclusions: P2X receptor activation in mesenteric artery SMCs recruits IP 3 Rs-mediated Ca 2+ release from sub-PM SR, which is facilitated by activation of VGCCs. Sensitivity of IP 3 R-mediated release to VGCC antagonists in vascular SMCs makes this mechanism of special therapeutic significance.

Scientific Reports, May 28, 2019

The Journal of Physiology, Aug 1, 2002

In smooth muscle cells freshly isolated from rabbit portal vein, there was only one site discharg... more In smooth muscle cells freshly isolated from rabbit portal vein, there was only one site discharging the majority of spontaneous Ca2+-release events; the activity of this single site was studied using laser scanning confocal imaging after loading the cells with the fluorescent Ca2+ indicator fluo-4 acetoxymethyl ester. Localised spontaneous Ca2+-release events visualised by line-scan imaging revealed two predominant spatiotemporal patterns: (i) small-amplitude, fast events similar to Ca2+ sparks in cardiomyocytes and (ii) larger and slower events. The sum of two Gaussian profiles was well fitted to the amplitude histogram (peak frequencies at 1.8 and 3.2 F/F0) and spatial spread (full width at half-maximal amplitude) histogram (peak frequencies at 2 and 3.8 μm) for the 230 localised Ca2+-release events analysed. The existence of two populations of Ca2+-release events was also supported by the histograms of the rise times and half-decay times, which revealed modes at 38 and 65 ms, respectively. Shifting the scan line along the z-axis during imaging from a single discharge site suggested that the appearance of two populations of Ca2+-release events is not due to out-of-focus imaging. Both small and large events persisted upon 3–5 min exposure to 1–5 μm nicardipine, but were abolished after 10–15 min exposure to 50–100 μm ryanodine, 0.1 μm thapsigargin or 10 μm cyclopiazonic acid. Only small-amplitude, fast events persisted in the presence of inhibitors of inositol 1,4,5-trisphosphate (IP3)-induced Ca2+ release, 10 μm xestospongin C or 30 μm 2-aminoethoxy-diphenylborate (2-APB), or in the presence of 2.5 μm U-73122 (a phospholipase C (PLC) inhibitor). Coupling between neighbouring Ca2+-release domains giving rise to spontaneous [Ca2+]i waves was abolished in the presence of 2-APB. Examination of the saltatory propagation of the waves suggested that the critical factor that determines propagation between domains is a time-dependent change in the sensitivity of ryanodine receptors and/or IP3 receptors to Ca2+, which can give rise to ‘loose coupling’ between release sites. These results suggest that activation of IP3 receptors (due to the tonic activity of PLC and ongoing production of IP3) recruits neighbouring domains of ryanodine receptors, leading to larger Ca2+ releases and saltatory propagation of [Ca2+]i waves in portal vein myocytes.

Journal of Cell Biology, Aug 14, 2006

A lthough human pannexins (PanX) are homologous to gap junction molecules, their physiological fu... more A lthough human pannexins (PanX) are homologous to gap junction molecules, their physiological function in vertebrates remains poorly understood. Our results demonstrate that overexpression of PanX1 results in the formation of Ca 2+-permeable gap junction channels between adjacent cells, thus, allowing direct intercellular Ca 2+ diffusion and facilitating inter cellular Ca 2+ wave propagation. More intriguingly, our results strongly suggest that PanX1 may also form Ca 2+-permeable channels in the endoplasmic reticulum (ER). These channels contribute to the ER Ca 2+ leak and thereby affect the ER Ca 2+ load. Because leakage remains the most enigmatic of those processes involved in intracellular calcium homeostasis, and the molecular nature of the leak channels is as yet unknown, the results of this work provide new insight into calcium signaling mechanisms. These results imply that for vertebrates, a new protein family, referred to as pannexins, may not simply duplicate the connexin function but may also provide additional pathways for intra-and intercellular calcium signaling and homeostasis.

