Summary of the studies that describe the effect of
Abstract
Chagas disease affects low-income nations with health consequences that impact the economy of those countries. Interestingly, inhibitors of channels formed by proteins of the gap junction family, such as suramin and boldine, exhibit trypanocidal activity. Gap junction proteins are integral membrane proteins present in both vertebrates and invertebrates that participate in cellular communication. These proteins form gap junction channels, which connect the cytoplasm of neighboring cells or non-junctional channels that connect the intra- and extracellular milieu. Interestingly, Trypanosoma cruzi modulates the expression of proteins of the gap junction family or modify the activity of the channels formed by these proteins in host cells. Moreover, Lucifer yellow microinjected into fibroblast was incorporated into associated trypanosomes of Trypanosoma musculi, suggesting the possibility of direct communication via gap junction channels between them. In this chapter, we summarized the current knowledge about the possible roles of gap junction family proteins in Chagas disease.
Keywords
- connexin
- pannexin
- innexin
- hemichannels
- infection
1. Introduction
Chagas disease affects low-income nations with health consequences that impact the economy of these countries [1]. Research aimed at understanding their biology and identification of potential targets for drug development is the highest priority [1]. Interestingly, inhibitors of channels formed by proteins of the gap junction family such as suramin and boldine have trypanocidal activity and some of them are currently used for treatment of parasitic diseases such Human African Trypanosomiasis [2, 3, 4, 5]. Also, studies have shown that infections caused by
2. Gap junction proteins
Gap junction proteins are present in both vertebrates and invertebrates from mesozoa to mammals [13]. The protein families include connexins (Cxs), innexins (Inxs) and pannexins (Panxs) [14, 15]. They are integral membrane proteins that participate in cellular communication playing a relevant role in several physiological processes [15]. In vertebrates, Cxs and Panxs are present, while in invertebrates, only Inxs are present [15]. In humans, 21 and 3 genes encode Cx and Panx proteins, respectively [14]. Most Cx genes contain two exons and an intron of variable length [14]. The Panx1 and Panx2 genes contain 5 exons, while the Panx3 gene contains 4 exons [16]. Moreover, Inx genes have been found in the phylum
2.1 Gap junction channels
Cx and Inx proteins form gap junction channels, which connect the cytoplasm of neighboring cells [14, 15]. Moreover, Cx, Panx and Inx proteins form non-junctional channels that connect the intra- and extracellular milieu [14]. Gap junction plaques are formed by a variable number of homo- and/or heterotypic gap junction channels with distinct biophysical characteristics [29]. Structurally, they are formed by oligomers of Cx, Inx, or Panx proteins, which co-oligomerize into the same (homomeric) or mixed (heteromeric) channels [30]. Gap junction channels are essential in several physiologic functions such as electrical conduction between cardiomyocytes [31], development and regeneration of skeletal muscle [32], endocrine gland secretion [33], and ovarian folliculogenesis [34]. Also, they are implicated in pathophysiological conditions such as hereditary deafness [35], cataract [36], and tumorigenesis [37].
Gap junction proteins can also form non-junctional channels, which play important roles as autocrine/paracrine cellular communication pathways [14]. They are permeable to ions and several metabolic and signaling molecules such as glucose, glutamate, glutathione, adenosine, NAD+ (superindice) and ATP among others [14]. It has been proposed that Panxs do not form gap junctions, however, they form plasma membrane channels (named pannexons) with some properties similar to those of the non-junctional channels formed by Inxs or Cxs also called hemichannels [15].
