Engineering yeast cells for drug discovery

• Grant number: 15/03553-6
• Support type: Research Grants - Young Investigators Grants
• Duration: October 01, 2015 - December 31, 2021
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• Principal researcher: Elizabeth Bilsland
• Grantee: Elizabeth Bilsland
• Home Institution: Instituto de Biologia (IB). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Associated scholarship(s): 19/14146-3 - Engineering gene circuits in Saccharomyces cerevisiae, BP.MS; 19/17876-2 - New tools for synthetic biology in Saccharomyces cerevisiae, BP.IC; 19/15689-0 - Engineering gene circuits in Saccharomyces cerevisiae, BP.MS; + associated scholarships 18/25446-5 - Identification of pairs of transporters responsible for the import of xenobiotics through the plasma membrane of Saccharomyces cerevisiae, BP.IC; 18/05328-8 - Cloning and characterization of 10 human blood-brain-barrier transporters, BP.IC; 17/01986-8 - Determination of substrate specificity of plasma membrane transporters from Saccharomyces cerevisiae and Homo sapiens, BP.DR; 15/19103-0 - ENGINEERING YEAST CELLS FOR DRUG DISCOVERY, BP.JP - associated scholarships

Abstract

Drugs directed to the central nervous system frequently fail in clinical trials because of their poor transport into the brain. This is due to the inefficient import of drugs though the blood brain barrier. In this project, we aim to use the yeast Saccharomyces cerevisiae as a platform for studying the transport of chemicals into and out of cells. We will construct yeast strains with deletions of pairs of genes expressing of each of the 121 non-essential plasma membrane transporters, and use these in high-throughput screens to identify groups of proteins with overlapping substrate specificities as well as to define chemical groups preferentially using that particular import route. We will construct yeast strains where the native ergosterol synthesis pathway is replaced for a synthetic human cholesterol synthesis pathway. We have promising preliminary results indicating that strains with cholesterol in their plasma membranes have the ability to efficiently incorporate human trans-membrane proteins. Therefore, strains with a humanized plasma membrane can be hosts for the heterologous expression of blood-brain barrier transporters. Thus, they will be used in screening to identify chemical groups preferably imported by each route, facilitating efficient design of drugs directed to the central nervous system. The humanized strains may also be used in the search for inhibitors of cholesterol synthesis, and for large-scale expression of trans-membrane proteins for structural and functional studies. (AU)

Scientific publications

(7)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

ALMEIDA, LUDIMILA DIAS; SILVA, ALI SALIM FARAJ; MOTA, DANIEL CALIXTO; VASCONCELOS, ADRIELLE AYUMI; CAMARGO, ANTONIO PEDRO; PIRES, GABRIEL SILVA; FURLAN, MONIQUE; FREIRE, HELENA MARTINS RIBEIRO DA CUNHA; KLIPPEL, ANGELICA HOLLUNDER; SILVA, SUELEN FERNANDES; ZANELLI, CLESLEI FERNANDO; CARAZZOLLE, MARCELO FALSARELLA; OLIVER, STEPHEN G.; BILSLAND, ELIZABETH. Yeast Double Transporter Gene Deletion Library for Identification of Xenobiotic Carriers in Low or High Throughput. MBIO, v. 12, n. 6 NOV-DEC 2021. Web of Science Citations: 0.

ADVANCES IN PROTEIN CHEMISTRY AND STRUCTURAL BIOLOGY, VOL 124: PROTEIN KINASES IN DRUG DISCOVERY. Web of Science Citations: 0.

SILVA, SUELEN FERNANDES; KLIPPEL, ANGELICA HOLLUNDER; RAMOS, PRISCILA ZONZINI; SANTIAGO, ANDREE DA SILVA; VALENTINI, SANDRO ROBERTO; BENGTSON, MARIO HENRIQUE; MASSIRER, KATLIN BRAUER; BILSLAND, ELIZABETH; COUNAGO, RAFAEL MIGUEZ; ZANELLI, CLESLEI FERNANDO. Structural features and development of an assay platform of the parasite target deoxyhypusine synthase of Brugia malayi and Leishmania major. PLoS Neglected Tropical Diseases, v. 14, n. 10 OCT 2020. Web of Science Citations: 1.

FERREIRA, LETICIA TIBURCIO; RODRIGUES, JULIANA; CASSIANO, GUSTAVO CAPATTI; TAVELLA, TATYANA ALMEIDA; PERALIS TOMAZ, KAIRA CRISTINA; BAIA-DA-SILVA, DJANE CLARYS; SOUZA, MACEJANE FERREIRA; DO NASCIMENTO LIMA, MARILIA NUNES; MOTTIN, MELINA; ALMEIDA, LUDIMILA DIAS; CALIT, JULIANA; SILVA DE BARROS PUCA, MARIA CAROLINA; MELO, GISELY CARDOSO; BARGIERI, DANIEL YOUSSEF; PINTO LOPES, STEFANIE COSTA; GUIMARAES LACERDA, MARCUS VINICIUS; BILSLAND, ELIZABETH; SUNNERHAGEN, PER; NEVES, BRUNO JUNIOR; ANDRADE, CAROLINA HORTA; LEMOS CRAVO, PEDRO VITOR; MARANHAO COSTA, FABIO TRINDADE. Computational Chemogenomics Drug Repositioning Strategy Enables the Discovery of Epirubicin as a New Repurposed Hit for Plasmodium falciparum and P. vivax. Antimicrobial Agents and Chemotherapy, v. 64, n. 9 SEP 2020. Web of Science Citations: 1.

FERREIRA, LETICIA T.; VENANCIO, VINICIUS P.; KAWANO, TAILA; ABRAO, LAILAH C. C.; TAVELLA, TATYANA A.; ALMEIDA, LUDIMILA D.; PIRES, GABRIEL S.; BILSLAND, ELIZABETH; SUNNERHAGEN, PER; AZEVEDO, LUCIANA; TALCOTT, STEPHEN T.; MERTENS-TALCOTT, SUSANNE U.; COSTA, FABIO T. M. Chemical Genomic Profiling Unveils the in Vitro and in Vivo Antiplasmodial Mechanism of Acai (Euterpe oleracea Mart.) Polyphenols. ACS OMEGA, v. 4, n. 13, p. 15628-15635, SEP 24 2019. Web of Science Citations: 0.

BILSLAND, ELIZABETH; TAVELLA, TATYANA A.; KROGH, RENATA; STOKES, JAMIE E.; ROBERTS, ANNABELLE; AJIOKA, JAMES; SPRING, DAVID R.; ANDRICOPULO, ADRIANO D.; COSTA, FABIO T. M.; OLIVER, STEPHEN G. Antiplasmodial and trypanocidal activity of violacein and deoxyviolacein produced from synthetic operons. BMC Biotechnology, v. 18, APR 11 2018. Web of Science Citations: 2.

BILSLAND, ELIZABETH; VAN VLIET, LIISA; WILLIAMS, KEVIN; FELTHAM, JACK; CARRASCO, MARTA P.; FOTORAN, WESLEY L.; CUBILLOS, ELIANA F. G.; WUNDERLICH, GERHARD; GROTLI, MORTEN; HOLLFELDER, FLORIAN; JACKSON, VICTORIA; KING, ROSS D.; OLIVER, STEPHEN G. Plasmodium dihydrofolate reductase is a second enzyme target for the antimalarial action of triclosan. SCIENTIFIC REPORTS, v. 8, JAN 18 2018. Web of Science Citations: 6.