Χαραλαμπία Μπολέτη

Haralabia Boleti | Intracellular Parasitism

Senior Researcher
+ 30 210-6478879
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RESEARCH INTERESTS

i. Interaction of Leishmania protist pathogens with mammalian host cells

Single celled eukaryotic parasites of the Leishmania genus, belong to the Trypanosomatidae order of the Kinetoplastid class of protists. They are dimorphic, living as flagellated forms (promastigotes) in the gastrointestinal tract of their sandfly vector and as aflagellated forms (amastigotes) mainly inside modified phagolysosomes of mammalian host phagocytes.  Metacyclic infectious forms of Leishmania promastigotes are transmitted to the mammalian host upon a blood meal of the insect vector (sandflies Phlebotomus or Lutzomyia). The parasite establishes infection in macrophages by which it is carried in organs distant from the site of infection as the liver, spleen and bone marrow in the case of visceral Leishmaniasis. Survival and propagation of the parasite in the professional phagocytes is due to interference and subversion of host signaling pathways related to many aspects of cell physiology including the ones of intracellular trafficking and secretion, phagolysosome biogenesis and apoptosis. 

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Infection of mammals with anthroponotic or anthropozoonotic Leishmania spp, causes a wide spectrum of human cutaneous (CL) and visceral (VL) diseases with high socioeconomic burden known as Leishmaniases. Although several drugs against the different forms of Leishmaniasis exist, there is an urgent need to develop new ones due to severe side effects, high cost and the development of drug resistant parasite strains, given that no efficient vaccine against the different forms of Leismaniasis exists for humans.

Research in our lab focuses on the study of surface (i.e. ecto-enzymes) or secreted proteins of Leishmania donovani as potential virulence factors interacting with the host. Moreover, we study a L. donovani phosphatase (product of LDBPK_220120 genes (TriTrypDB)] which belongs to a Dual Specificity and PI phosphatase family named Atypical Lipid Phosphatases (ALPs) expressed only in microorganisms some of which are pathogenic (Beresford, N.J. et al. 2010) which renders it a potential candidate drug target. Along the same line, in an effort to discover new drug targets for development of anti-leishmania compounds we focused on parasite proteins involved either in phosphoinositide (PI) metabolism (i.e. PI phosphatases or kinases) or are PI effectors through their PI binding domain. PIs regulate vital cellular processes and thereby proteins interacting with them play essential roles in a cell’s life. The field of PI signaling and PI binding proteins in Leishmania is very poorly studied and our study highlights important molecules for further analysis in parasite’s life and in virulence.

Lastly, we use the nonpathogenic to humans Leishmania tarentolae (L. tarentolae) species as 1) system for recombinant eukaryotic protein expression (Biotechnological applications) and 2) in vitro system for screening  anti-leismanial compounds.

ii. Anti-cancer properties of natural products-Olive oil polyphenols

Olive oil consumption is associated with prevention of many cancer types. Its chemopreventive activity is attributed to olive oil polyphenols (OOPs) such as hydroxytyrosol and tyrosol, along with their secoiridoid derivatives oleuropein aglycon, ligstroside aglycon, oleacein, and oleocanthal. Certain of these compounds have been shown to inhibit cancer cell proliferation and/or induce apoptosis. However, issues concerning safety/toxicity of OOPs, their mechanism(s) of action or synergistic effects in OOPs’ combinations remain to be yet clarified.

 

RESEARCH PROGRAMS

Study of surface (i.e. ecto-enzymes) or secreted proteins of Leishmania donovani as potential virulence factors interacting with the host.

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We have characterized an L. donovani specific membrane bound acid ecto-phosphatase (LdMAcP) and showed that it contributes to parasitic virulence in vitro (As L. donovani secreted molecule we study a phosphoinositide (PI) binding protein, product of the LDBPK_352470.1.1 gene (TriTrypDB), first discovered in a secretome proteomics analysis of L. donovani’s promastigotes (Silverman JM et al. 2008). This protein is secreted by parasites and thereby constitutes a candidate factor for parasite-host interaction. We have shown (Tziouvara O. et al. and Boleti 2021, small talk 71st conference of the *HSBMB) that LdPIBPsnx has the structural characteristics of members of the SNX-BAR subfamily of Sorting Nexins (Marina Petsana, Diploma thesis, 2021) and that it is highly conserved along the Evolutionary tree of Eukaryotes. This is the first Sorting Nexin identified and characterized in Leishmania spp and is the homolog of the human SNX2 protein (O. Tziouvara et al. and Boleti, manuscript in preparation). Sorting nexins constitute a protein family highly conserved in evolution, indicating the importance of these proteins in the eukaryotic cell life. Sorting nexins have not been studied before in parasites of the Leishmania genus.

