Biología molecular de los factores de virulencia de Entamoeba histolytica: detectamos genes y proteínas que participan en la adhesión, fagocitosis y destrucción celular producida por esta amiba.  Una proteína de 124 kDa localizada en la superficie de los trofozoítos, es codificada por dos genes adyacentes denominados Ehadh y Ehcp112. Este complejo EhCPADH contribuye con la adhesión de los parásitos a sus células blanco, durante la eritrofagocitosis y también presenta actividad de proteasa, lo que le permite dañar las uniones intercelulares de las células epiteliales. Los genes que codifican para este complejo han sido clonados en vectores especiales y ofrecen una alternativa como candidatos potenciales para el desarrollo de vacunas contra este parásito.

Recientemente, identificamos nuevos miembros de la maquinaria ESCRT, proteínas Rab y lípidos como LBPA y colesterol, así como proteínas de tráfico de colesterol, que participan en la vía endocítica de este parásito y actualmente estamos caracterizando sus funciones durante los diferentes eventos patogénicos.

Biología molecular y genética de la multirresistencia a fármacos en E. histolytica: estudiamos los genes y las proteínas involucradas en la multirresistencia a fármacos, desarrollando trofozoítos mutantes y resistentes a diferentes drogas. Hemos detectado, clonado y caracterizado genes como Ehpgp5 y Ehpgp1, los cuales son inducibles por emetina y constitutivo, respectivamente. También estamos analizado los promotores de esos genes y cómo participan diversos factores nucleares en su expresión.

Organización del genoma amibiano: caracterizamos el careotipo molecular of E. histolytica. En el citoplasma esta amiba también presenta DNA contenido en organelos que nombramos como EhkO. Recientemente, estamos estudiando el intercambio de material genético entre los trofozoítos provenientes del mismo cultivo o entre diferentes cepas. Además, analizamos los factores involucrados en este intercambio. Los hallazgos podrían explicar la diversidad que presentan las diferentes cepas de E. histolytica.

• Avalos-Padilla Y, Georgiev VN, Ewins E, Robinson T, Orozco E, Lipowsky R, Dimova R. Stepwise remodeling and subcompartment formation in individual vesicles by three ESCRT-III proteins. iScience. 2022 Dec 8;26(1):105765. doi: 10.1016/j.isci.2022.105765. PMID: 36590172; PMCID: PMC9800321.
• Bañuelos C, Betanzos A, Javier-Reyna R, Galindo A, Orozco E. Molecular interplays of the Entamoeba histolytica endosomal sorting complexes required for transport during phagocytosis. Front Cell Infect Microbiol. 2022 Oct 27;12:855797. doi: 10.3389/fcimb.2022.855797. PMID: 36389174; PMCID: PMC9647190.
• Galindo A, Javier-Reyna R, García-Rivera G, Bañuelos C, Montaño S, Ortega-Lopez J, Chávez-Munguía B, Salazar-Villatoro L, Orozco E. EhVps23: A component of ESCRT-I that participates in vesicular trafficking and phagocytosis of Entamoeba histolytica. Front Cell Infect Microbiol. 2021 Oct 29;11:770759. doi: 10.3389/fcimb.2021.770759. PMID: 34778112; PMCID: PMC8588831.
• Orozco, E., Betanzos, A., Bañuelos, C., Javier-Reyna, R., García-Rivera, G. (2020). Vesicular trafficking in Entamoeba histolytica is essential for its virulence. In: Guillen, N. (eds) Eukaryome impact on human intestine homeostasis and mucosal immunology. Springer, Cham. https://doi.org/10.1007/978-3-030-44826-4_20
• Javier-Reyna R, Montaño S, García-Rivera G, Rodríguez MA, González-Robles A, Orozco E. EhRabB mobilises the EhCPADH complex through the actin cytoskeleton during phagocytosis of Entamoeba histolytica. Cell Microbiol. 2019 Oct;21(10):e13071. doi: 10.1111/cmi.13071. Epub 2019 Jul 7. PMID: 31219662.
