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Bogomoletz Institute of Physiology
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Role of ion channels and calcium signaling in malignant transformation and carcinogenesis

This direction of the research is pursued in close collaboration with the Laboratoire de Physiologie Cellulaire, Inserm U1003, Universite de Lille1 (France) headed by Prof. Natalia Prevarskaya. Studies within this direction were supported by two grants from International Association for the Promotion of Co-operation with Scientists from the New Independent States of the Former Soviet Union (INTAS) – INTAS-99-01248 (2000-2003) and INTAS-05-1000008-8223 (2006-2009), and now are continued on initiative basis.

Due to the ability to transport ions through a cell membrane, the activation of various types of ion channels can significantly contribute to the regulation of cell volume, and in the event when they transport calcium ions, also to the changes of intracellular calcium concentration, which plays an important signaling role. The same factors, namely the regulation of cell volume and the changes in calcium signaling, affect the progression of processes that determine cell fate, such as proliferation, differentiation, migration and apoptosis. Since the deregulation of these processes towards enhanced proliferation, loss of differentiation, accelerated migration and increased resistance to apoptosis are characteristic features (also referred to as cancer hallmarks) of malignant transformation of cells, the function of ion channels becomes key to either promotion or prevention of the development of these features. In our studies, the properties and carcinogenic potential of several types of chloride- and calcium-permeable ion channels in prostate cancer cells were described, and suggestions were made regarding their potential as therapeutic targets for cancer treatment. The results of these studies are widely presented in the scientific literature (see relevant publications).

References ►

  1. Reznikov AG, Salivonyk OA, Sotkis AG, Shuba YM. Assessment of gold nanoparticle effect on prostate cancer LNCaP cells. Exp Oncol. 2015 Jun;37(2):100-4.
  2. Prevarskaya N, Skryma R, Shuba Y. Ion channels and the hallmarks of cancer. Trends Mol Med. 2010 Mar;16(3):107-21. Review.
  3. Shuba IaM. Calcium signaling in carcinogenesis. Fiziol Zh. 2007;53(4):110-28. Review. Ukrainian.
  4. Sotkis HV, Lazarenko PM, Boldyriev OI, Voĭtychuk OI, Dosenko VIe, Tumanovs'ka LV, Shuba IaM. Identification of E-4031-sensitive potassium current component in murine P19 embryonic carcinoma cell line differentiated in cardiomyocytes. Fiziol Zh. 2006;52(1):49-61. Ukrainian.
  5. Lemonnier L, Lazarenko R, Shuba Y, Thebault S, Roudbaraki M, Lepage G, Prevarskaya N, Skryma R. Alterations in the regulatory volume decrease (RVD) and swelling-activated Cl- current associated with neuroendocrine differentiation of prostate cancer epithelial cells. Endocr Relat Cancer. 2005 Jun;12(2):335-49.
  6. Lazarenko RM, Kondrats'kyĭ AP, Pohoriela NKh, Shuba IaM. Alterations in ATP-dependence of swelling-activated Cl- current associated with neuroendocrine differentiation of LNCaP human prostate cancer epithelial cells. Fiziol Zh. 2005;51(3):57-66. Ukrainian.
  7. Lazarenko PM, Pohoriela NKh, Shuba IaM. Adenosine triphosphate-dependence of volume sensitive chloride current in LNCaP cell line of human prostate cancer. Fiziol Zh. 2005;51(1):51-61. Ukrainian.
  8. Lazarenko RM, Vitko IuM, Pohoriela NKh, Skryma RN, Shuba IaM. Effect of neuroendocrine cell differentiation of the human prostatic neoplasm cell line LNCaP on the characteristics of volume-sensitive chloride content. Fiziol Zh. 2003;49(6):3-13. Ukrainian.
  9. Vitko YV, Pogorelaya NH, Prevarskaya N, Skryma R, Shuba YM. Proteolytic modification of swelling-activated Cl- current in LNCaP prostate cancer epithelial cells. J Bioenerg Biomembr. 2002 Aug;34(4):307-15.
  10. Vitko IuM, Pogoriela NKh, Prevars'ka N, Skryma R, Shuba IaM. The influence of extracellular pH on volume-activated chloride current in the prostate cancer epithelial cells. Fiziol Zh. 2002;48(4):19-27. Ukrainian.