The pannexons are permeable to ATP when are activated with certain stimuli such a low oxygen, mechanical stress, and elevated extracellular potassium ion concentration [38]. Otherwise, the Panx1 channel is selective for chloride ions and exhibits no ATP permeability when stimulated simply by depolarization to positive potentials or the C-terminal is cleaved by proteolysis [38]. Since we found increased Panx1 channel activity and increase in ATP release in cells infected with
Moreover, Cx-based hemichannels are regulated by intracellular acidification [43], intracellular Ca2+ [44], intracellular Na+ [45], pro-inflammatory cytokines [46], positive membrane potentials [47], phosphorylation [48], and S-nitrosylation [49]. They are inhibited by lanthanum chloride, carbenoxolone (>50 μM), and Cx mimetic peptides GAP16, GAP27, and GAP19 [15].
3. Gap junction and Chagas
3.1 Connexins
In 1992, Spray’s group showed for the first time that gap junctions were altered between rat neonatal cardiomyocytes infected with the Tulahuen strain of
3.2 Pannexins
In 2018, we found that H510 strain of
4. Gap junction channel blocking compounds as antiparasitic drugs
Interestingly, several drugs that block the activity of gap junction channels have been described as potent anti-parasitic agents. Among them are: probenecid, boldine and suramin, among others (Table 2) [2, 3, 4].
4.1 Probenecid
Human clinical trials have shown that probenecid, an inhibitor of a non-junctional channel formed by Panx1 has an antimalarial effect [2]. The authors described that 50 μM probenecid caused an increase in the sensitivity of highly resistant V1S strain of
4.2 Suramin
Suramin, a general antagonist of purinergic receptors (P2Y and P2X) [51] and blocker of Panx1 channels [52], exhibits anti-parasitic properties [3]. Culture of LLC-MK2 cells treated with suramin (500 μM) during the intracellular development of
4.3 Boldine
Boldine, an alkaloid obtained from Boldo tree, which blocks the activity of Cx43-formed hemichannels, and Panx1 hemichannels exhibit, an anti-parasitic activity [53, 54]. Boldine at concentrations above 500 μM reduces the epimastigotes growth of Tulahuen, LQ and DM28c strains of
5. Conclusions
Chagas diseases affect predominantly underprivileged areas of Latin America, but attention has been increasing lately due to the rise in people migration habits, intercontinental travels, and immune suppressed patients [1]. Unfortunately, current therapeutic options include only two compounds (nifurtimox and benznidazole) with considerable toxicity and side effects, so the new drug development is of the highest priority [1]. Hemichannels are involved in the regulation of plasma membrane permeability in ischemic insults or metabolic inhibition [55, 56]. Moreover, alterations of plasma membrane is a common phenomenon in parasite-induced infections such malaria and
Acknowledgments
This work was partially supported by a MINEDUC-UA project code ANT 1755 (to JLV), Fondo Nacional de Desarrollo Científico y Tecnológico (FONDECYT) 1150291 (to JCS) and ICM-Economía P09-022-F Centro Interdisciplinario de Neurociencias de Valparaíso (to JCS). JG and IB hold a CONICYT-Ph.D. fellowship, Chile.
Conflict of interest
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
References
- 1.
Rassi A Jr, Rassi A, Marin-Neto JA. Chagas disease. Lancet. 2010; 375 :1388-1402. DOI: 10.1016/S0140-6736(10)60061-X - 2.
Masseno V, Muriithi S, Nzila A. In vitro chemosensitization of Plasmodium falciparum to antimalarials by verapamil and probenecid. Antimicrobial Agents and Chemotherapy. 2009;53 :3131-3134. DOI: 10.1128/AAC.01689-08 - 3.
Bisaggio DF, Campanati L, Pinto RC, Souto-Padrón T. Effect of suramin on trypomastigote forms of Trypanosoma cruzi : Changes on cell motility and on the ultrastructure of the flagellum-cell body attachment region. Acta Tropica. 2006;98 :162-175. DOI: 10.1016/j.actatropica.2006.04.003 - 4.
Morello A, Lipchenca I, Cassels BK, Speisky H, Aldunate J, Repetto Y. Trypanocidal effect of boldine and related alkaloids upon several strains of Trypanosoma cruzi . Comparative Biochemistry and Physiology Part C: Pharmacology, Toxicology and Endocrinology. 1994;107 :367-371. DOI: 10.1016/1367-8280(94)90063-9 - 5.
