Ελληνικό Ινστιτούτο Παστέρ

The main research activity of the group is focused on the study of the protozoan parasite Leishmania, the aetiological agent of the important disease leishmaniasis. The disease is characterized as anthropozoonotic and is transmitted by the bite of sandflies. Leishmaniasis is endemic in Greece, whereby the prevalence in dogs, the local reservoir host, is up to 34%. In humans leishmaniasis has different clinical manifestations, including cutaneous, mucocutaneous and visceral, the latter being the most severe form of disease, lethal if left untreated. Leishmaniasis is considered a major health problem with more than 1.5 million infections occurring each year, whereas current chemotherapy has important limitations including high toxicity and  emergence of drug resistance. Thus, there is an urgent need to discover new drugs. For contributing to the fight against leishmaniasis the first research axis of the group investigates the pathogen and host signal transduction pathways, aiming to expand the drug-discovery platforms for the treatment of the disease.  In this context, we use molecular and cellular biology techniques and systems approaches, to focus on the druggable genome and more specifically on kinases of the host implicated in inflammation (i.e. GSK-3, DYRK) and on parasitic kinases implicated in parasite viability, virulence and differentiation (i.e. GSK-3, Aurora, DYRK). Aiming towards the discovery of new antiparasitic drugs we have developed target- and cell- based screening assays, and we evaluate the efficacy of selected drugs in vivo, in animal models of leishmaniasis. The second research axis involves epidemiological studies of strains isolated in South-Eastern Europe for understanding parasite factors that determine the clinical outcome of leishmaniasis and factors that contribute to drug resistance.

Α. Investigation of signal transduction pathways and of proteins involved in Leishmania cell-cycle progression, differentiation and virulence

To achieve our research objectives we use transgenic parasites, molecular and cellular biology techniques and systems approaches for investigating components and mechanisms of signal transduction pathways.

PUBLICATIONS

• Leishmania dual-specificity tyrosine-regulated kinase 1 (DYRK1) is required for sustaining Leishmania stationary phase phenotype.Rocha VPC, Dacher M, Young SA, Kolokousi F, Efstathiou A, Späth GF, Soares MBP, Smirlis D. Mol Microbiol. 2020 May;113(5):983-1002. doi: 10.1111/mmi.14464.
• The loss of virulence of histone H1 over-expressing Leishmania donovani parasites is directly associated with a reduction of HSP83 rate of translation. Alexandratos, J. Clos, M. Samiotaki, A. Efstathiou, G. Panayotou, K. Soteriadou, D. Smirlis. Mol. Microbiology. 2013 Jun;88(5):1015-31.
• Trypanosomatid apoptosis: ‘Apoptosis’ without the canonical regulators. D. Smirlis, K. Soteriadou. Virulence. 2011 May 1;2(3).
• Targeting essential pathways in trypanosomatids gives insights into protozoan mechanisms of cell death. D Smirlis, M. Duszenko, A. Jimenez-Ruiz, E. Scoulica, P. Bastien, N. Fasel, K. Soteriadou. Parasit Vectors. 2010 Nov 17;3:107. (Highly accessed)
• Leishmania donovani RAN-GTPase interacts at the nuclear rim with linker histone H1. D. Smirlis, H. Boleti, M. Gaitanou, M. Soto, K. Soteriadou. Biochem. J. 2009 Dec 10;424(3):367-74 9.
• Leishmania histone H1 over-expression delays parasite cell-cycle progression, parasites differentiation and reduces Leishmania infectivity in vivo. D. Smirlis, S. Bisti, E. Xingi, G. Konidou, M. Thiakaki, K. Soteriadou. Mol. Microbiology. 2006; 60(6): 1457-73

Β. Drug discovery efforts against leishmaniasis and trypanosomiasis and investigation of the mechanism of drug action

To achieve this objective we have developed medium throughput non-radioactive kinase assays for investigating the inhibitory potential of Leishmania kinases and we assess cell growth inhibition and mechanism of drug action (cell-cycle, mechanism of cell death). Finally we assess the toxicity of selected compounds and their in vivo efficacy in a murine model of leishmaniasis.