• Betanzos A, Bañuelos C, Orozco E. Host invasion by pathogenic amoebae: epithelial disruption by parasite proteins. Genes (Basel). 2019 Aug 14;10(8):618. doi: 10.3390/genes10080618. PMID: 31416298; PMCID: PMC6723116.
• Narayanasamy RK, Castañón-Sanchez CA, Luna-Arias JP, García-Rivera G, Avendaño-Borromeo B, Labra-Barrios ML, Valdés J, Herrera-Aguirre ME, Orozco E. The Entamoeba histolytica TBP and TRF1 transcription factors are GAAC-box binding proteins, which display differential gene expression under different stress stimuli and during the interaction with mammalian cells. Parasit Vectors. 2018 Mar 7;11(1):153. doi: 10.1186/s13071-018-2698-7. PMID: 29514716; PMCID: PMC5842622.
• Montaño S, Orozco E, Correa-Basurto J, Bello M, Chávez-Munguía B, Betanzos A. Heterodimerization of the Entamoeba histolytica EhCPADH virulence complex through molecular dynamics and protein-protein docking. J Biomol Struct Dyn. 2017 Feb;35(3):486-503. doi: 10.1080/07391102.2016.1151831. Epub 2016 Apr 18. PMID: 26861050.
• Bolaños J, Betanzos A, Javier-Reyna R, García-Rivera G, Huerta M, Pais-Morales J, González-Robles A, Rodríguez MA, Schnoor M, Orozco E. EhNPC1 and EhNPC2 Proteins participate in trafficking of exogenous cholesterol in Entamoeba histolytica trophozoites: Relevance for phagocytosis. PLoS Pathog. 2016 Dec 21;12(12):e1006089. doi: 10.1371/journal.ppat.1006089. PMID: 28002502; PMCID: PMC5176366.
• Pais-Morales J, Betanzos A, García-Rivera G, Chávez-Munguía B, Shibayama M, Orozco E. Resveratrol induces apoptosis-like death and prevents in vitro and in vivo virulence of Entamoeba histolytica. PLoS One. 2016 Jan 5;11(1):e0146287. doi: 10.1371/journal.pone.0146287. PMID: 26731663; PMCID: PMC4701480.
• Debnath A, Parsonage D, Andrade RM, He C, Cobo ER, Hirata K, Chen S, García-Rivera G, Orozco E, Martínez MB, Gunatilleke SS, Barrios AM, Arkin MR, Poole LB, McKerrow JH, Reed SL. A high-throughput drug screen for Entamoeba histolytica identifies a new lead and target. Nat Med. 2012 Jun;18(6):956-60. doi: 10.1038/nm.2758. PMID: 22610278; PMCID: PMC3411919.
• López-Reyes I, García-Rivera G, Bañuelos C, Herranz S, Vincent O, López-Camarillo C, Marchat LA, Orozco E. Detection of the endosomal sorting complex required for transport in Entamoeba histolytica and characterization of the EhVps4 protein. J Biomed Biotechnol. 2010;2010:890674. doi: 10.1155/2010/890674. Epub 2010 May 17. PMID: 20508821; PMCID: PMC2875786.

• Avalos-Padilla Y, Georgiev VN, Ewins E, Robinson T, Orozco E, Lipowsky R, Dimova R. Stepwise remodeling and subcompartment formation in individual vesicles by three ESCRT-III proteins. iScience. 2022 Dec 8;26(1):105765. doi: 10.1016/j.isci.2022.105765. PMID: 36590172; PMCID: PMC9800321.
• Bañuelos C, Betanzos A, Javier-Reyna R, Galindo A, Orozco E. Molecular interplays of the Entamoeba histolytica endosomal sorting complexes required for transport during phagocytosis. Front Cell Infect Microbiol. 2022 Oct 27;12:855797. doi: 10.3389/fcimb.2022.855797. PMID: 36389174; PMCID: PMC9647190.