PUBLICATIONS

  • Papadaki A, Politou AS, Smirlis D, Kotini MP, Kourou K, Papamarcaki T, Boleti H. (2015). The Leishmania donovani histidine acid ecto-phosphatase LdMAcP: insight into its structure and function. Biochem J. 467(3):473-86 (PDF).

THESES

  • PhD, Amalia Papadaki, Medical School University of Ioannina, Defended on April 27, 2015
    Title: Study of the molecular mechanisms underlying Leishmania survival within the phagocytes of the mammalian host
  • MSc, Drosos Kourounis, University of Athens, MSc Program «Applications of Biology in Medicine”. March 2017
    Title:Cloning and molecular characterization of the secreted nexin LdPIΒPnex with a PI binding domain from the protozoan parasite Leishmania donovani
  • MSc, Alexis GACHET-IEREMIADIS, M1 Master 1 Health engineering / Management of biotechnology, Grenoble Alpes University (UGA) 2016/17. defended in June 2017
    Title: Expression in E. coli and purification of a Leishmania donovani phosphoinositide binding protein from the nexin family (LdPIBPnex)
  • Diploma, Marina Petsana (Bioinformatician, BSc) (Jul. 2020-Feb. 2021). Department of Informatics with applications in biomedicine, School of Sciences, University of Thessaly, Greece. Diploma thesis title: “Bioinformatics analysis of the Ld1 gene product of the protozoan parasite Leishmania donovani

CONFERENCE PRESENTATIONS

  • 71st Congress of the Hellenic Society of Biochemistry and Molecular Biology (EEBMB). 26-28 Nov. 2021, Athens, Greece. Short talk title: «LdPIBPsnx: The First Sorting Nexin in Leishmania. Authors: Tziouvara, M. V. Petsana, D. Kourounis, A. Papadaki, E. Basdra, G. G. Braliou, H. Boleti
  • 67oPanhellenic conference of the Hellenic Society of Biochemistry and Molecular Biology (EEBMB). Ioannina, (25-27 Nov. 2017) Poster (prize)Title: LdPIΒPnex, a secreted nexin-like protein from the protozoan parasite Leishmania donovani. Authors Kourounis, A. Papadaki, O. Tziouvara, H. Boleti (Poster)
  • Scientific Symposium of the Institut Pasteur International Network. Paris, France (10-13 Sept. 2014). Poster Title: The Leishmania membrane bound Histidine Acid Phosphatases (HAcPs) as putative virulence factors; Characterization of the LdMAcP, a Histidine Acid ecto-phosphatase from AuthorsΑ. Papadaki, Η. Boleti (Poster)
  • Scientific International Meeting of Young Researchers from RIIP, Institut Pasteur, Paris, France (10 Nov. 2011). Commented Poster (oral presentation). Title: Phosphoinositide involvement in Leishmania donovani phagocytosis by Raw 264,7 macrophages. AuthorsΑ. Papadaki, Η. Boleti (Poster)

Structural and functional characterization of specific L. donovani phosphatases as candidate anti-Leishmanial drug targets.

Main focus of our work is the study of the LdMacP (LdAcP3.1_AF149839.1) Histidine Acid ecto-phosphatase from L. donovani (Shakarian, A.M., et al.2002, Papadaki et al. 2015) and Leishmania proteins involved either in phosphoinositide (PI) metabolism (i.e. PI phosphatases or kinases) or are PI effectors through their PX PI binding domain. PIs regulate vital cellular processes and thereby proteins interacting with them play essential roles in a cell’s life.

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We study a P-Tyr/PI dual specificity phosphatase and have shown results indicating that this enzyme is expressed both in promastigotes and amastigotes and may play a role in parasite stage differentiation (Papadaki et al and Boleti, Front Cell Infect Microbiol, 2021). We have generated dominant negative mutants and plan to knock out the protein to explore its importance in the parasite’ life or virulence. This PI Phosphatase (product of the  LDBPK_220120 gene (TriTrypDB)] belongs to a family of  Dual Specificity and PI phosphatases named Atypical Lipid Phosphatases (ALPs) expressed only in microorganisms some of which are pathogenic (i.e Mycobacterium tuberculosis; Beresford, N.J. et al. 2010). It therefore represents attractive candidate as potential drug target.

Moreover, by a systematic review and in silico analysis of protein sequences containing PX domain in Kinetoplastea protists we discovered several proteins in Leishmania spp. which contain an evolutionary conserved PX PI-binding domain, and we classified them in four distinct groups on the basis of their overall domain architecture. This study highlighted several new candidates of PI-interacting proteins in Trypanosomatida (Petsana et al. Boleti & Braliou, ISCB-Africa ASBCB Conference on Bioinformatics 2021; 21st HSCBB international hybrid conference of the Greek society of computational biology and bioinformatics, 2021). The field of PI signaling and PI binding proteins in Leishmania is very poorly studied and our study highlights important molecules for further analysis in parasite’s life and in virulence.