Involvement of thermo- and mechanosensitive ion channels in the functioning of smooth muscles of the male urogenital system

The opening and closing (i.e. gating) of the most known types ion channels is governed by such classical stimuli as membrane potential and/or endogenous chemical ligands called neurotransmitters. However, since the end of 1990s many new types of ion channels which are gated by non-classical stimuli, including temperature changes and mechanical stress, have been discovered. In the framework of this research direction, we asked how the channels activated by heat (TRPV family), cold (TRPA1, TRPM8) or mechanical stress (PIEZO family) are involved in the regulation of contractile activity of smooth muscles (SM) of the male urogenital system – the detrusor, the prostate, the vas deferens and tunica dartos, which are distinguished by a high degree of sensitivity to the thermal and mechanical (stretching) insults. These studies are conducted within the institutional budgetary funding from the National Academy of Sciences of Ukraine (NASU), as well as funding from NASU's specialized programs such as "Genome" and "Biotechnology". In these studies we not only elucidate normal expression and function of the thermo- and mechano-gated channels in these SM tissues at the cellular and subcellular levels, but also try to get an idea on their possible changes as a result of complications caused by diabetes (see relevant publications).

References ►

  1. Philyppov IB, Paduraru ON, Gulak KL, Skryma R, Prevarskaya N, Shuba YM. TRPA1-dependent regulation of bladder detrusor smooth muscle contractility in normal and type I diabetic rats. J Smooth Muscle Res. 2016;52:1-17.
  2. Vanden Abeele F, Kondratskyi A, Dubois C, Shapovalov G, Gkika D, Busserolles J, Shuba Y, Skryma R, Prevarskaya N. Complex modulation of the cold receptor TRPM8 by volatile anaesthetics and its role in complications of general anaesthesia. J Cell Sci. 2013 Oct 1;126(Pt 19):4479-89.
  3. Philyppov IB, Paduraru ON, Andreev YA, Grishin EV, Shuba YM. Modulation of TRPV1-dependent contractility of normal and diabetic bladder smooth muscle by analgesic toxins from sea anemone Heteractis crispa. Life Sci. 2012 Nov 2;91(19-20):912-20.
  4. Skryma R, Prevarskaya N, Gkika D, Shuba Y. From urgency to frequency: facts and controversies of TRPs in the lower urinary tract. Nat Rev Urol. 2011 Oct 4;8(11):617-30. Review.
  5. Vladymyrova IA, Filippov IB, Kuliieva IeM, Iurkevych A, Skryma R, Prevarskaia N, Shuba IaM. Comparative effects of menthol and icilin on the induced contraction of the smooth muscles of the vas deferens of normal and castrated rats. Fiziol Zh. 2011;57(4):21-33. Ukrainian.
  6. Paduraru OM, Filippov IB, Boldyriev OI, Vladymyrova IA, Naĭd'onov VH, Shuba IaM. Urothelium-dependent modulation of urinary bladder smooth muscle contractions by menthol. Fiziol Zh. 2011;57(6):15-22. Ukrainian.
  7. Boldyrev OI, Sotkis HV, Kuliieva IeM, Vladymyrova IA, Filippov IB, Skryma R, Prevars'ka N, Shuba IaM. Expression of the cold receptor TRPM8 in the smooth muscles of the seminal ejaculatory ducts in rats. Fiziol Zh. 2009;55(5):17-27. Ukrainian.
  8. Filippov IB, Vladymyrova IA, Kuliieva IeM, Skryma R, Prevarskaia N, Shuba IaM. Modulation of the smooth muscle contractions of the rat vas deferens by TRPM8 channel agonist menthol. Fiziol Zh. 2009;55(6):30-40. Ukrainian.
  9. Kondrats'kyĭ AP, Kondrats'ka KO, Skryma R, Prevars'ka N, Shuba IaM. Gender differences in cold sensitivity: role of hormonal regulation of TRPM8 channel. Fiziol Zh. 2009;55(4):91-9. Ukrainian.
  10. Kondrats'kyĭ AP, Sotkis HV, Boldyriev OI, Kondrats'ka KO, Liubanova OP, Dyskina IuB, Hordiienko DV, Shuba IaM. Functional identification of the TRPM8 cold receptor in rat prostate epithelial cells. Fiziol Zh. 2007;53(5):3-13. Ukrainian.