Singh Grewal A, Pandita D, Bhardwaj S, Lather V. Recent updates on development of drug molecules for human African trypanosomiasis. Current Topics in Medicinal Chemistry. 2016; 16 (20):2245-2265 - 6.
de Carvalho AC, Tanowitz HB, Wittner M, Dermietzel R, Roy C, Hertzberg EL, et al. Gap junction distribution is altered between cardiac myocytes infected with Trypanosoma cruzi . Circulation Research. 1992;70 :733-742 - 7.
Campos de Carvalho AC, Roy C, Hertzberg EL, Tanowitz HB, Kessler JA, Weiss LM, et al. Gap junction disappearance in astrocytes and leptomeningeal cells as a consequence of protozoan infection. Brain Research. 1998; 790 :304-314. DOI: 10.1016/S0006-8993(97)01523-0 - 8.
Adesse D, Garzoni LR, Huang H, Tanowitz HB, de Nazareth MM, Spray DC. Trypanosoma cruzi induces changes in cardiac connexin43 expression. Microbes and Infection. 2008;10 :21-28. DOI: 10.1016/j.micinf.2007.09.017 - 9.
Waghabi MC, Coutinho-Silva R, Feige JJ, Higuchi Mde L, Becker D, Burnstock G, et al. Gap junction reduction in cardiomyocytes following transforming growth factor-beta treatment and Trypanosoma cruzi infection. Memórias do Instituto Oswaldo Cruz. 2009;104 :1083-1090. DOI: 10.1590/S0074-02762009000800004 - 10.
Burke S, Nagajyothi F, Thi MM, Hanani M, Scherer PE, Tanowitz HB, et al. Adipocytes in both brown and white adipose tissue of adult mice are functionally connected via gap junctions: Implications for Chagas disease. Microbes and Infection. 2014; 16 :893-901. DOI: 10.1016/j.micinf.2014.08.006 - 11.
Barría I, Güiza J, Cifuentes F, Zamorano P, Sáez JC, González J, et al. Trypanosoma cruzi infection induces pannexin-1 channel opening in cardiac myocytes. The American Journal of Tropical Medicine and Hygiene. 2018;98 :105-112. DOI: 10.4269/ajtmh.17-0293 - 12.
Gugssa A, Lee CM, Gebru S, Desta D, Murray S, Baccetti B, et al. Co-culture of Trypanosoma musculi with spleen-derived adherent fibroblasts: Possible transfer of small molecules via connexons. Journal of Submicroscopic Cytology and Pathology. 2005;37 :223-229 - 13.
Meşe G, Richard G, White TW. Gap junctions: Basic structure and function. The Journal of Investigative Dermatology. 2007; 127 :2516-2524. DOI: 10.1038/sj.jid.5700770 - 14.
Saez JC, Berthoud VM, Branes MC, Martinez AD, Beyer EC. Plasma membrane channels formed by connexins: Their regulation and functions. Physiological Reviews. 2003; 83 :1359-1400. DOI: 10.1152/physrev.00007.2003 - 15.
Vega JL, Subiabre M, Figueroa F, Schalper KA, Osorio L, González J, et al. Role of gap junctions and hemichannels in parasitic infections. BioMed Research International. 2013; 2013 :589130. DOI: 10.1155/2013/589130 - 16.
Baranova A, Ivanov D, Petrash N, Pestova A, Skoblov M, Kelmanson I, et al. The mammalian pannexin family is homologous to the invertebrate innexin gap junction proteins. Genomics. 2004; 83 :706-716. DOI: 10.1016/j.ygeno.2003.09.025 - 17.