PUBLICATIONS

  • Papadaki A, Politou AS, Smirlis D, Kotini MP, Kourou K, Papamarcaki T, Boleti H. (2015). The Leishmania donovanihistidine acid ecto-phosphatase LdMAcP: insight into its structure and function. Biochem J. 467(3):473-86. (PDF)
  • Papadaki A., Tziouvara O., Kotopouli A., Koumarianou P., Doukas A., Rios P., Tardieux I., Köhn M., Boleti H. (2021). The Leishmania donovani1 gene encodes for an atypical Dual Specificity Lipid-like phosphatase expressed in promastigotes and amastigotes; Substrate specificity, intracellular localizations and putative role(s). Frontiers in Cellular and Infection Microbiology, section Parasite and Host, 11: 85. (PDF).

THESES

  • PhD, Amalia Papadaki, Medical School University of Ioannina, Defended on April 27, 2015. Title: Study of the molecular mechanisms underlying Leishmania survival within the phagocytes of the mammalian host
  • MSc, Anastasia Kotopouli, ΕΚΠΑ, Chemistry Department, «Biochemistry» Defended in Feb. 2014. Title:Cloning, molecular and functional characterization of the putative PI phosphatase, LdPIP_22, of the parasitic protozoan  Leishmania donovani
  • ΜSc, Mathias Richard, Faculté de Sciences, University of Montpellier, MSc BioPMBB-IMH, Defence in June 2015. Title: Molecular Biology of the protozoan parasites Leishmania donovani: Cloning of an  infantum putative phosphatase into plasmid vectors for expression of His-tagged or mCherry-tagged protein in bacteria, mammalian and Leishmania cells.
  • MSc, Olivia Paraskevi Tziouvara, Biology Dept, U. of Athens (MSc program Applications of Biology in Medicine) Defence in October 2017. Title:Molecular and functional characterization of the Tyrosine and phosphoinositide phosphatase LdTyrPIP_22, from the protozoan parasite Leishmania donovani.

CONFERENCE PRESENTATIONS

  • ΕΕΒΜΒ 2021, «Molecular basis of infections» (Virtual congress), 21-26 May 2021. 5 min video poster: The Leishmania donovani Tyrosine and PI Phosphatase Ld  Authors: O. Tziouvara, P. Koumarianou, H. Boleti
  • ISCB-Africa ASBCB Conference on Bioinformatics 2021 (Virtual) Poster: «Phylogenetic and structural analysis of the PX domain containing PhosphoInositide (PI) binding proteins in Kinetoplastida». Authors: Marina V. Petsana, Ahmed F. Roumia, P. G. Bagos, Boleti & G. Braliou. (PDF)
  • 66thPanhellenic conference of the Hellenic Society of Biochemistry and Molecular Biology (ΕΕΒΜΒ) Athens (11-13 Dec. 2015). Oral presentation. Title: Biochemical characterization and subcellular localization of the tyrosine and phosphoinositide dual specificity phosphatase LdPIP_22, a potential drug target from Leishmania donovaniAuthors Papadaki, A. Kotopouli, A. Doukas, P. Rios, O. Tziouvara, M. Köhn, H. Boleti (PDF)
  • Cost Action CM0801, 4th Annual Meeting. Crete (Sept. 19-21 2012). Poster Title: Study of specific Leishmania Atypical Lipid Phosphatases (ALPs) as putative virulence factors for parasite survival in host phagocytes and as potential drug targets.Authors: Kotopouli, A. Papadaki, A. Doukas , K. Papadakos , D.Sgouras, N. Galanopoulou, H. Boleti. (PDF)
  • 62οPanhellenic conference of the Hellenic Society of Biochemistry and Molecular Biology (ΕΕΒΜΒ) Athens (9-11 Dec. 2011). Poster Title: Study of specific Leishmania histidine acid phosphatases as putative virulence factors for parasite survival in host phagocytes and as potential drug targets. Authors: Papadaki Α., Smirlis D., Kourou, K., Politou A.S., Boleti Η. (PDF)

Validation of the Leishmania tarentolae (L. tarentolae) organism (nonpathogenic to humans) as 1) system for recombinant eukaryotic protein expression (Biotechnological applications) and 2)  in vitro system for screening anti-leismanial compounds.