• Galindo A, Javier-Reyna R, García-Rivera G, Bañuelos C, Chávez-Munguía B, Salazar-Villatoro L, Orozco E. EhVps23, an ESCRT-I member, is a key factor in secretion, motility, phagocytosis and tissue invasion by Entamoeba histolytica. Front Cell Infect Microbiol. 2022 Mar 14;12:835654. doi: 10.3389/fcimb.2022.835654. PMID: 35360117; PMCID: PMC8964110.
• Galindo A, Javier-Reyna R, García-Rivera G, Bañuelos C, Montaño S, Ortega-Lopez J, Chávez-Munguía B, Salazar-Villatoro L, Orozco E. EhVps23: A component of ESCRT-I that participates in vesicular trafficking and phagocytosis of Entamoeba histolytica. Front Cell Infect Microbiol. 2021 Oct 29;11:770759. doi: 10.3389/fcimb.2021.770759. PMID: 34778112; PMCID: PMC8588831.
• Díaz-Hernández M, Javier-Reyna R, Sotto-Ortega I, García-Rivera G, Montaño S, Betanzos A, Zanatta D, Orozco E. Protein sumoylation is crucial for phagocytosis in Entamoeba histolytica Trophozoites. Int J Mol Sci. 2021 May 27;22(11):5709. doi: 10.3390/ijms22115709. PMID: 34071922; PMCID: PMC8198320.
• Orozco, E., Betanzos, A., Bañuelos, C., Javier-Reyna, R., García-Rivera, G. (2020). Vesicular trafficking in Entamoeba histolytica is essential for its virulence. In: Guillen, N. (eds) Eukaryome impact on human intestine homeostasis and mucosal immunology. Springer, Cham. https://doi.org/10.1007/978-3-030-44826-4_20
• Huerta M, Reyes L, García-Rivera G, Bañuelos C, Betanzos A, Díaz-Hernández M, Galindo A, Bolaños J, Cárdenas H, Azuara-Liceaga E, Chávez-Munguía B, Orozco E. A noncanonical GATA transcription factor of Entamoeba histolytica modulates genes involved in phagocytosis. Mol Microbiol. 2020 Dec;114(6):1019-1037. doi: 10.1111/mmi.14592. Epub 2020 Sep 10. PMID: 32808689.
• Castellanos-Castro S, Bolaños J, Orozco E. Lipids in Entamoeba histolytica: host-dependence and virulence factors. Front Cell Infect Microbiol. 2020 Mar 10;10:75. doi: 10.3389/fcimb.2020.00075. PMID: 32211340; PMCID: PMC7075943.
• Javier-Reyna R, Montaño S, García-Rivera G, Rodríguez MA, González-Robles A, Orozco E. EhRabB mobilises the EhCPADH complex through the actin cytoskeleton during phagocytosis of Entamoeba histolytica. Cell Microbiol. 2019 Oct;21(10):e13071. doi: 10.1111/cmi.13071. Epub 2019 Jul 7. PMID: 31219662.
• Betanzos A, Bañuelos C, Orozco E. Host invasion by pathogenic amoebae: epithelial disruption by parasite proteins. Genes (Basel). 2019 Aug 14;10(8):618. doi: 10.3390/genes10080618. PMID: 31416298; PMCID: PMC6723116.
• Avendaño-Borromeo B, Narayanasamy RK, García-Rivera G, Labra-Barrios ML, Lagunes-Guillén AE, Munguía-Chávez B, Castañón-Sánchez CA, Orozco E, Luna-Arias JP. Identification of the gene encoding the TATA box-binding protein-associated factor 1 (TAF1) and its putative role in the heat shock response in the protozoan parasite Entamoeba histolytica. Parasitol Res. 2019 Feb;118(2):517-538. doi: 10.1007/s00436-018-6170-6. Epub 2018 Dec 15. PMID: 30552577.
• Rendón-Gandarilla FJ, Álvarez-Hernández V, Castañeda-Ortiz EJ, Cárdenas-Hernández H, Cárdenas-Guerra RE, Valdés J, Betanzos A, Chávez-Munguía B, Lagunes-Guillen A, Orozco E, López-Canovas L, Azuara-Liceaga E. Telomeric repeat-binding factor homologs in Entamoeba histolytica: new clues for telomeric research. Front Cell Infect Microbiol. 2018 Oct 2;8:341. doi: 10.3389/fcimb.2018.00341. PMID: 30333961; PMCID: PMC6175992.