Ganfornina MD, Sánchez D, Herrera M, Bastiani MJ. Developmental expression and molecular characterization of two gap junction channel proteins expressed during embryogenesis in the grasshopper Schistocerca americana . Developmental Genetics. 1999;24 :137-150. DOI: 10.1002/(SICI)1520-6408(1999)24:1/2<137::AID-DVG13>3.0.CO;2-7 - 18.
Stebbings LA, Todman MG, Phillips R, Greer CE, Tam J, Phelan P, et al. Gap junctions in Drosophila : Developmental expression of the entire innexin gene family. Mechanisms of Development. 2002;113 :197-205. DOI: 10.1016/S0925-4773(02)00025-4 - 19.
Hong SM, Kang SW, Goo TW, Kim NS, Lee JS, Kim KA, et al. Two gap junction channel (innexin) genes of the Bombyx mori and their expression. Journal of Insect Physiology. 2008;54 :180-191. DOI: 10.1016/j.jinsphys.2007.09.002 - 20.
Calkins TL, Woods-Acevedo MA, Hildebrandt O, Piermarini PM. The molecular and immunochemical expression of innexins in the yellow fever mosquito, Aedes aegypti : Insights into putative life stage- and tissue-specific functions of gap junctions. Comparative Biochemistry and Physiology Part B: Biochemistry & Molecular Biology. 2015;183 :11-21. DOI: 10.1016/j.cbpb.2014.11.013 - 21.
Altun Z, Chen B, Wang Z, Hall D. High resolution map of Caenorhabditis elegans gap junction proteins. Developmental Dynamics. 2009;238 :1936-1950. DOI: 10.1002/dvdy.22025 - 22.
White T, Wang H, Mui R, Litteral J, Brink P. Cloning and functional expression of invertebrate connexins from Halocynthia pyriformis . FEBS Letters. 2004;577 :42-48. DOI: 10.1016/j.febslet.2004.09.071 - 23.
Kandarian B, Sethi J, Wu A, Baker M, Yazdani N, Kym E, et al. The medicinal leech genome encodes 21 innexin genes: Different combinations are expressed by identified central neurons. Development Genes and Evolution. 2012; 222 :29-44. DOI: 10.1007/s00427-011-0387-z - 24.
Zurabian R, Landa A, Robert L, Willms K. Immunolocalization of Taenia solium gap junction innexins. Parasitology. 2008;135 :1125-1131. DOI: 10.1017/S0031182008004629 - 25.
Takaku Y, Hwang JS, Wolf A, Böttger A, Shimizu H, David CN, et al. Innexin gap junctions in nerve cells coordinate spontaneous contractile behavior in Hydra polyps. Scientific Reports. 2014;4 :3573. DOI: 10.1038/srep03573 - 26.
Kelmanson IV, Shagin DA, Usman N, Matz MV, Lukyanov SA, Panchin YV. Altering electrical connections in the nervous system of the pteropod mollusc Clione limacina by neuronal injections of gap junction mRNA. The European Journal of Neuroscience. 2002;16 :2475-2476. DOI: 10.1046/j.1460-9568.2002.02423.x - 27.
Simonsen K, Moerman D, Naus C. Gap junctions in C. elegans . Frontiers in Physiology. 2014;5 :40. DOI: 10.3389/fphys.2014.00040 - 28.
Phelan P. Innexins: Members of an evolutionarily conserved family of gap-junction proteins. Biochimica et Biophysica Acta - Biomembranes. 2005; 1711 :225-245. DOI: 10.1016/j.bbamem.2004.10.004 - 29.
Barbe MT, Monyer H, Bruzzone R. Cell-cell communication beyond connexins: The pannexin channels. Physiology (Bethesda). 2006; 21 :103-114. DOI: 10.1152/physiol.00048.2005 - 30.
Falk MM, Buehler LK, Kumar NM, Gilula NB. Cell-free synthesis and assembly of connexins into functional gap junction membrane channels. The EMBO Journal. 1997; 16 :2703-2716. DOI: 10.1093/emboj/16.10.2703 - 31.