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We have successfully expressed in L. tarentolae promastigotes a mammalian membrane transporter (Doukas et al. 2019), the human PRL3 phosphatase (IKYDA program, collaboration with Prof. Dr Μaja Köhn, Institute of Biology III, Group: Integrative Signaling Studies, University of Freiburg) and the LdTyrPIP_22 L. donovani PI phosphatase (Papadaki et al. 2021; Dimotsantos G., MSc thesis 2021). For the screening of candidate anti-leismanial drugs we generated a genetically modified L. tarentolae strain expressing the mcherry (Doukas A. MSc thesis 2015). With this technology our group is open to collaborations on eukaryotic protein expression or screening of compounds with anti-leishmanial potential.

PUBLICATIONS

Doukas A, Karena E, Botou M, Papakostas K, Papadaki A, Tziouvara O, Xingi E, Frillingos S, Boleti H. (2019). Heterologous expression of the mammalian sodium-nucleobase transporter rSNBT1 in Leishmania tarentolae. Biochim Biophys Acta Biomembr. 1861(9):1546-1557. doi:10.1016/j.bbamem.2019.07.001

THESES

  • MSc, Anargyros DOUKAS, Biology Departments, University of Athens, MSc program in «Microbial BIotechnology». Defence in Jan. 2015. Title:Construction and characterization of transgenic διαγονιδιακών Leishmania tarentolae strains for heterologous expression of the red fluorescent protein mCherry and of the nucleobase transporter rSNBT1 (from Rattus norvegicus) (PDF)
  • ΜSc, Georgios Dimotsantos, Biology Dept, U. of Athens, MSc program “Microbial Biotechnology”. Defence  in Dec. 2021. Title: The microorganism Leishmania tarentolae as a tool for the expression and isolation of recombinant proteins from homologous microorganisms in order to perform their biochemical and functional characterization.

CONFERENCE PRESENTATIONS

  • 2nd International Caparica Conference on Leishmania 2020, Caparica, Portugal 26th- 28th Oct. Invited oral presentation: Leishmania tarentolae as a model organism for heterologous expression of mammalian multi spanning membrane transporters. Pros and cons. Authors: H. Boleti, A. Doukas, S. Friligos
  • 64thPanhellenic conference of the Hellenic Society of Biochemistry and Molecular Biology (ΕΕΒΜΒ), Athens (6-9 Dec. 2013). Poster Title: Heterologous expression of the mammalian nucleobase transporter rSNBT1 in the LEXSY Leishmania tarentolae protein expression system. Authors Doukas A., Papakostas K., Papadaki A., Frillingos S., Boleti H. (PDF)

Anti-cancer properties of natural products-Olive oil polyphenols

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In collaboration with Prof. P. Magiatis (Section of Pharmakognosy and Natural Products, School of Pharmacy, National Kapodistrian University of Athens) we have initiated a systematic study on the anti-proliferative/cytotoxic effects of six OOPs (some of them never studied before) and of specific polyphenols’ combinations and extracts on multiple cell-culture based cancer models (work funded by a Greek Government/EU grant). This study is providing important results for further in vivo studies using cancer animal models (Papakonstantinou A., Koumarianou, P. et al. 2021 in preparation)

THESES

  • PhD, Katerina Papakonstantinou, Pharmacy department, University of Athens. Title: Development of new analytical techniques for the bioactive phenols of olive and study of their activity in humans “. Start date:  June 2018- ongoing

CONFERENCE PRESENTATIONS

  • 20th International congress of the International Society of Ethnopharmacology, Virtual congress 18-20 April 2021.  i. Poster: Evaluation of the antiproliferative/cytotoxic action of olive oil polyphenols on human breast cancer. Authors:  A. Papakonstantinou, P. Koumarianou, E. Melliou, P. Magiatis, H. Boleti.  ii.Poster: Evaluation of antiproliferative/cytotoxic action of olive oil polyphenols on human melanoma cancer. Authors: P. Koumarianou, A. Papakonstantinou, E. Melliou, P. Magiatis, H. Boleti

CURRENT GROUP MEMBERS

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Olympia-Paraskevi Tziouvara

Biologist (BSc)
MSc Student (Applications of Biology to Medicine)
Sep. 2015- May 2017
PhD Student 2018-
Email: .img@.img

Παπακωνσταντινου_tiny5-2

Katerina Papakonstantinou

Pharmasist (BSc, MSc)
PhD Student,
Pharmacy School, Univesrity of Athens
Email: .img@.img

Μαρίνα Πετσανά95

Marina Petsana

Bioinformatician (BSc), MSc student Dep. of Informatics with applications in Biomedicine, University of Thessaly, Greece,
July-Jan 2020 undergraduate diploma thesis, MSc thesis Feb. 2021-

agronomist - specialty biotechnology

Tasakou Maria

Agronomist – Specialty Biotechnology
MSc Student, “Imfectious diseases – international medicine Program: from the laboratory to clinical practice”, Democretian University, Thrace, Greece , Dec.1 2021- .