American Journal of Physiology-lung Cellular and Molecular Physiology, Mar 1, 2009

Mitochondria are proposed to be a major oxygen sensor in hypoxic pulmonary vasoconstriction (HPV)... more Mitochondria are proposed to be a major oxygen sensor in hypoxic pulmonary vasoconstriction (HPV), a unique response of the pulmonary circulation to low oxygen tension. Mitochondrial factors including reactive oxygen species, cytochrome c, ATP, and magnesium are potent modulators of voltage-gated K ϩ (Kv) channels in the plasmalemmal membrane of pulmonary arterial (PA) smooth muscle cells (PASMCs). Mitochondria have also been found close to the plasmalemmal membrane in rabbit main PA smooth muscle sections. Therefore, we hypothesized that differences in mitochondria localization in rat PASMCs and systemic mesenteric arterial smooth muscle cells (MASMCs) may contribute to the divergent oxygen sensitivity in the two different circulations. Cellular localization of mitochondria was compared with immunofluorescent labeling, and differences in functional coupling between mitochondria and Kv channels was evaluated with the patch-clamp technique and specific mitochondrial inhibitors antimycin A (acting at complex III of the mitochondrial electron transport chain) and oligomycin A (which inhibits the ATP synthase). It was found that mitochondria were located significantly closer to the plasmalemmal membrane in PASMCs compared with MASMCs. Consistent with these findings, the effects of the mitochondrial inhibitors on Kv current (IKv) were significantly more potent in PASMCs than in MASMCs. The cytoskeletal disruptor cytochalasin B (10 M) also altered mitochondrial distribution in PASMCs and significantly attenuated the effect of antimycin A on the voltage-dependent parameters of IKv. These findings suggest a greater structural and functional coupling between mitochondria and Kv channels specifically in PASMCs, which could contribute to the regulation of PA excitability in HPV. pulmonary artery; vascular smooth muscle cells; mesenteric artery; K ϩ channel activation; K ϩ channel inactivation; confocal imaging; patch-clamp technique

British Journal of Pharmacology, 2004

In visceral smooth muscles, both M 2 and M 3 muscarinic receptor subtypes are found, and produce ... more In visceral smooth muscles, both M 2 and M 3 muscarinic receptor subtypes are found, and produce two major metabolic effects: adenylyl cyclase inhibition and PLCb activation. Thus, we studied their relevance for muscarinic cationic current (mI CAT) generation, which underlies cholinergic excitation. Experiments were performed on single guinea-pig ileal cells using patch-clamp recording techniques under conditions of weakly buffered [Ca 2 þ ] i (either using 50 mM EGTA or 50-100 mM fluo-3 for confocal fluorescence imaging) or with [Ca 2 þ ] i 'clamped' at 100 nM using 10 mM BAPTA/CaCl 2 mixture. 2 Using a cAMP-elevating agent (1 mM isoproterenol) or a membrane-permeable cAMP analog (10 mM 8-Br-cAMP), we found no evidence for mI CAT modulation through a cAMP/PKA pathway. 3 With low [Ca 2 þ ] i buffering, the PLC blocker U-73122 at 2.5 mM almost abolished mI CAT , in some cases without any significant effect on [Ca 2 þ ] i. When [Ca 2 þ ] i was buffered at 100 nM, U-73122 reduced both carbachol-and GTPgS-induced mI CAT maximal conductances (IC 50 ¼ 0.5-0.6 mM) and shifted their activation curves positively. 4 U-73343, a weak PLC blocker, had no effect on GTPgS-induced mI CAT , but weakly inhibited carbachol-induced current, possibly by competitively inhibiting muscarinic receptors, since the inhibition could be prevented by increasing the carbachol concentration to 1 mM. Aristolochic acid and D-609, which inhibit PLA 2 and phosphatidylcholine-specific PLC, respectively, had no or very small effects on mI CAT , suggesting that these enzymes were not involved. 5 InsP 3 (1 mM) in the pipette or OAG (20 mM) applied externally had no effect on mI CAT or its inhibition by U-73122. Ca 2 þ store depletion (evoked by InsP 3 , or by combined cyclopiazonic acid, ryanodine and caffeine treatment) did not induce any significant current, and had no effect on mI CAT in response to carbachol when [Ca 2 þ ] i was strongly buffered to 100 nM. 6 It is concluded that phosphatidylinositol-specific PLC modulates mI CAT via Ca 2 þ release, but also does so independently of InsP 3 , DAG, Ca 2 þ store depletion or a rise of [Ca 2 þ ] i. Our present results explain the previously established 'permissive' role of the M 3 receptor subtype in mI CAT generation, and provide a new insight into the molecular mechanisms underlying the shifts of the cationic conductance activation curve.