Kanno S, Saffitz JE. The role of myocardial gap junctions in electrical conduction and arrhythmogenesis. Cardiovascular Pathology. 2001; 10 :169-177. DOI: 10.1016/S1054-8807(01)00078-3 - 32.
Araya R, Eckardt D, Maxeiner S, Krüger O, Theis M, Willecke K, et al. Expression of connexins during differentiation and regeneration of skeletal muscle: Functional relevance of connexin43. Journal of Cell Science. 2005; 118 :27-37. DOI: 10.1242/jcs.01553 - 33.
Murray SA, Davis K, Gay V. ACTH and adrenocortical gap junctions. Microscopy Research and Technique. 2003; 61 :240-246. DOI: 10.1002/jemt.10332 - 34.
Gershon E, Plaks V, Dekel N. Gap junctions in the ovary: Expression, localization and function. Molecular and Cellular Endocrinology. 2008; 282 :18-25. DOI: 10.1016/j.mce.2007.11.001 - 35.
Kelsell DP, Dunlop J, Stevens HP, Lench NJ, Liang JN, Parry G, et al. Connexin 26 mutations in hereditary non-syndromic sensorineural deafness. Nature. 1997; 387 :80-83. DOI: 10.1038/387080a0 - 36.
Xia CH, Chang B, Derosa AM, Cheng C, White TW, Gong X. Cataracts and microphthalmia caused by a Gja8 mutation in extracellular loop 2. PLoS One. 2012; 7 :e52894. DOI: 10.1371/journal.pone.0052894 - 37.
Naus CC, Laird DW. Implications and challenges of connexin connections to cancer. Nature Reviews Cancer. 2010; 10 :435-441. DOI: 10.1038/nrc2841 - 38.
Wang J, Dahl G. Pannexin1: A multifunction and multiconductance and/or permeability membrane channel. American Journal of Physiology-Cell Physiology. 2018; 315 :C290-C299. DOI: 10.1152/ajpcell.00302.2017 - 39.
Bao L, Locovei S, Dahl G. Pannexin membrane channels are mechanosensitive conduits for ATP. FEBS Letters. 2004; 572 :565-568. DOI: 10.1016/j.febslet.2004.07.009 - 40.
Dahl G, Muller KJ. Innexin and pannexin channels and their signaling. FEBS Letters. 2014; 588 :1396-1402. DOI: 10.1016/j.febslet.2014.03.007 - 41.
Bao L, Samuels S, Locovei S, Macagno ER, Muller KJ, Dahl G. Innexins form two types of channels. FEBS Letters. 2007; 581 :5703-5708. DOI: 10.1016/j.febslet.2007.11.030 - 42.
Locovei S, Bao L, Dahl G. Pannexin 1 in erythrocytes: Function without a gap. Proceedings of the National Academy of Sciences of the United States of America. 2006; 103 :7655-7659. DOI: 10.1073/pnas.0601037103 - 43.
Delmar M, Coombs W, Sorgen P, Duffy HS, Taffet SM. Structural bases for the chemical regulation of Connexin43 channels. Cardiovascular Research. 2004; 62 :268-275. DOI: 10.1016/j.cardiores.2003.12.030 - 44.
Lopez W, Gonzalez J, Liu Y, Harris AL, Contreras JE. Insights on the mechanisms of Ca(2+) regulation of connexin26 hemichannels revealed by human pathogenic mutations (D50N/Y). The Journal of General Physiology. 2013; 142 :23-35. DOI: 10.1085/jgp.201210893 - 45.
Li F, Sugishita K, Su Z, Ueda I, Barry WH. Activation of connexin-43 hemichannels can elevate [Ca(2+)]i and [Na(+)]i in rabbit ventricular myocytes during metabolic inhibition. Journal of Molecular and Cellular Cardiology. 2001; 33 :2145-2155. DOI: 10.1006/jmcc.2001.1477 - 46.