PUBLICATIONS

• Indirubin Analogues Inhibit Trypanosoma brucei Glycogen Synthase Kinase 3 Short and T. brucei Growth. Efstathiou A, Gaboriaud-Kolar N, Myrianthopoulos V, Vougogiannopoulou K, Subota I, Aicher S, Mikros E, Bastin P, Skaltsounis AL, Soteriadou K, Smirlis D. Antimicrob Agents Chemother. 2019 May 24;63(6):e02065-18. doi: 10.1128/AAC.02065-18.
• Indirubin analogues inhibit Typanosoma brucei Glycogen Synthase Kinase 3 short and T. brucei growth.. Efstathiou A, Gaboriaud-Kolar N, Myrianthopoulos V, Vougogiannopoulou K, Subota I, Aicher S, Mikros E, Bastin P, Skaltsounis AL, Soteriadou K, Smirlis D. Antimicrob Agents Chemother 2019 Mar 25. pii: AAC.02065-18. doi: 10.1128/AAC.02065-18. [Epub ahead of print]
• Indirubin derivatives are potent and selective anti-Trypanosoma cruzi Efstathiou A, Meira CS, Gaboriaud-Kolar N, Bastos TM, Rocha VPC, Vougogiannopoulou K, Skaltsounis AL, Smirlis D, Soares MBP. Virulence. 2018;9(1):1658-1668. doi: 10.1080/21505594.2018.1532242. PMID: 30387370
• Selection of molecular targets for drug development against trypanosomatids. D. Smirlis& M.B.P. Soares. In Proteins and Proteomics of Leishmania and Trypanosoma, Springer (Editor: dos Santos, A.L.S), Subcell Biochem. 2014;74:43-76. doi: 10.1007/978-94-007-7305-9_2. (Book Chapter)
• 5-Me-6-BIO targeting the leishmanial short form affects cell-cycle progression and induces apoptosis like death: exploitation of GSK-3 for treating leishmaniasis. E. Xingi, D. Smirlis, V. Myrianthopoulos, P. Magiatis, K. M. Grant, L. Meijer, E. Mikros, A. -L. Skaltsounis, K. Soteriadou. Int. J. Parasitol. 2009;39(12):1289-303
• In vitro activity of 10-deacetylbaccatin-III against Leshmania donovani promastigotes and intracellular amastigotes. K. Georgopoulou, D. Smirlis, S. Bisti, E. Xingi, L. Skaltsounis, K. Soteriadou. Planta Medica 2007 Aug;73(10):1081-8. Epub 2007 Aug 9

C. Epidemiological studies of strains isolated in South-Eastern Europe for understanding parasite factors that determine the clinical outcome of leishmaniasis and factors that contribute to drug resistance.

We use parasites MLMT typed that share the same genomic background to investigate distict phenotypes towards clinical outcome and drug resistance. By applying next generation sequencing or and other molecular biology approaches, we are trying to investigate on the mechanisms that contribute to the above phenotypes.

PUBLICATIONS

• Genetic diversity and structure in Leishmania infantum populations from south-eastern Europe revealed by microsatellite analysis. E. Gouzelou, C. Haralambous, M. Antoniou, V. Christodoulou, F. Martinković, T. Živičnjak , D. Smirlis, F. Pratlong, J.P. Dedet, Y. Özbel, S.Ö. Toz, W. Presber, G. Schönian, K. Soteriadou. Parasit Vectors. 2013 Dec 5;6:342.doi: 10.1186/1756-3305-6-342. (Highly accessed)

D.Host-pathogen interactions.

We investigate host and pathogen mechanisms that can modify virulence and pathogenicity

PUBLICATIONS

• SILAC-based quantitative proteomics reveals pleiotropic, phenotypic modulation in primary murine macrophages infected with the protozoan pathogen Leishmania donovani. Smirlis D, Dingli F, Pescher P, Prina E, Loew D, Rachidi N, Späth GF. J Proteomics. 2020 Feb 20;213:103617. doi: 10.1016/j.jprot.2019.103617.
• Osteopontin in the host response to Leishmania amazonensis. Giraud E, Rouault E, Fiette L, Colle JH, Smirlis D, Melanitou E. BMC Microbiol. 2019 Feb 8;19(1):32. doi: 10.1186/s12866-019-1404-z.

