Reference number : GC-2017-Q1-01774, amendment number : , agreement type : G, recipient legal name : Yates, Robin (University of Calgary), recipient country : CA, recipient province : nan, recipient city : Alberta, recipient postal code : nan, program name english : Discovery Grants Program - Individual, program name french : Programme de subventions à la découverte - individuelles, program purpose english : Phagosomes are organelles that are formed after the engulfment of material by phagocytes such as macrophages and dendritic cells. The lumen of phagosome is a dynamic and complex microenvironment. It is the site of microbial killing, precise proteolytic processing of antigens and the degradation and recycling of macromolecules during tissue remodeling, growth and homeostasis. Despite the importance of the phagosome, there are critical gaps in our understanding of its function and dysfunction. One particular gap is the impact of redox chemistries on phagosomal function. 
 Over the past seven years, my group has investigated the redox control of critical chemistries within phagosomes and lysosomes. We discovered that NOX2 oxidizes the normally reductive lumen of the phagosome and inhibits both oxidation-sensitive proteolysis and disulfide reduction&mdash;two key chemistries for antigen processing and protein turn-over. However, the other side to this story was yet to be told: what re-reduces these oxidized proteases and how is the reductive capacity of phagosomes and lysosomes maintained? We recently addressed this by conducting a screen that identified a cytosolic pathway that utilizes NADPH and a selenoprotein reductase to maintain the reductive environment within the phagosome (30). This study was highlighted as &ldquo;Leading Edge Research&rdquo; in the July 2016 issue of the Journal of Leukocyte Biology with a dedicated editorial. The authors of the editorial stated: that &ldquo;[while the study] represents an excellent starting platform to discern how the reductive capacity of the phagosome is maintained&rdquo;&mdash;&ldquo;[it] is not the final conclusion to the story&rdquo; (29). We agree. 
 This proposal aims to continue the interrogation of the fundamental biochemical processes that promote essential reductive chemistries in phagosomes. This is critically needed in the field of phagocyte biology, but will also likely uncover key pathways that are necessary to maintain the redox balance in endosomes and lysosomes&mdash;common to all eukaryotic cells. Additionally, my research program will be broadened beyond the phagosome, as we propose to adapt our technologies to explore the control of reductive chemistries in a new cellular location&mdash;the osteoclast lacunae. The three aims propose a breadth of unique and cutting edge technologies to test specific hypotheses and facilitate discovery-based knowledge creation. 
 
 Aim 1: Specific identification of components of the newly identified cytosolic pathway that provides reductive equivalents to the phagosomal lumen. 
 Aim 2: Identification of upstream and downstream interacting partners of GILT. 
 Aim 3: Exploration of the role of GILT and reductive pathways in the lacunae of the osteoclast. 
 
 Funding of this proposal will allow my group to build upon our strong track record of pioneering new research areas in phagocyte biology and creating an engaging, rigorous and unique training environment., program purpose french : Phagosomes are organelles that are formed after the engulfment of material by phagocytes such as macrophages and dendritic cells. The lumen of phagosome is a dynamic and complex microenvironment. It is the site of microbial killing, precise proteolytic processing of antigens and the degradation and recycling of macromolecules during tissue remodeling, growth and homeostasis. Despite the importance of the phagosome, there are critical gaps in our understanding of its function and dysfunction. One particular gap is the impact of redox chemistries on phagosomal function. 
 Over the past seven years, my group has investigated the redox control of critical chemistries within phagosomes and lysosomes. We discovered that NOX2 oxidizes the normally reductive lumen of the phagosome and inhibits both oxidation-sensitive proteolysis and disulfide reduction&mdash;two key chemistries for antigen processing and protein turn-over. However, the other side to this story was yet to be told: what re-reduces these oxidized proteases and how is the reductive capacity of phagosomes and lysosomes maintained? We recently addressed this by conducting a screen that identified a cytosolic pathway that utilizes NADPH and a selenoprotein reductase to maintain the reductive environment within the phagosome (30). This study was highlighted as &ldquo;Leading Edge Research&rdquo; in the July 2016 issue of the Journal of Leukocyte Biology with a dedicated editorial. The authors of the editorial stated: that &ldquo;[while the study] represents an excellent starting platform to discern how the reductive capacity of the phagosome is maintained&rdquo;&mdash;&ldquo;[it] is not the final conclusion to the story&rdquo; (29). We agree. 
 This proposal aims to continue the interrogation of the fundamental biochemical processes that promote essential reductive chemistries in phagosomes. This is critically needed in the field of phagocyte biology, but will also likely uncover key pathways that are necessary to maintain the redox balance in endosomes and lysosomes&mdash;common to all eukaryotic cells. Additionally, my research program will be broadened beyond the phagosome, as we propose to adapt our technologies to explore the control of reductive chemistries in a new cellular location&mdash;the osteoclast lacunae. The three aims propose a breadth of unique and cutting edge technologies to test specific hypotheses and facilitate discovery-based knowledge creation. 
 
 Aim 1: Specific identification of components of the newly identified cytosolic pathway that provides reductive equivalents to the phagosomal lumen. 
 Aim 2: Identification of upstream and downstream interacting partners of GILT. 
 Aim 3: Exploration of the role of GILT and reductive pathways in the lacunae of the osteoclast. 
 
 Funding of this proposal will allow my group to build upon our strong track record of pioneering new research areas in phagocyte biology and creating an engaging, rigorous and unique training environment., agreement title english : Deciphering the reductive pathways in phagosomes and lysosomes., agreement title french : Deciphering the reductive pathways in phagosomes and lysosomes., agreement number : RGPIN, agreement value : 200000.0, agreement start date : 2017-05-10, agreement end date : nan, description english : nan, description french : nan, expected results english : nan, expected results french : nan, additional information english : Grant or Award spanning more than one fiscal year. (2017-2018 to 2022-2023), additional information french : Subvention ou bourse octroyée s'appliquant à plus d'un exercice financier. (2017-2018 à 2022-2023), owner organization : nserc-crsng, owner organization title : Natural Sciences and Engineering Research Council of Canada | Conseil de recherches en sciences naturelles et en génie du Canada