Cell Calcium, Apr 1, 2003

Nature Communications, Feb 17, 2022

Cellular senescence is implicated in a great number of diseases including cancer. Although altera... more Cellular senescence is implicated in a great number of diseases including cancer. Although alterations in mitochondrial metabolism were reported as senescence drivers, the underlying mechanisms remain elusive. We report the mechanism altering mitochondrial function and OXPHOS in stress-induced senescent fibroblasts. We demonstrate that TRPC3 protein, acting as a controller of mitochondrial Ca 2+ load via negative regulation of IP 3 receptormediated Ca 2+ release, is down regulated in senescence regardless of the type of senescence inducer. This remodelling promotes cytosolic/mitochondrial Ca 2+ oscillations and elevates mitochondrial Ca 2+ load, mitochondrial oxygen consumption rate and oxidative phosphorylation. Re-expression of TRPC3 in senescent cells diminishes mitochondrial Ca 2+ load and promotes escape from OIS-induced senescence. Cellular senescence evoked by TRPC3 downregulation in stromal cells displays a proinflammatory and tumour-promoting secretome that encourages cancer epithelial cell proliferation and tumour growth in vivo. Altogether, our results unravel the mechanism contributing to pro-tumour behaviour of senescent cells.

Although human pannexins (PanX) are homologous to gap junction molecules, their physiological fun... more Although human pannexins (PanX) are homologous to gap junction molecules, their physiological function in vertebrates remains poorly understood. Our results demonstrate that overexpression of PanX1 results in the formation of Ca 2+-permeable gap junction channels between adjacent cells, thus, allowing direct intercellular Ca 2+ diffusion and facilitating inter cellular Ca 2+ wave propagation. More intriguingly, our results strongly suggest that PanX1 may also form Ca 2+-permeable channels in the endoplasmic reticulum (ER). These channels contribute to the ER Ca 2+ leak and

International Review of Cell and Molecular Biology, 2021

In recent decades cancer emerged as one of the leading causes of death in the developed countries... more In recent decades cancer emerged as one of the leading causes of death in the developed countries, with some types of cancer contributing to the top 10 causes of death on the list of the World Health Organization. Carcinogenesis, a malignant transformation causing formation of tumors in normal tissues, is associated with changes in the cell cycle caused by suppression of signaling pathways leading to cell death and facilitation of those enhancing proliferation. Further progression of cancer, during which benign tumors acquire more aggressive phenotypes, is characterized by metastatic dissemination through the body driven by augmented motility and invasiveness of cancer cells. All these processes are associated with alterations in calcium homeostasis in cancer cells, which promote their proliferation, motility and invasion, and dissuade cell death or cell cycle arrest. Remodeling of store-operated calcium entry (SOCE), one of the major pathways regulating intracellular Ca2+ concentration ([Ca2+]i), manifests a key event in many of these processes. This review systematizes current knowledge on the mechanisms recruiting SOCE-related proteins in carcinogenesis and cancer progression.

