Tackling Cancer with Yeast-Based Technologies Raphael Ferreira, Angelo Limeta, Jens Nielsen  Trends in Biotechnology  Volume 37, Issue 6, Pages 592-603 (June 2019) DOI: 10.1016/j.tibtech.2018.11.013 Copyright © 2018 Elsevier Ltd Terms and Conditions

Figure 1 Four Axes of Yeast’s Contribution to Cancer Research. (A) Elucidation of the mechanism of action of tumorigenesis using yeast relying on evolutionary conservation of pathways. Illustrated by loss of Beclin-1 deficiency, which in humans triggers tumorigenesis in certain cell types. Beclin-1 was found to be homologous to the yeast gene ATG6, whose loss prevents the correct extension of the autophagosome, leading to loss of autophagy. This finding ultimately linked autophagy to the tumor-forming properties of Beclin-1 [69]. BECN1, Beclin-1. (B) Chemical genetics-based small-molecule drug assay for target identification. The assay utilizes deletion or overexpression yeast libraries to retrieve targets that are sensitive or resistant to the drug treatment. (C) Metabolic engineering strategy for anticancer drug production. Insertion of heterologous genes from different organisms to allow complete rewiring of yeast metabolism [28]. (D) Yeast-based vaccination through the overexpression of tumor antigens displayed at the cell membrane. Heat-killed engineered yeast is recognized by immune cells, which triggers MHC I and II. MHC activation triggers T cell specialization against the antigen also displayed by the malignant cells [45]. Trends in Biotechnology 2019 37, 592-603DOI: (10.1016/j.tibtech.2018.11.013) Copyright © 2018 Elsevier Ltd Terms and Conditions

Figure 2 Yeast-Based Cancer Studies. Top: Synthetic lethality interaction screening between tumor suppressive genes (TSGs) and druggable target genes (DTs) for precision cancer therapy. (A) Synthetic genetic array (SGA) screening for DT and TSG interactions across four conditions, yielding a total of ∼169000 interactions, was first performed in yeast. MATa dtΔ, haploid yeast strains with orthologs of DTs deleted; MATα tsgΔ, haploid yeast strains with orthologs of TSGs deleted. (B) The top interactions that held true across conditions were kept for a drug TSG–knockdown interaction screen in HeLa cells (∼2000 interactions). IC20 and IC40, concentrations required for 20% and 40% inhibition, respectively. (C) Interaction network for one cross-species synthetic lethal interaction between the TSG XRCC3 and the histone deacetylase inhibitors vorinostat and rocilinostat. These novel interactions were validated through clonogenic assay. Square nodes represent DT genes. Saccharomyces cerevisiae gene names are below human gene names in italics. Bottom: Humanized antibody (trastumazab) production in glycoengineered Pichia pastoris. Glycoengineering of the trastuzumab antibody allows humanization of the glycosylation patterns of P. pastoris through: (i) deglycosylation of mannose repeats with endoH; and (ii) transglycosylation of desired glycans by endoS2. SHM-trastuzumab, trastuzumab with super-high-mannose glycans; SCT-trastuzumab, trastuzumab with sialylated biantennary complex glycans; blue square, N-acetylglucosamine; green circle: mannose; yellow circle, galactose; purple diamond, sialic acid. Trends in Biotechnology 2019 37, 592-603DOI: (10.1016/j.tibtech.2018.11.013) Copyright © 2018 Elsevier Ltd Terms and Conditions

Figure 3 Yeast-Based Milestones That Had a Direct or Indirect Impact on Cancer Research. Nonexhaustive timeline of accomplishments, events, and/or publications in fundamental biology as well as cancer biology using Saccharomyces cerevisiae, Schizosaccharomyces pombe, and Pichia pastoris. Arrow with square end point, milestone with an indirect impact on cancer research; arrow with diamond end point, milestone with a direct application in cancer. Trends in Biotechnology 2019 37, 592-603DOI: (10.1016/j.tibtech.2018.11.013) Copyright © 2018 Elsevier Ltd Terms and Conditions

Figure I Examples of Future Yeast-Based Applications. Top: Point-of-care testing through yeast-based G protein-coupled receptor (GPCR) biosensor detection of volatile organic compounds (VOCs) released by tumor cells. Bottom: Detection and eradication of tumor cells using a T cell receptor (TCR) and cytotoxin yeast-based system. Trends in Biotechnology 2019 37, 592-603DOI: (10.1016/j.tibtech.2018.11.013) Copyright © 2018 Elsevier Ltd Terms and Conditions