OLDER GROUP MEMBERS / COLLABORATORS

Δημοτσάντος80

Giorgos Dimotsantos

Biologist/Biotechnologist (BSc)
MSc student, Sep. 2018- Dec. 2021
Βiology Department “Microbial Biotechnology”, University of Athens

Danniel Carrasco

Daniel Carrasco

Molecular Biologist (BSc, MSc)
PhD student (Calmette Yersin PhD fellow), Oct.1, 2020 – Aug. 2021
University of Alcala, Madrid
Πετρίνα80

Petrina Koumarianou

Biologist/Cell Biologist (ΒSc, MSc, PhD)
Post-doctoral fellow Sep. 2018
Email: .img@.img

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Panagiotis Samboutzis

Biologist, University of Patras,
Department of Environmental and Natural Resources Management
PhD Fellow (Collaborator from Dr Bourtzis’ lab) 2007-2008

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Maria Parakevi Kotini

Biologist Diploma thesis, Μarch 2008-Νοv. 2009

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Anastasia Kotopouli

Chemist ΜSc Student (Biochemistry), Sep. 2012 – Jan. 2014

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Dr Rositsa Tropcheva

Food Biotechnologist (PhD)
Assistant Professor, “St Kliment Ohridski” University of Sofia, Bulgaria, Research Fellow, June 2014- July 2014 (Institut Pasteur International Network Chalmette Yersin fellowship)

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Anargyros Doukas

Chemist
Technical Assistant & ΜSc Student (Microbial Biotechnology), June 2012- Jan. 2015

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Mathias Richard

Faculté de Sciences, U. of Montpellier,
MSc Student – BioPMBB-IMH, Feb. 2015- Jul. 2015

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Amalia Papadaki

Veterinarian, ΜSc,
PhD Fellow, Jun 2009- May 2015
Post doctoral Fellow, Jun 2015-Jan. 2016

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Drosos Koyrounis

Biologist (BSc)
MSc Student (Applications of Biology to Medicine),
Sep. 2015-Mar. 2017

hakan sukur

Hakan Nizam

5th year undergraduate student, Biotechnology Department, Agricultural University of Athens
Practical training, Oct 2020.-January 2021

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Alexis Gachet-Ieremiadis

ΜSc Internship, Biotechnologist (BSc)
April 2017- June 2017

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Maria Boulougari

4rth year undergraduate student,
Biotechnology Dep., Agricultural U. of Athens
Summer student training, Jul. – Sep. 2019

SELECTED PUBLICATIONS

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Papadaki A., Tziouvara O., Kotopouli A., Koumarianou P., Doukas A., Rios P., Tardieux I., Köhn M., Boleti H. (2021). The Leishmania donovani LDBPK_220120.1 gene encodes for an atypical Dual Specificity Lipid-like phosphatase expressed in promastigotes and amastigotes; Substrate specificity, intracellular localizations and putative role(s). Frontiers in Cellular and Infection Microbiology, section Parasite and Host. 11:591868.doi: 10.3389/fcimb.2021.591868. (IF 4.123)

2021
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Papadaki A. and Boleti H. (2019). Measurement of Acid Ecto-phosphatase Activity in Live Leishmania donovani Parasites. BioProtocols (A peer-reviewed protocol journal. No publication fee; no access fee) following journals’ invitation. Vol 9 (19). DOI:10.21769/BioProtoc.3384

Doukas A, Karena E, Botou M, Papakostas K, Papadaki A, Tziouvara O, Xingi E, Frillingos S, Boleti H. (2019). Heterologous expression of the mammalian sodium-nucleobase transporter rSNBT1 in Leishmania tarentolae. Biochim Biophys Acta Biomembr. 1861(9):1546-1557.

Braliou GG, Kontou PI, Boleti H, Bagos PG. (2019). Susceptibility to leishmaniasis is affected by host SLC11A1 gene polymorphisms: a systematic review and meta-analysis. Parasitol Res. 118(8):2329-2342. Review.