2016-2017: IKY: Scholarship of Excellence for post-doctoral studies Siemens, (Recipient: Dr Antonia Efstathiou)

2014-2017: International Pasteur Network: LeisHield: A new collaborative action to determine prevalence, anticipate emergence and assess urbanization of cutaneous and visceral leishmaniasis in LeishRIIP partner countries (Coordinator: Gerald Spath, PI: PI)

2013-2015: ACIP 2013 International Pasteur Network: “Evaluation of Leishmania DYRK family of kinases as molecular targets for the development of antileishmanial drugs” (Hellenic Pasteur Institute, IP Paris, IP Montevideo, FIOCRUZ) (Coordinator: Despina Smirlis)

2011-2015: FP7-PEOPLE-2010-IRSES: Exploring Chemical Biodiversity with Innovative Approaches for Fighting Chagas and Leishmaniasis. PIRCES-GA-2010-269301 (Coordinator: A.-L. Skaltsounis, PI: Ketty Soteriadou, Member, Co-PI: Despina Smirlis)

2012-2015: General Secretariat of Research and Technology KRIPIS: Infectious and neurodegenerative diseases of the 21st Century: From the study of basic mechanisms to translational research and development of cutting-edge technologies aiming towards prevention, diagnosis and treatment (WP leader), (WP Leader)

2011-2012: Foundation for Education and European Culture (IPEP) award: “Evaluation of the anti-leishmanial action of new indirubin analogues” (Recipient of Grant: Despina Smirlis)

TEACHING ACTIVITIES

Invited lectures/ Courses/ Seminars

1. Research seminar of the Biology Department of the National Kapodistrian University of Athens: The unusual lives of trypanosomatids, 19-12-2012.
2. UMR8638 CNRS-Université Paris Descartes, “Novel drug targets and drugs for the treatment of leishmaniasis, 19-7-2013.
3. Institute Pasteur Paris, Scientific Symposium of the Institut Pasteur International Network “Exploring the role of Leishmania infantumDYRK1 and Aurora kinases in the parasitic lifecycle for anti-leishmanial drug discovery”, 11-9-14.
4. Institut Pasteur, Tehran, Iran: International Workshop “Leishmania and leishmaniasis immunology, vaccine and drug discovery-therapy” Speaker: Recent approaches in anti-leishmanial drug discovery, 9-13 November 2016.

Theses/ Final Year Projects

2012-now: Currently co-supervising with Dr Milena Soares (FIOCRUZ, Bahia, Brazil) Mr. Vinicius Pinto Rocha

2017: Foteini Kolokousi Dissertation supervisor (Biology, NKUA) in the project entitled “Exploring the role of Leishmania infantum DYRK1 for antileishmanial drug discovery”

2016: Garyfallia Loulou Dissertation supervisor (Technical University of Athens) in project entitled “Phenotypic characterisation of Leishmania infantum parasites knocked out or over-expressing the Leishmania dual specificity tyrosine regulated kinase 1 (DYRK1), for the study of its biological role”

2010-2014: Co-supervisor of Antonia’s Efstathiou PhD thesis entitled (Biology, National Kapodistrian University of Athens) entitled «Study of molecules implicated in the development of Leishmania and T.brucei parasites»

2010-2013: Co-supervisor of Alexandros Alexandratos PhD thesis (Chemistry, University of Ioannina) “Study of molecules important for cell-cycle progression in the protozoan parasite Leishmania”

2009: Evangelos Nomikos BSc Final Year Project Supervisor (Technical University of Athens) in the final Year Project entitled “Expression and purification of a Leishmania RCC1 protein fragment”

2008: Irene’s Daoudadakis BSc Final Year Project Supervisor (Technical University of Athens) in a project entitled “Construction of targeted cassettes for the knockout of Leishmania protein Ran”

2007-2008: Anastasios Katopodis Final Year (Biology, National Kapodistrian University) Project Supervisor in a project entitled “Cloning and molecular characterisation of the Leishmania protein Ran”

Stavroula Gouzelou