Retamal MA, Froger N, Palacios-Prado N, Ezan P, Sáez PJ, Sáez JC, et al. Cx43 hemichannels and gap junction channels in astrocytes are regulated oppositely by proinflammatory cytokines released from activated microglia. The Journal of Neuroscience. 2007; 27 :13781-13792. DOI: 10.1523/JNEUROSCI.2042-07.2007 - 47.
Contreras JE, Saez JC, Bukauskas FF, Bennett MV. Gating and regulation of connexin 43 (Cx43) hemichannels. Proceedings of the National Academy of Sciences of the United States of America. 2003; 100 :11388-11393. DOI: 10.1073/pnas.1434298100 - 48.
Pogoda K, Kameritsch P, Retamal M, Vega JL. Regulation of gap junction channels and hemichannels by phosphorylation and redox changes: A revision. BMC Cell Biology. 2016; 17 (Suppl 1):11. DOI: 10.1186/s12860-016-0099-3 - 49.
Retamal MA, Cortes CJ, Reuss L, Bennett MV, Saez JC. S-nitrosylation and permeation through connexin 43 hemichannels in astrocytes: Induction by oxidant stress and reversal by reducing agents. Proceedings of the National Academy of Sciences of the United States of America. 2006; 103 :4475-4480. DOI: 10.1073/pnas.0511118103 - 50.
Martins RM, Covarrubias C, Rojas RG, Silber AM, Yoshida N. Use of L-proline and ATP production by Trypanosoma cruzi metacyclic forms as requirements for host cell invasion. Infection and Immunity. 2009;77 :3023-3032. DOI: 10.1128/IAI.00138-09 - 51.
Voogd TE, Vansterkenburg EL, Wilting J, Janssen LH. Recent research on the biological activity of suramin. Pharmacological Reviews. 1993; 45 :177-203 - 52.
Qiu F, Dahl G. A permeant regulating its permeation pore: Inhibition of pannexin 1 channels by ATP. American Journal of Physiology-Cell Physiology. 2009; 296 :C250-C255. DOI: 10.1152/ajpcell.00433.2008 - 53.
Hernández-Salinas R, Vielma AZ, Arismendi MN, Boric MP, Sáez JC, Velarde V. Boldine prevents renal alterations in diabetic rats. Journal Diabetes Research. 2013; 2013 :593672. DOI: 10.1155/2013/593672 - 54.
Yi C, Ezan P, Fernández P, Schmitt J, Sáez JC, Giaume C, et al. Inhibition of glial hemichannels by boldine treatment reduces neuronal suffering in a murine model of Alzheimer’s disease. Glia. 2017; 65 :1607-1625. DOI: 10.1002/glia.23182 - 55.
Contreras JE, Sánchez HA, Eugenin EA, Speidel D, Theis M, Willecke K, et al. Metabolic inhibition induces opening of unapposed connexin 43 gap junction hemichannels and reduces gap junctional communication in cortical astrocytes in culture. Proceedings of the National Academy of Sciences of the United States of America. 2002; 99 :495-500. DOI: 10.1073/pnas.012589799 - 56.
Thompson RJ, Zhou N, MacVicar BA. Ischemia opens neuronal gap junction hemichannels. Science. 2006; 312 :924-927. DOI: 10.1126/science.1126241 - 57.
Ginsberg H. Alterations caused by the intraerythrocytic malaria parasite in the permeability of its host cell membrane. Comparative Biochemistry and Physiology Part A: Comparative Physiology. 1990; 95 :31-39. DOI: 10.1016/0300-9629(90)90006-E - 58.
Fernandes MC, Andrews NW. Host cell invasion by Trypanosoma cruzi : A unique strategy that promotes persistence. FEMS Microbiology Reviews. 2012;36 :734-747. DOI: 10.1111/j.1574-6976.2012.00333.x