Frakolaki E, Kalliampakou KI, Kaimou P, Moraiti M, Kolaitis N, Boleti H, Koskinas J, Vassilacopoulou D, Vassilaki N. (2019). Emerging Role of l-Dopa Decarboxylase in Flaviviridae Virus Infections. Cells 8(8). ( PDF )

2019
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Papadaki A, Politou AS, Smirlis D, Kotini MP, Kourou K, Papamarcaki T, Boleti H. (2015). The Leishmania donovani histidine acid ecto-phosphatase LdMAcP: insight into its structure and function. Biochem J. 467(3):473-86. ( PDF )

2015
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Athanasopoulos A, Boleti H, Scazzocchio C, Sophianopoulou V. (2013). Eisosome distribution and localization in the meiotic progeny of Aspergillus nidulans. Fungal Genet Biol. 53:84-96. ( PDF )

2013
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Boleti H., Smirlis D., Dalagiorgou G., Meurs E., Christoforidis S., & Mavromara.P. (2010). ER targeting and retention of the HCV NS4B protein relies on the concerted action of multiple structural features including its transmembrane domains. Mol. Memb. Biol. 27(1):50-74. ( PDF )

2010
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Smirlis D, Boleti H, Gaitanou M, Soto M, Soteriadou K. (2009). Leishmania donovani RAN-GTPase interacts at the nuclear rim with linker histone H1. Biochem J. 10;424(3):367-74. ( PDF )

2009
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Vassilaki N, Boleti H, Mavromara P.(2008).Expression studies of the HCV-1a core+1 open reading frame in mammalian cells. Virus Res. 133(2):123-35. ( PDF )

2008
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Vassilakι Ν, Boleti Η. and Mavromara P. (2007). Expression studies of the core+1 protein of the hepatitis C virus 1a in mammalian cells: the influence of the core protein and proteasomes on the intracellular levels of core+1. FEBS J. 274(16):4057-74. ( PDF )

2007
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Boleti H. & Robotis, J.F (2004). Viral Hepatitis. xPharm 1.0, edited by S.J. Enna dn David, B. Bylund, Published by, Inc. Elsevier. Reviews-Electronic publications ( PDF )

Boleti H. (2004). Chlamydia Infections. xPharm 1.0, edited by S.J. Enna dn David, B. Bylund, Published by, Inc. Elsevier. ( Link )

2004
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Ibrahim-Granet, O., Philippe,B., Boleti, H., Boisvieux-Ulrich, E., Prévost, MC., Grenet, D., Stern, M., and JP Latgé. (2003). Phagocytosis and intracellular fate of Aspergillus fumigatus conidia in alveolar macrophages. Infection and Immunity, 71(2):891-903. ( PDF )

2003
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Boleti, H., Ojcius, D. and Dautry-Varsat, A. (2000). Fluorescent labelling of intra-cellular bacteria in living host cells. J. Microbiological Methods, 1;40 (3): 265-274. ( PDF )

Poupel, O., Boleti H., Axisa, S., and Tardieux, I. (2000). Toxfilin, a novel actin binding protein from Toxoplasma gondii, sequesters actin monomers and caps actin filaments. J. Mol. Biol. of the Cell, 11(1): 355-68. ( PDF )

2000
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Boleti, H., Benmerah, A., Ojcius, D., Cerf-Bensussan,N., Dautry-Varsat, A. (1999). Chlamydia infection of epithelial cells expressing dynamin and Eps15 mutants:clathrin-independent entry into cells and dynamin-dependent productive growth. J. of Cell Science, 112:1487-1496. ( PDF )

1999

ΕDUCATION

PhD theses

Ongoing

  • Katherina Papakonstantinou (Pharmasist BSc, ΜSc), Pharmacy School, University of Athens. “Development of new Analytical techniques for the Bioactive phenols of Olives and study of their activity in humans”. Ιnitiation June
  • Olympia Tziouvara (Biologist, BSc ΜSc). Medical School, National Kapodistrian University of Athens.  “PI phosphatases and PI binding proteins of the Leishmania protozoan parasite: Characterization of two representative molecules and study of their role in the parasite life cycle and in the infection of the mammalian host”. Initiation: January 2019

Defended

Amalia Papadaki, Medical School, U. of Ioannina, defended in April 2015
Title: Study of the molecular mechanisms underlying Leishmania spp. survival within the phagocytes of the mammalian host (PDF)

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MSc theses

Anastasia Kotopouli, Chemistry Dep., U. of Athens (MSc Program Biochemistry), defended in February 2014
Title: Cloning, molecular and functional characterization of the putative phosphoinositide phosphatase LdPIP22 from the parasitic protozoan Leishmania donovani (PDF) 

Anargyros Doukas, Biology Dept, U. of Athens (MSc program Microbial Biotechnology), defended in January 2015
Title: Construction and characterization of transgenic strains of the protozoan Leishmania tarentolae for heterologous expression of the red fluorescent protein mCherry and nucleobase transporter rSNBT1 from Rattus norvegicus. (PDF) 

Mathias Richard, Faculté de Sciences, U. of Montpellier, MSc BioPMBB-IMH, defended in June 2015
Title: Molecular Biology of the protozoan parasites Leishmania donovani: Cloning of an L. infantum putative phosphatase into plasmid vectors for expression of His-tagged or mCherry-tagged protein in bacteria, mammalian and Leishmania cells. (PDF) 