• Betanzos A, Zanatta D, Bañuelos C, Hernández-Nava E, Cuellar P, Orozco E. Epithelial cells expressing EhADH, an Entamoeba histolytica adhesin, exhibit increased tight junction proteins. Front Cell Infect Microbiol. 2018 Sep 28;8:340. doi: 10.3389/fcimb.2018.00340. PMID: 30324093; PMCID: PMC6172307.
• Azuara-Liceaga E, Betanzos A, Cardona-Felix CS, Castañeda-Ortiz EJ, Cárdenas H, Cárdenas-Guerra RE, Pastor-Palacios G, García-Rivera G, Hernández-Álvarez D, Trasviña-Arenas CH, Diaz-Quezada C, Orozco E, Brieba LG. The Sole DNA ligase in Entamoeba histolytica is a high-fidelity DNA ligase involved in DNA damage repair. Front Cell Infect Microbiol. 2018 Jul 12;8:214. doi: 10.3389/fcimb.2018.00214. Erratum in: Front Cell Infect Microbiol. 2022 Dec 16;12:1023564. PMID: 30050869; PMCID: PMC6052137.
• Avalos-Padilla Y, Knorr RL, Javier-Reyna R, García-Rivera G, Lipowsky R, Dimova R, Orozco E. The conserved ESCRT-III machinery participates in the phagocytosis of Entamoeba histolytica. Front Cell Infect Microbiol. 2018 Mar 1;8:53. doi: 10.3389/fcimb.2018.00053. PMID: 29546036; PMCID: PMC5838018.
• Narayanasamy RK, Castañón-Sanchez CA, Luna-Arias JP, García-Rivera G, Avendaño-Borromeo B, Labra-Barrios ML, Valdés J, Herrera-Aguirre ME, Orozco E. The Entamoeba histolytica TBP and TRF1 transcription factors are GAAC-box binding proteins, which display differential gene expression under different stress stimuli and during the interaction with mammalian cells. Parasit Vectors. 2018 Mar 7;11(1):153. doi: 10.1186/s13071-018-2698-7. PMID: 29514716; PMCID: PMC5842622.
• Cuellar P, Hernández-Nava E, García-Rivera G, Chávez-Munguía B, Schnoor M, Betanzos A, Orozco E. Entamoeba histolytica EhCP112 dislocates and degrades claudin-1 and claudin-2 at tight junctions of the intestinal epithelium. Front Cell Infect Microbiol. 2017 Aug 16;7:372. doi: 10.3389/fcimb.2017.00372. PMID: 28861400; PMCID: PMC5561765.
• Hernández-Nava E, Cuellar P, Nava P, Chávez-Munguía B, Schnoor M, Orozco E, Betanzos A. Adherens junctions and desmosomes are damaged by Entamoeba histolytica: Participation of EhCPADH complex and EhCP112 protease. Cell Microbiol. 2017 Nov;19(11). doi: 10.1111/cmi.12761. Epub 2017 Jul 18. PMID: 28656597.
• Montaño S, Orozco E, Correa-Basurto J, Bello M, Chávez-Munguía B, Betanzos A. Heterodimerization of the Entamoeba histolytica EhCPADH virulence complex through molecular dynamics and protein-protein docking. J Biomol Struct Dyn. 2017 Feb;35(3):486-503. doi: 10.1080/07391102.2016.1151831. Epub 2016 Apr 18. PMID: 26861050.
• Bolaños J, Betanzos A, Javier-Reyna R, García-Rivera G, Huerta M, Pais-Morales J, González-Robles A, Rodríguez MA, Schnoor M, Orozco E. EhNPC1 and EhNPC2 Proteins participate in trafficking of exogenous cholesterol in Entamoeba histolytica trophozoites: Relevance for phagocytosis. PLoS Pathog. 2016 Dec 21;12(12):e1006089. doi: 10.1371/journal.ppat.1006089. PMID: 28002502; PMCID: PMC5176366.