The influence of diabetes on neuromuscular transmission and excitation-contraction coupling in smooth muscles of the male urogenital system

The impairment of the autonomic nervous system, peripheral diabetic neuropathy, is a frequent and serious complication of diabetes mellitus (DM). The weakening of the autonomic regulation of the lower urinary tract in patients with DM provokes the development of diabetic cystopathies, the main symptom of which is urinary incontinence. However, literature data on the impact of DM on neuromuscular synaptic transmission in visceral smooth muscle, and in particular the bladder detrusor smooth muscle (DSM), are quite controversial. The dominant view is that in diabetes, the expression of muscarinic m-cholinergic receptors (mAChR) and DSM sensitivity to their agonists is increased. Moreover, mAChR antagonists are considered to be effective in alleviating the symptoms of hyperactive urinary bladder and urge incontinence of different etiologies, although such therapy should be applied with caution due to significant side effects. Recently, potentially new determinants of bladder hyperactivity amongst ion channels from Transient Receptor Potential (TRP) family, TRPV1 and TRPA1, acting as receptors of burning and cooling temperatures, respectively, have been identified. The degree of severity and direction of urological complications in diabetes can also largely depend on inflammation and/or infection of the lower urinary tract that often accompany this disease. In this regard, we aimed to investigate how TRPV1 and TRPA1 are involved in changes of DSM contractility in diabetes and how DSM contractility in diabetic bladder is influenced by accompanying inflammation or infection. These studies are carried out within the institutional budgetary funding from the National Academy of Sciences of Ukraine (NASU), as well as funding from NASU's specialized "Genome" program (see relevant publications).

References ►

  1. Philyppov IB, Paduraru ON, Gulak KL, Skryma R, Prevarskaya N, Shuba YM. TRPA1-dependent regulation of bladder detrusor smooth muscle contractility in normal and type I diabetic rats. J Smooth Muscle Res. 2016;52:1-17.
  2. Vladimirova IA, Philyppov IB, Kulieva EM, Paduraru ON, Shuba YY, Shuba YM. Impact of diabetic complications on neuromuscular transmission in the smooth muscle of the bladder of rats with experimental diabetes. Fiziol Zh. 2015;61(4):56-62. Ukrainian.
  3. Vladimirova IA, Lankin YN, Philyppov IB, Sushiy LF, Shuba YM. Frequency dependence of excitation-contraction of multicellular smooth muscle preparations: the relevance to bipolar electrosurgery. J Surg Res. 2014 Jan;186(1):119-25.
  4. Vladimirova IA, Filippov IB, Paduraru ON, Shuba ÉIa, Kuliieva ÉM, Shuba IaM. Changes of neuromuscular transmission in smooth muscles of rats bladder with experimental diabetes. Fiziol Zh. 2014;60(2):31-7. Ukrainian.
  5. Philyppov IB, Paduraru ON, Andreev YA, Grishin EV, Shuba YM. Modulation of TRPV1-dependent contractility of normal and diabetic bladder smooth muscle by analgesic toxins from sea anemone Heteractis crispa. Life Sci. 2012 Nov 2;91(19-20):912-20.
  6. Kryshtal' DA, Paduraru OM, Boldyriev OI, Kit OIu, Rekalov VV, Shuba IaM. Changes in calcium-dependent potassium channels of isolated smooth muscle cells of the bladder in rats with experimental diabetes. Fiziol Zh. 2011;57(3):25-32. Ukrainian.