Olivia Paraskevi Tziouvara, Biology Dept, U. of Athens (MSc program Applications of Biology in Medicine) to be defended at the end of 2016/beginning of 2017
Title: Molecular and functional characterization of the Tyrosine and phosphoinositide phosphatase LdTyrPIP_22, from the protozoan parasite Leishmania donovani. (PDF)

Drosos Kourounis, Biology Dept, U. of Athens (MSc program Applications of Biology in Medicine) to be defended at the end of 2016/beginning of 2017
Title: Cloning and molecular characterization of the secreted Phosphoinositide Binding nexin-like protein LdPIΒPnex from the protozoan Leishmania donovani (PDF)

MSc Alexis Gachet-Ieremiadis, M1 Master 1 Health engineering / Management of biotechnology, Grenoble Alpes University (UGA)  2016/17. defended in June 2017
Title: Expression in E.coli and purification of a Leishmania donovani phosphoinositide binding protein from the nexin family (LdPIBPnex)
 (
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ΜSc Georgios Dimotsantos, Biology Department, National Kapodistrian University of Athens (MSc program “Microbial Biotechnology”). Defense in Dec. 2021. Title: The microorganism Leishmania tarentolae as a tool for the expression and isolation of recombinant proteins from homologous microorganisms in order to perform their biochemical and functional characterization.

Diploma thesis

Μarina Petsana, Diploma thesis, Dep. of Bioinformatics with applications in Biomedical Sciences, University of Thessaly. Defended in Feb. 2021
Title: Bioinformatics Analysis of the protein encoded by the gene LdBK_352470.1 of the Leishmania donovani protozoan parasite”

Maria Kotini, Diploma thesis, Biology Dept, of Athens, Defended in Nov. 2009.
Title: Cosntruction and characterization of trasngenic parasites Leishmania donovani expressing the red fluorescent protein mRFP1and of anti-mRFP1 polyclonal antibody.

Teaching in graduate programs(2010-2020)

2020– Lecture: The flagellated protozoan parasite Leishmania tarentolae as a model for expression of recombinant eukaryotic proteins and for the study of cilia biogenesis (ciliogenesis). MSc program «Functional analysis of genes: From the design to the Live systems (ΛΣΕΣ)*», Medical School, University of Ioannina

2018 – Lecture “ The protozoan flagellated parasite Leishmania as a model for expression of recombinant eukaryotic proteins and for the study of ciliogenesis”. MSc program “Microbial Biotechnology” Biology Department, University of Athens.

2011-2017-Lectures «Leishmania tarantolae, a eucaryotic cell system with important biotechnological applications». MSc program «MICROBIAL BIOTECHNOLOGY», Department of Biology, University of Athens

2015 – Lectures «Host-Cell mechanisms that become targets of intracellular pathogens» MSc program «APPLICATIONS OF BIOLOGY IN MEDICINE» Biology Department, University of Athens

2011-2014 – Lectures «Study of Cellular Organization with modern Fluorescene Microscopy techniques» MSc program “BIOCHEMISTRY”, Department of Chemistry, University of Athens

Organization of International and courses (2006-2016)

2007-2008

Cutting edge technologies and their contribution to the Biomedical Sciences
3-day practical courses for Biology Teachers of Secondary School education. An activity for diffusion of Advanced Scientific Knowledge to selected groups of society. ( Poster )

2013

Digital image processing/analysis tools in Light Microscopy: From the basics and beyond. Institut Pasteur Network  Regional course 2013 ( Poster )

2016

Cell Biology and infection: Digital Image Processing/Analysis Tools for Quantitative Light Microscopy Imaging. Institut Pasteur International Network course 2016 ( Poster )

FUNDING

2020

CALMETTE YERSIN PhD fellowship Institut Pasteur International Network (IPIN)
PhD fellowship to Daniel Carrasco Navarro
Project title: “ Identification of molecular partners and functional characterization of a secreted Sorting Nexin (SNX) Protein from the protozoan parasite L. donovani; the first SNX to be studied from Leishmania spp.”
Grant initiation date: October 2020; Grant duration: 36 months
Budget: 60,000€

2019

Ministry of Development and Investment/grant Code ΕΒΒΜ103.
“Supporting researchers with emphasis on young researchers – B cycle”-
“Human Resources Development, Education and Lifelong Learning” on Priority Axis 6, co-funded by the European Social Fund (ESF).
Project title: 
In vitro study of the anticancer properties of polyphenols from greek olive oil.
Grant initiation date: 
End of 2019 Grant duration: 15 months
Budget: 
45,500€

