Tutkimuksen ja opetuksen kuvaus
Present research project
PRINT-AID project, PhD in Drug Research (2017-present)
Supervised by Jari Yli-Kauhaluoma, Kirsi Savijoki and Adyary Fallarero, Faculty of Pharmacy, Division of Pharmaceutical Biosciences, Anti-infective research group, University of Helsinki, Finland
Title of thesis: Identification, mechanistic characterization and chemical optimization of repurposed anti-biofilms
Description: The doctoral project is part of the PRINT-AID network (Marie Sklodowska-Curie Actions, Innovative Training Networks, H2020-MSCA-ITN-2016) and contributes to the main goal of developing a new generation of personalized anti-infective 3D-printed medical devices. The objective of the project is to identify repurposed anti-biofilm compounds from libraries of investigational and approved drugs. Promising molecules will be structurally optimized in order to improve their activity and integrate them into a 3D-printed formulation. The mode of action of selected compounds will also be investigated.
- Optimization of an antimicrobial activity screening platform based on in vitro bacterial biofilms staining to a 384-well microplate format
- High-throughput evaluation of the preventive and destructive antimicrobial activity of libraries of compounds on bacterial biofilms using the optimized screening platform based on resazurin and crystal violet staining
- Characterization of the anti-biofilm activity using various techniques (ATP production, LIVE/DEAD staining and observation by microscopy, CFU count, MIC/MBIC determination, cytotoxicity, etc)
- Study of the quorum snesing inhibitory activity of promising compounds by quantification of violacein production using a recombinant Chromobacterium violaceum model
- Analysis of the physico-chemical properties of anti-biofilm compounds and structural optimization of the molecules using medicinal chemistry cycles and computational chemistry
- Mode of action studies of anti-biofilm compounds using various techniques (next-generation sequencing, inactivation/overexpression of target genes by production of mutants, confirmation of direct interaction (ELISA, ChIP), fluorescent microscopy (IF, PNA probe, recombinant plasmids), proteomic studies (footprint)
PhD in Drug Research (2017 - Ongoing), University of Helsinki, Finland
MSc in Microbiology-Immunology (2016), Laval University, Quebec City, Canada
BSc in Biomedical sciences (2014), Laval University, Quebec City, Canada
BSc in Biology (2014), Laval University, Quebec City, Canada
Previous Research Experience
MSc in Microbiology-Immunology project (2014-2016)
Supervised by Louis Flamand, Division of Infectious disease and immunity, CHU de Québec Research Center (CHUL), Laval University, Quebec City, Canada
Title of thesis: Study of the chromosomal integration of human herpesvirus 6 and impact of its infection on damages recognition at telomeres
Tasks and accomplishments:
- Analyzed viral and cellular genes expression at the RNA and protein levels in HHV-6 infected cells. Cell culture in laboratory containment level 2 (cell lines U2OS, HeLa, MOLT-3, HSB-2, HUVEC, MCF-7, GM847). Infection kinetics with HHV-6A and HHV-6B. Virus production and titration. Isolation from blood samples, culture and infection of monocytes.
- Demonstrated a difference in the shelterin complex RNA expression (TRF1, TRF2, TPP1) and in the TRF2 protein expression during HHV-6 infection. DNA and RNA extraction, RT-qPCR, Western Blot, ELISA, ChIP. Some experience in multicolor flow cytometry.
- Investigated expression and distribution of DNA damage response proteins (53BP1, yH2AX), TRF2 protein, viral proteins (IE1/2, p41) and telomeric sequences during HHV-6 infection. Molecular cloning. Analysis of RNA, DNA and protein sequences. Expression of recombinant proteins. Transduction (Lentiviruses). Immunofulorescence and FISH. Confical microscopy.
- Tested the effect of a telomerase inhibitor (BRACO-19) on chromosomal integration of HHV-6 in telomeres. Single-cell clones production (HeLa, U2OS, MCF-7). qPCR and ddPCR. Some ecperience with Surface Plasmon Resonance.
Research internship in Biomedical sciences (2014)
Supervised by Louis Flamand, Division of Infectious disease and immunity, CHUL, Quebec city, Canada
- Studied the human herpesvirus 6 pritein U94 and its interaction with telomeric protein TRF2 (DNA and RNA extraction, RT-qPCR, Western Blot, transfection, IF and confocal microscopy).
Research internship in biology (2013)
Supervised by Jean Charron, Centre of Cancer Research, Hôtel-Dieu de Québec, CHUQ, Quebec City, Canada
- Characterized a new animal model of cranio-facio-cutaneous syndrome: genetically modified mice (Mek1 y130c). DNA extraction and genotyping by Southern Blot. Sampling from mice (skull, brain, heart, embryos), fixation, embedding, cutting, staining and analysis of histological samples. (formation in biosecurity, radioprotection and work in an animal facility - mice)
Student assistant in the biobank project IRNPQEO (2012)
Laboratory of Emmanuel Bujold, Centre of perinatal research, CHUQ, Quebec City, Canada
- Participated in the collection, freezing and storing of human biological samples (placenta and umbilical biopsies, blood, etc) to create a biobank of mother-child paired samples.
French (native), English (proficient), Finnish (basic)
Gilbert-Girard, S., Gravel, A., Artusi, S., Richter, S. N., Wallaschek, N., Kaufer, B. B., Flamand, L., Stabilization of Telomere G-Quadruplexes Interferes with Human Herpesvirus 6A Chromosomal Integration. J Virol, 2017. 91(14). DOI: 10.1128/JVI.00402-17
Gravel, A., Dubuc, I., Wallaschek, N., Gilbert-Girard, S., Collin, V., Hall-Sedlak, R., Jerome, K. R., Mori, Y., Carbonneau, J., Boivin, G., Kaufer, B. B., Flamand, L., Cell Culture Systems To Study Human Herpesvirus 6A/B Chromosomal Integration. J Virol, 2017. 91(14). DOI: 10.1128/JVI.00437-17
Trempe, F., Gravel, A., Dubuc, I., Wallaschek, N., Collin, V., Gilbert-Girard, S., Morissette, G., Kaufer, B. B., Flamand, L., 2015. Characterization of human herpesvirus 6A/B U94 as ATPase, helicase, exonuclease and DNA-binding proteins. Nucleic Acids Res 43:6084-6098. DOI: 10.1093/nar/gkv503
- 317 Farmasia
Assess the cell viability of Staphylococcus aureus biofilms. PrestoBlue HS and alamarBlue HS reagents for microplate viability assays.Gilbert-Girard, S., Savijoki, K. & Fallarero, A., 2019, julkaisussa : BioProbes Journal of Cell Biology Applications.
Tutkimustuotos: Artikkelijulkaisu › Artikkeli › Ammatillinen