Targets of pharmacological action of new openers of ATP-dependent potassium channels (KATP) and their selectivity towards tissue-specific KATP subtypes of these channels

ATP-dependent potassium channels (KATP) play pivotal role in the coupling of physiological activity of cells with their intracellular energetics: when cell energy metabolism is compromised (i.e., the intracellular ATP to ADP ration is low), these channels become activated, leading to hyperpolarization of the membrane potential and suppression of cellular activity (i.e., the cell is turned into resting mode of operation). Thus, pharmacological agents, the so-called KATP-openers, are potentially very important protective therapeutic agents. Depending on their subunit composition, KATP-s are subdivided into three tissue-specific subtypes, cardiac, vascular and pancreatic. In the framework of this research direction, we investigate the peculiarities of the pharmacological action of the newly developed Ukrainian KATP-opener, flocalin. Flocalin is fluorine-containing derivative of a well known KATP-opener, pinacidil, which is positioned as cardioprotective agent against ischemic heart injury. Our data show that flocalin is indeed an effective opener of cardiac-type KATP-s, and that this effect underlies its cardioprotective properties. However, flocalin is also able to partially block sodium and calcium channels, which should be considered when establishing its therapeutic doses. The data obtained in native cardiomyocytes and bladder smooth muscle cells have already been published (see relevant publications). As a next step we aim to test the specificity of the flocalin action on different subtypes of recombinant, heterologously expressed KATP-channels.

References ►

  1. Philyppov IB, Golub AА, Boldyriev OI, Shtefan NL, Totska K, Voitychuk OI, Shuba YM. Myorelaxant action of fluorine-containing pinacidil analog, flocalin, in bladder smooth muscle is mediated by inhibition of L-type calcium channels rather than activation of KATP channels. Naunyn Schmiedebergs Arch Pharmacol. 2016 Jun;389(6):585-92.
  2. Al Kury LT, Voitychuk OI, Yang KH, Thayyullathil FT, Doroshenko P, Ramez AM, Shuba YM, Galadari S, Howarth FC, Oz M. Effects of the endogenous cannabinoid anandamide on voltage-dependent sodium and calcium channels in rat ventricular myocytes. Br J Pharmacol. 2014 Jul;171(14):3485-98.
  3. Voitychuk OI, Strutynskyi RB, Moibenko OO, Shuba YM. Effects of fluorine-containing opener of ATP-sensitive potassium channels, pinacidil-derivative flocalin, on cardiac voltage-gated sodium and calcium channels. Naunyn Schmiedebergs Arch Pharmacol. 2012 Nov;385(11):1095-102.
  4. Voitychuk OI, Asmolkova VS, Gula NM, Sotkis GV, Galadari S, Howarth FC, Oz M, Shuba YM. Modulation of excitability, membrane currents and survival of cardiac myocytes by N-acylethanolamines. Biochim Biophys Acta. 2012 Sep;1821(9):1167-76.
  5. Voitychuk OI, Strutynskyi RB, Yagupolskii LM, Tinker A, Moibenko OO, Shuba YM. Sarcolemmal cardiac K(ATP) channels as a target for the cardioprotective effects of the fluorine-containing pinacidil analogue, flocalin. Br J Pharmacol. 2011 Feb;162(3):701-11.
  6. Voĭtychuk OI, Asmolkova VS, Hula NM, Oz M, Shuba IaM. Effects of N-stearoyl- and N-oleoylethanolamine on cardiac voltage-dependent sodium channels. Fiziol Zh. 2010;56(5):13-22. Ukrainian.
  7. Voĭtychuk OI, Asmolkova VS, Hula NM, Sotkis HV, Oz M, Shuba IaM. Regulation of the excitability of neonatal cardiomyocytes by N-stearoyl- and N-oleoyl-ethanolamines. Fiziol Zh. 2009;55(3):55-66. Ukrainian.



