Award Orals

glomerulonephritis. LynKO.Monk-var mice had attenuated BCR calcium fluxing compared with LynKO mice, associated with restored expression of the surface IgM/CD21.35 coreceptor. Homeostatic LynKO.Monk-var mice had increased BCR lambda light chains reflecting increased endogenous receptor editing and thus B cell tolerisation. However, autoantibody levels were only modestly reduced compared with LynKO mice. Whilst BCR responses were attenuated, Monk-var mice had increased TLR responses. Monk directly modifies MyD88-dependent TLR signalling. We hypothesise that this role in MyD88 signalling forms the basis for Monk-var completely preventing development of glomerulonephritis in LynKO mice. Conclusions: We show that Monk-var attenuates autoimmunity in SLE-prone LynKO mice and prevents glomerulonephritis through a novel mechanism. We identify a novel role for Monk in regulating MyD88 signalling and identify Monk as a therapeutic target for treatment of LN and is the subject of preliminary drug development. Aim: Gasdermin-D (GSDMD) and pyroptosis can be to treated acute kidney Background: Pyroptosis, a pro-inflammatory form of cell death and is dependent on pore formation by cleaved Gasdermin-D. We hypothesis pyroptosis is important in ischemia-reperfusion cells (Foxp3-CD45RA + ) and PopV(CD45RA + Foxp3) as naïve effector CD4 + T cells. Methods: Peripheral blood mononuclear cells (PBMC) isolated from healthy Volunteers (HV, n = 11) and patients with kidney transplant surviving>10 yrs (RT, n = 15) were stained with panels of monoclonal antibodies to CD3, CD4, CD25, CD127, CD45RA, Foxp3, and CD19. Data was acquired on FACS CantoII using FACS DIVA(V8.0) and analysed using FlowJo. Lymphocytes were analysed after FSC vs SSC gating and doublets exclusion. Results: Lymphocyte and CD4 + T cells in HV and RT were similar, but RT had fewer B cells ( p < 0.05). Treg in RT was less than in HV ( p < 0.05). Naïve Treg (Pop I) ( p < 0.01) and naïve effector CD4 + T cells (Pop V) were lower in RT than HV. However, activated Treg (Pop II and III) were greater in RT ( p < 0.05), as was activated effector T cells(Pop IV) RT ( p < 0.05). Ratio of activated Treg (Pop II + III) to B cells was markedly greater in RT ( p < 0.05). Conclusions: Long surviving transplant patients had reduced naïve Treg and naïve effector cells but increased activated Treg and effector cells. The marked increase in the ratio of activated effector Treg to B cells increased suppression. Further studies are needed to establish the relevance of such changes to induction of tolerance. Background/aims: Maternal obesity adversely impacts metabolic health in mothers and offspring. Maternal complications include gestational diabetes and preeclampsia. Offspring have increased Background: The Cox proportional hazards model is commonly used to compare survival between the waiting list (WL) and after deceased donor kidney transplantation (TP). However, the Cox model only allows estimation of relative but not absolute survival. Flexible parametric models (FPM) solve this by modelling the baseline hazard which enables the creation of an absolute survival prediction for individuals with different characteristics. Aims: To develop FPMs for predicting survival on the waiting list versus deceased donor kidney transplantation. Methods: Using the Australia and New Zealand Dialysis and Transplant (ANZDATA) Registry, we included Australian adults waitlisted for first kidney-only deceased donor transplants over 2007 – 2020. We developed FPMs for waitlist and post-transplant survival. Covariates were decided using backwards elimination and the baseline hazard function was modelled using cubic splines. Results: 7552 patients were included in this analysis: 5429 (72%) received a deceased donor kidney transplant. The models were adjusted for age, gender, primary kidney disease, dialysis duration, comorbidities, and smoking status. The FPM allowed calculations of individual mean life expectancy. Aim: To develop a high-throughput platform to map the functional consequence and pathogenicity of genetic variants that contribute to CKD in Indigenous populations. Background: Indigenous Australian populations have the highest recorded rates of CKD worldwide with genetics identified as a major contributor. We have established a national collaboration to sequence and define the genetic contribution to the high rates of CKD in Indigenous communities. Methods: We designed a high-throughput screening platform utilizing cell lines, each capable of assaying four biochemically relevant immu-nological pathways simultaneously (eight total), utilizing distinct luciferase gene emission spectra. The lines were generated by stably transfecting human embryonic kidney (HEK293) cells with a Multiplex Luciferase Reporter (MLR) plasmid. Luciferase activity correlates with gene expression driven by promoters responsive to major disease-relevant pathways (e.g. NFkB, TGF β , IFN β , NFAT; or IFN α , STAT1-GAS, STAT3-site, AP1). Reporter lines expressing wild-type or variant genes are treated with a range of stimuli and biochemical pathway signalling differences determined from luciferase readouts. Results: We identified rare, potentially deleterious variants in CKD-relevant genes at high frequencies in patients from the Tiwi Islands including A20/TNFAIP3, AIRE, BLK, IRF5 as well variants in IRAK3, LYN, TRAF5 and RELB which are unique to the Tiwi. Variants in A20, LYN and AIRE were validated for altered function. Whilst the LYN variant had no impact, one variant in AIRE and two in A20 impaired IFN β and NFkB expression respectively, highlighting the feasibility of multiplexing to rapidly screen for deleterious variants in inflammatory cell cell communications sEV in the wave of patient-derived cultured and (1% O ₂ on Transwell inserts to separate sEV secreted analysed (qNano) and (miRNA sequencing). We profiled the injury in autologous PTEC co-cultured with harvested sEV using assays of DNA damage/cell death (flow cytometry, Western blot). Results: Significantly increased numbers of sEV were secreted from the apical surface of hypoxic PTEC compared with normoxic PTEC. No significant differences in basolateral sEV numbers were observed between culture conditions. Biological pathway analysis of apical hypoxic sEV cargo found distinct miRNA signatures associated with processes of cellular injury/disease. Apical sEV pro-duced by hypoxic PTEC selectively induced DNA damage ( γ -H2AX), cellular necrosis ( " Annexin-V + /propidium iodide + cells) and ferroptotic cell death ( # glutathione peroxidase-4, " 4-hydroxy-nonenal) in autologous PTEC compared with apical normoxic sEV and basolateral sEV. Conclusions: Our data establish PTEC-derived apical sEV as a key trigger of tubular death/loss in hypoxic CKD. This innovative con-cept of how tubular injury is propagated from the initiating insult into a ‘ wave of death ’ will have significant impact for CKD therapeutics. pathogenic mechanisms. combined inhibition. Furthermore, early both the of Background: Reduced oxygen levels during renal ischemia can induce AKI and lead to CKD through maladaptive repair. Tubulointerstitial fibrosis is a chronic and progressive feature of CKD, for which we lack effective treatments. Aim: To develop a human kidney organoid model of hypoxia-induced fibrosis to facilitate therapeutic studies. Methods: Human iPSC-derived organoids were generated from three different cell lines and cultured for 18d, then subjected to hypoxia (1%O2) or normoxia (21%O2) for 48 h (d20) followed by 5d recovery period in normoxia (d25). Organoids were examined by gene expression profiling, immunohistochemistry and immunofluorescence microscopy. Results: Robust hypoxic response was evident on d20 in mRNA levels of hypoxia-inducible genes (>5-fold increase in VEGFA, HK1 p < 0.001). Tubular damage marker KIM1/HAVCR1, stress-response gene JUN, and profibrotic signal TGFB1 were all upregulated (>2 fold p < 0.001) in hypoxic organoids on d20, consistent with an AKI response. Mitochondrial dysfunction was evident in hypoxic organoids on d20 with a 50% decrease ( p < 0.001) of PPARGC1A levels. On d25, expression of hypoxia-responsive genes and AKI markers returned to control levels. we show kidney organoids transiently upregu-late markers of AKI in response to hypoxia and later display signatures of maladaptive repair including elevated levels of ECM. These results suggest kidney organoids may aid in disease modelling and drug development for AKI and fibrosis. performed on the kidney cortex using RNA-sequencing. Results: We found senescence marker, p21, to be upregulated in the diabetic kidney, with a concomitant increase in senescence-associated secretory phenotype marker, IL-6. These markers were attenuated by the inhibition of C5aR1. Importantly, the levels of p21 correlated significantly with albuminuria in these mice. RNA-sequencing revealed pathways associated with TP53-regulated transcription of cell cycle genes was upregulated in diabetes and downregulated by PMX53. Conclusions: TP53 encodes for p53 which induces cell cycle arrest via p21. Collectively, these results suggest a role for C5aR1 in mediating cellular senescence in diabetic kidney disease. therapy of We conducted a clinical audit on patients with a recorded diagnosis of ADPKD in Southern Tasmania between 2013 and 2022. loop diuretic use (adjOR 0.38; 95%CI 0.15 – 0.99; p < 0.05) reduced the likelihood of requiring RRT. Comorbid IHD ( p = 0.03), diabetes mellitus ( p = 0.02), cerebrovascular accident ( p = 0.03), ventricular assist device use ( p = 0.03) and increasing CPB time ( p < 0.001) were all associated with remaining on RRT long-term. key associated and post-cardiac mofetil, cyclophosphamide, tacrolimus and rituximab. Belimumab therapy improved proteinuria, renal function, serological markers of SLE and disease activity scores in the two male patients, permitting a reduction in prednisolone dosing. One 124