• Pais-Morales J, Betanzos A, García-Rivera G, Chávez-Munguía B, Shibayama M, Orozco E. Resveratrol induces apoptosis-like death and prevents in vitro and in vivo virulence of Entamoeba histolytica. PLoS One. 2016 Jan 5;11(1):e0146287. doi: 10.1371/journal.pone.0146287. PMID: 26731663; PMCID: PMC4701480.
• Castellanos-Castro S, Cerda-García-Rojas CM, Javier-Reyna R, Pais-Morales J, Chávez-Munguía B, Orozco E. Identification of the phospholipid lysobisphosphatidic acid in the protozoan Entamoeba histolytica: An active molecule in endocytosis. Biochem Biophys Rep. 2015 Dec 23;5:224-236. doi: 10.1016/j.bbrep.2015.12.010. PMID: 28955828; PMCID: PMC5600446.
• Avalos-Padilla Y, Betanzos A, Javier-Reyna R, García-Rivera G, Chávez-Munguía B, Lagunes-Guillén A, Ortega J, Orozco E. EhVps32 is a vacuole-associated protein involved in pinocytosis and phagocytosis of Entamoeaba histolytica. PLoS Pathog. 2015 Jul 31;11(7):e1005079. doi: 10.1371/journal.ppat.1005079. PMID: 26230715; PMCID: PMC4521941.
• Borbolla-Vázquez J, Orozco E, Betanzos A, Rodríguez MA. Entamoeba histolytica: protein arginine transferase 1a methylates arginine residues and potentially modify the H4 histone. Parasit Vectors. 2015 Apr 10;8:219. doi: 10.1186/s13071-015-0820-7. PMID: 25889855; PMCID: PMC4393863.
• Betanzos A, Schnoor M, Javier-Reyna R, García-Rivera G, Bañuelos C, Pais-Morales J, Orozco E. Analysis of the epithelial damage produced by Entamoeba histolytica infection. J Vis Exp. 2014 Jun 12;(88):51668. doi: 10.3791/51668. PMID: 24962382; PMCID: PMC4189439.
• Betanzos A, Javier-Reyna R, García-Rivera G, Bañuelos C, González-Mariscal L, Schnoor M, Orozco E. The EhCPADH112 complex of Entamoeba histolytica interacts with tight junction proteins occludin and claudin-1 to produce epithelial damage. PLoS One. 2013 Jun 7;8(6):e65100. doi: 10.1371/journal.pone.0065100. PMID: 23762290; PMCID: PMC3676397.
• Debnath A, Parsonage D, Andrade RM, He C, Cobo ER, Hirata K, Chen S, García-Rivera G, Orozco E, Martínez MB, Gunatilleke SS, Barrios AM, Arkin MR, Poole LB, McKerrow JH, Reed SL. A high-throughput drug screen for Entamoeba histolytica identifies a new lead and target. Nat Med. 2012 Jun;18(6):956-60. doi: 10.1038/nm.2758. PMID: 22610278; PMCID: PMC3411919.
• Bañuelos C, García-Rivera G, López-Reyes I, Mendoza L, González-Robles A, Herranz S, Vincent O, Orozco E. EhADH112 is a Bro1 domain-containing protein involved in the Entamoeba histolytica multivesicular bodies pathway. J Biomed Biotechnol. 2012;2012:657942. doi: 10.1155/2012/657942. Epub 2012 Feb 28. PMID: 22500103; PMCID: PMC3303925.
• López-Reyes I, García-Rivera G, Bañuelos C, Herranz S, Vincent O, López-Camarillo C, Marchat LA, Orozco E. Detection of the endosomal sorting complex required for transport in Entamoeba histolytica and characterization of the EhVps4 protein. J Biomed Biotechnol. 2010;2010:890674. doi: 10.1155/2010/890674. Epub 2010 May 17. PMID: 20508821; PMCID: PMC2875786.