Expression, biophysical properties, pharmacology and epigenetic regulation of low-threshold calcium channels in norm and in experimental abscess epilepsy

Low voltage-activated (LVA) calcium channels, also referred to as T-type calcium channels, are characterized by a number of biophysical properties that significantly differentiate them from other types of voltage-gated calcium channels (VGCCs). Three subtypes of LVA channels, namely Cav3.1, Cav3.2 and Cav3.3, have been identified on molecular level, each of which is encoded by a separate gene with specific expression in various tissues and cell types. Accordingly, the functional significance of each LVA-channel subtype may be quite unique and not always obvious, which makes the study of these channels very important. In our interest to the LVA calcium channels, we focused on three main directions:

  1. The study of how LVA channels discriminate various polyvalent cations in terms of either passing them through or being blocked by them;
  2. Determining potential factors that may regulate expression of LVA-channel subtypes;
  3. Establishing the role of LVA channels in epileptogenesis.

Our data obtained on Cav3.1, Cav3.2, Cav3.3 channels heterologously expressed in Xenopus oocytes revealed that these channels nearly similar pass Ca2+, Sr2+ and Ba2+, and that under physiological conditions they are also noticeably permeable to Na+. Blockade of these channels by some of the transition polyvalent cations, particularly Ni2+, require interaction with at least three channel's binding sites. Studies of different LVA-channel subtypes in the thalamus nuclei of normal rats in ontogenesis and rats with absence epilepsy phenotype (WAG/Rij strain) showed that the latter is accompanied by an increase in the expression of predominantly Cav3.1 channel and that the regulation of expression of various subtypes, both in ontogenesis and in pathologies, may be regulated by microRNA, miR-1 (see relevant publications).

References ►

  1. Sharop BR, Boldyriev OI, Batiuk MY, Shtefan NL, Shuba YM. Compensatory reduction of Cav3.1 expression in thalamocortical neurons of juvenile rats of WAG/Rij model of absence epilepsy. Epilepsy Res. 2016 Jan;119:10-2.
  2. Shuba YM. Models of calcium permeation through T-type channels. Pflugers Arch. 2014 Apr;466(4):635-44. Review.
  3. Nosal OV, Lyubanova OP, Naidenov VG, Shuba YM. Complex modulation of Ca(v)3.1 T-type calcium channel by nickel. Cell Mol Life Sci. 2013 May;70(9):1653-61.
  4. Shuba YM, Perez-Reyes E, Lory P, Noebels J. T-type calcium channels: from discovery to channelopathies, 25 years of research. Channels (Austin). 2008 Jul-Aug;2(4):299-302. Meeting report.
  5. Shcheglovitov A, Kostyuk P, Shuba Y. Selectivity signatures of three isoforms of recombinant T-type Ca2+ channels. Biochim Biophys Acta. 2007 Jun;1768(6):1406-19.
  6. Shcheglovitov A, Zhelay T, Vitko Y, Osipenko V, Perez-Reyes E, Kostyuk P, Shuba Y. Contrasting the effects of nifedipine on subtypes of endogenous and recombinant T-type Ca2+ channels. Biochem Pharmacol. 2005 Mar 1;69(5):841-54.
  7. Dosenko VE, Lyubanova OP, Shcheglovitov AK, Boldyryev AI, Shuba YaM. [Expression of RNA of subunits of low-threshold calcium channels in the laterodorsal nucleus of the rat thalamus: an ontogenetic aspect]. Neurophysiology (Kiev) 2005 May 37(3):201-5. Russian.
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