H.F.R.I (Hellenic Foundation for Research and Innovation)
PhD grant (Olympia Tziouvara)
Project title: “PI phosphatases and PI binding proteins of the Leishmania protozoan parasite: Characterization of two representative molecules and study of their role in the parasite life cycle and in the infection of the mammalian host”
Grant initiation date: October 2019; Grant duration: 30 months
Budget: 27,000€

2018

ΒΙΟΙΜAGING GR“Hellenic Research Infrastructure for the Imaging and Observation of Fundamental Processes in Biology and Medicine” (MIS 5002755) which is implemented under the Action “Reinforcement of the Research and Innovation Infrastructure”, funded by the Operational Programme “Competitiveness, Entrepreneurship and Innovation” (NSRF 2014-2020) and co-financed by Greece and the European Union (European Regional Development Fund). Responsible for 2WPs of the HPIs participation in the Infrastructure Network.

«INSPIRED: The National Research Infrastructure on Integrated Structural Biology, Drug Screening Efforts and Drug target functional characterization» MIS code 5002550 WP.2.1, WP 2.2

2017

KRIPIS II (2017-2019): DEVELOPMENT GRANTS FOR RESEARCH INSTITUTIONS/Greek General Secretariat for Research and Technology
Project title: “Infectious, autoimmune and neurodegenerative diseases: study of the pathogenetic mechanisms and development of diagnostic, prognostic and therapeutic approaches” Subproject ΠΕ 3.3: Structural and functional characterization of the Tyrosine and Phosphoinositide phosphatase LdTyrPIP_22 of the protozoan parasite Leishmania donovani (Μ4-Μ24) with aim to point out  new chemotherapeutical targets from antileismanial drugs (Coordinator Hellenic Pasteur Institute).
Budget for ‘Intracellular parasitism team22,900 €

2014

IKYDA (2014-2015) – PROGRAM of PROMOTION OF EXCHANGE AND SCIENTIFIC COOPERATION GREECE – GERMANY. Project title: LeishPhospho: Molecular characterization of specific phosphatases from the intracellular protozoan parasite Leishmania donovani and exploration of their potential as drug targets. Scientific Responsible of Greek team: Dr Haralabia Boleti, Scientific Responsible of the German teamDr Μaja Köhn
Budget:10,000 €

Sponsorship by Abbvie Pharmaceutical to the research team of Intracellular parasitism for research on Leishmaniasis. Scientific Responsible: Dr Haralabia Boleti. Budget: 5,000 €

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2013

PLATON-BILATERAL S & T COOPERATION GREECE –FRANCE (2013-2015).
Project title: LeishPhosphoToxMolecular and functional characterization of specific phosphatases from the anthropozoonotic intracellular protozoan parasites Leishmania donovani and Toxoplasma gondii and investigation of their role in the parasite interaction with host cells; Exploration of their potential as drug targets.
Scientific Responsible of Greek team: Dr Haralabia Boleti,
Scientific Responsible of  French teamDr Isabelle Tardieux
Budget: 30,000 €

ΚΡΗΠΙΣ Ι (2013-2015): GREEK GENERAL SECREATARIAT of RESEARCH and TECHNOLOGY
Project title: Infectious and Neurodegenerative diseases in the 21st century. From the study of basic mechanisms to development of  translational research and cutting edge technologies with aim to improve preventive diagnosis  and treatment. Subproject EE2.5: Survival mechanisms of the L. donovani parasites in the macrophage host cell and identification of virulence factors.  Subproject EE 4.5:  Identification, evaluation and characterization of candidate molecular targets for the development of anti-leismanial drugs. (coordinations: Hellenic Pasteur Insitute).
Budget for Intracellular parasitism group: 42,900 €

2012

THALES (2012-2015): GREEK MINISTRY OF EDUCATION
Project title: EVOTRANS-Membrane Transport: Sturcture-Function relationships and Evolution.  Project coordiantor: E. Frilliggos. University of Ioannina. Subproject Π.Ε.1-4 scientific responsible: H. Boleti
Budget for Intracellular Parasitism group: 40,000 €

2011

Sponsorship by the pharmaceutical company Genesis PharmaProject titel: Study of the molecular mechanisms by which the protozoan parasite Leishmania donovani infects humans and domestic animals to cause the disease of Kala azar. Scientific responsible: Dr Haralabia Boleti
Budget: 15.000 €

HONORS

2016

Poster award at the 67ο Panhellenic conference of the Hellenic Society of Biochemistry and Molecular Biology, Ioannina, (25-27 Nov. 2017)Title: LdPIΒPnex, a secreted nexin-like protein from the protozoan parasite Leishmania donovani. AuthorsDrosos Kourounis, Amalia Papadaki, Olympia Tziouvara, Haralabia Boleti, Intracellular parasitism group, Hellenic Pasteur Institute

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