Cell Death and Inflammatory Signalling
Mitochondrial apoptosis and inflammasome activation Suitability: Honours, PhD/Doctorate Project leader: Dr Kate Lawlor e: kate.lawlor@hudson.org.au
Defining regulators of cell death and inflammasome activation Suitability: PhD/Doctorate, Honours Project leader: Dr Kate Lawlor e: kate.lawlor@hudson.org.au Project description: Pattern recognition receptors, including Toll-like receptors (TLRs) and NOD-like receptors (NLRs) are key components of the innate immune response. They sense microbial, host derived and environmental danger molecules, and induce inflammatory signalling responses, via inflammasomes and other molecular complexes. We recently defined how deficiency in the cell death inhibitory protein XIAP sensitises innate immune cells to TLR-induced NLRP3 inflammasome activation (Lawlor KE et al. Nature Comms 2015, Lawlor KE* et al. Cell Reports 2017). The aim of this project is to further define molecules, like XIAP, that regulate this alternative inflammasome pathway. This project offers the opportunity to be trained in a variety of techniques, including cell culture, Western blotting/immunoprecipitation, proteomics, overexpression/CRISPR Cas9 gene editing, flow cytometry, ELISA and qPCR. Keywords: Cell death, Inflammasomes, Innate immunity, infection, type I IFN, signal transduction
Identifying mitochondrial factors that activate inflammatory signalling Suitability: PhD/Doctorate, Honours Project leader: Dr Kate Lawlor e: kate.lawlor@hudson.org.au Project description: Mitochondrial (“intrinsic” BCL-2 family regulated) apoptosis has long been thought to be immunologically silent. However, using small molecule inhibitors of pro-survival BCL-2 family members, we have recently discovered that mitochondrial apoptosis can induce a cascade of events that culminate in activation of the NOD-like receptor 3 (NLRP3) inflammasome and proinflammatory cytokine, IL-1beta (Cell Reports 2018). In this project we will further characterise this pathway and test whether its activation alters cancer progression in vivo. This project will use our novel gene knockout macrophages and specific targeted drugs, plus a range of cell biology, biochemical and molecular approaches (e.g. inflammasome/cell death assays, ELISA, Western blotting, CRISPR Cas9 gene editing screens, proteomics). Keywords: Cell death, Cell signalling pathways, Inflammasomes, Innate immunity, Cancer
Project description: Macrophages are innate immune cells that detect environmental, pathogen or host cellular danger molecules, and initiate appropriate immune responses. We have recently discovered that targeting pro-survival proteins BCL-XL and MCL-1 in macrophages induces apoptosis to clear microbial infection (Speir M et al. Nature Microbiology 2016) and also triggers inflammation via activation of the NOD-like receptor 3 (NLRP3) inflammasome and Interleukin-1beta (Cell Reports 2018). This project aims to define novel regulators of this pathway and investigate how these proteins alter pathogen clearance. This project will use our novel gene knockout macrophages and specific targeted drugs, as well as a range of cell biology and biochemical/molecular approaches (e.g. inflammasome/cell death assays, ELISA, Western blotting, Q-PDR, over-expression systems, CRISPR Cas9 gene editing, infectious preclinical models). Keywords: Cell death, Cell signalling pathways, Inflammasomes, Innate immunity, Infectious Diseases
Role of pore forming proteins in Type 2 Diabetes Suitability: PhD/Doctorate Project leaders: Dr Kate Lawlor, Dr Hazel Tye e: kate.lawlor@hudson.org.au Project description: Dietary danger molecules, such as the saturated fatty acid palmitate, trigger the activation of the pro-inflammatory cytokine, interleukin-1β (IL-1β), via NLRP3 inflammasome activation. Based on our past work (Lawlor KE Nature Commun 2015 6:282) we recently discovered that extrinsic apoptotic caspase-8 activity in myeloid cells may drive this pathogenic NLRP3 inflammasome activation in a model of Type 2 Diabetes. Gasdermins (GSDMs) are a family of membrane pore-forming proteins that have recently been defined as proteolytic caspase substrates that induce a lytic form of cell death called pyroptosis. Specifically, inflammasome-associated caspase-1 and apoptotic caspase-8 have been shown to cleave GSDMD (Kayagaki N Nature 2015 526:666; Orning P Science 2018 362:1064), and apoptotic effector caspase-3 cleaves GSDME (Wang Y Nature 2017 547:99) to allow the release of inflammatory damage associated molecular patterns (DAMPS). The aim of this project is to define, using our GSDMD-, GSDME- and GSDMD/E-deficient mice, whether GSDM-dependent pyroptotic cell death contributes to NLRP3 inflammasome activation and/or DAMP release to worsen obesity-induced Type 2 Diabetes. This project offers the opportunity to be trained in a variety of techniques, including cell culture, Western blotting, inflammasome/cell death assays, ELISA, qPCR, tissue analysis-histology, flow cytometry, serum/liver/adipose metabolic assays, Type 2 diabetes preclinical models. Keywords: Type 2 Diabetes, Inflammation, IL-1, inflammasomes, immunometabolism, Gasdermins 7
Centre for Innate Immunity and Infectious Diseases | Student Research Projects 2023