The FASEB Journal, 2020

Complex wrinkle patterns have been observed in various thin film systems, typically with integrat... more Complex wrinkle patterns have been observed in various thin film systems, typically with integrated hard and soft materials for various applications as well as in nature. The underlying mechanism of wrinkling has been generally understood as a stress-driven instability. On an elastic substrate, equilibrium and energetics set the critical condition and select the wrinkle wavelength and amplitude. On a viscous substrate, wrinkles grow over time and kinetics select the fastest growing mode. Moreover, on a viscoelastic substrate, both energetics and kinetics play important roles in determining the critical condition, the growth rate, and wrinkle patterns. The dynamics of wrinkling, while analogous to other phase ordering phenomena, is rich and distinct under the effects of stress and film-substrate interactions. In this chapter, a kinetics approach is presented for wrinkling of isotropic and anisotropic elastic films on viscoelastic substrates. Analytic solutions are obtained by a linear perturbation analysis and a nonlinear energy minimization method, which predict the kinetics of wrinkle growth at the early stage and the equilibrium states at the long-time limit, respectively. In between, a power-law coarsening of the wrinkle wavelength is predicted by a scaling analysis. Furthermore, the kinetics approach enables numerical simulations that demonstrate emergence and transition of diverse wrinkle patterns (ordered and disordered) under various conditions.

Cytosolic Ca2+ oscillations provide signaling input to several effector systems of the cell. Thes... more Cytosolic Ca2+ oscillations provide signaling input to several effector systems of the cell. These include neuronal development, migration and networking. Although similar signaling events are hijacked by highly aggressive cancer cells, the complexity of the ‘neuron-like’ remodeling in metastasis remains to be explored. Here, using a variety of in vitro and in vivo techniques we show that strongly metastatic prostate cancer cells acquire specific Na+/Ca2+ signature required for persistent invasion. We identify the ‘neuronal’ Na+ leak channel, NALCN, at the hot spots of the Ca2+ wave initiation and invadopodia formation. Mechanistically, NALCN associates functionally with plasmalemmal and mitochondrial Na+/Ca2+ exchangers, reactive oxygen species and store-operated channels to generate intracellular Ca2+ oscillations. In turn, this stimulates the activity of protooncogene Src kinase co-localized with NALCN, actin remodeling and secretion of proteolytic enzymes, thus increasing an inv...

Nature Communications, 2022

Journal of Cell Biology, May 27, 2013

Neurophysiology, May 1, 2003

Proceedings of the National Academy of Sciences, 2013

Scientific Reports, 2019

The FASEB Journal, Apr 1, 2009

The EMBO Journal, Jun 6, 2023

Pharmacological Reports, Jun 1, 2014

Scientific Reports, May 28, 2019

The Journal of Physiology, Aug 1, 2002

Journal of Cell Biology, Aug 14, 2006

American Journal of Physiology-lung Cellular and Molecular Physiology, Mar 1, 2009

British Journal of Pharmacology, 2004

Cell Calcium, Apr 1, 2003

Nature Communications, Feb 17, 2022

Cellular senescence is implicated in a great number of diseases including cancer. Although altera... more Cellular senescence is implicated in a great number of diseases including cancer. Although alterations in mitochondrial metabolism were reported as senescence drivers, the underlying mechanisms remain elusive. We report the mechanism altering mitochondrial function and OXPHOS in stress-induced senescent fibroblasts. We demonstrate that TRPC3 protein, acting as a controller of mitochondrial Ca 2+ load via negative regulation of IP 3 receptormediated Ca 2+ release, is down regulated in senescence regardless of the type of senescence inducer. This remodelling promotes cytosolic/mitochondrial Ca 2+ oscillations and elevates mitochondrial Ca 2+ load, mitochondrial oxygen consumption rate and oxidative phosphorylation. Re-expression of TRPC3 in senescent cells diminishes mitochondrial Ca 2+ load and promotes escape from OIS-induced senescence. Cellular senescence evoked by TRPC3 downregulation in stromal cells displays a proinflammatory and tumour-promoting secretome that encourages cancer epithelial cell proliferation and tumour growth in vivo. Altogether, our results unravel the mechanism contributing to pro-tumour behaviour of senescent cells.

International Review of Cell and Molecular Biology, 2021

The FASEB Journal, 2020