uPAR Research Articles

Self-rated health and chronic inflammation are related and independently associated with hospitalization and long-term mortality in the general population

The subjective indicator of health self-rated health (SRH) and the chronic inflammation biomarker soluble urokinase plasminogen activator receptor (suPAR) are both robust predictors of healthcare use and mortality. However, the possible relationship between SRH and suPAR in the assessment of hospitalization and mortality risk is unknown. We used data from the Danish population-based Inter99 cohort to examine the association between SRH and suPAR and test their individual and combined associations with 2-year risk of acute hospitalization and 5- and 15-year mortality. SRH and serum suPAR levels were measured in 5490 participants (median age 45.1 years, 48.7% men). Poorer SRH was associated with elevated suPAR. In unadjusted analyses, SRH and suPAR were individually associated with higher risks of acute hospitalization and mortality, and both measures remained independently associated with higher risks of hospitalization and 15-year mortality after mutual adjustments. The association of suPAR with mortality was stronger in poorer SRH categories, and when combined, SRH and suPAR could identify different groups of individuals with increased risk of acute hospitalization and mortality. Both SRH and suPAR were independently associated with risk of acute hospitalization and mortality, and different combinations of the two measures could identify different groups of individuals at increased risk.

Identifying Mechanisms of Resistance to DHODH Inhibition in Cancer

All transcriptionally active and rapidly dividing cells require high amounts of pyrimidines for DNA replication and repair and RNA synthesis. The only two known mechanisms by which cells can acquire pyrimidines are the de novo pyrimidine biosynthesis pathway and the pyrimidine salvage pathway. The rate-limiting enzyme in de novo synthesis is dihydroorotate dehydrogenase (DHODH), a critical metabolic enzyme that converts dihydroorotate (DHO) to orotate, which is then converted to the pyrimidine precursor uridine monophosphate (UMP). The rate-limiting enzyme in the salvage pathway is uridine-cytidine kinase 2 (UCK2), a ribonucleoside kinase that converts uridine to UMP and cytidine to cytidine monophosphate (CMP). Under physiologic conditions, salvage pathway substrates are limiting, and many cancer types, most notably acute myeloid leukemia (AML), are highly dependent on DHODH activity for proliferation and survival. Based on these observations, several DHODH inhibitors have been developed as anti-cancer therapeutics and have been tested clinically. Unfortunately, to date, these drugs have failed to show significant anti-cancer activity in early-phase clinical trials. In order to understand how leukemia cells that are highly sensitive to DHODH inhibitors can bypass their dependence on DHODH activity, we conducted positive-selection whole-genome CRISPR-activation drug resistance screens in TF-1 human AML cells using BAY 2402234, a highly potent and highly specific DHODH inhibitor. In preliminary studies, we found that 1 nM BAY 2402234 fully inhibits TF-1 cells whereas TF-1 cells that overexpress high levels of DHODH require 50 nM BAY 2402234 to achieve the same effect. Importantly, the effects of 1 nM and 50 nM BAY 2402234 are fully rescued by addition of uridine to the culture media, demonstrating that the anti-leukemic activity of BAY 2402234 is on-target at both drug doses. To perform the drug resistance screens, we stably expressed nuclease-deactivated Cas9 fused to a transcriptional activator (dCas9-VP64) in TF-1 cells and then infected the cells with the human whole-genome Calabrese sgRNA lentiviral library. We then passaged the cells in media containing vehicle control (DMSO), 1 nM or 50 nM BAY 2402234 for 18 days, collecting cells on day 0 and day 18 of drug treatment. Genomic DNA was then extracted and was subjected to next generation sequencing to assess the relative abundance of each sgRNA in each sample. We reasoned that activation of genes that enhance de novo pathway activity at the level of, or downstream of, DHODH would promote resistance to 1 nM BAY 2402234 but not 50 nM BAY 2402234, whereas activation of genes that act through de novo pathway-independent mechanisms would promote resistance to both 1 nM and 50 nM BAY 2402234. As predicted, sgRNAs targeting DHODH conferred resistance to 1 nM but not 50 nM BAY 2402234 whereas sgRNAs targeting UCK2 conferred resistance to both 1 and 50 nM BAY 2402234 (Figure 1). A number of other known salvage pathway genes scored in both screens, including uracil phosphoribosyltransferase (UPRT) and the nucleoside transporters SLC4A1 and SLC29A1. Unexpectedly, the screens also identified carbamoyl-phosphate synthetase 2/aspartate transcarbamoylase/dihydroorotase (CAD) as a potential mechanism of resistance to DHODH inhibition. CAD is a trifunctional multi-domain enzyme that acts upstream of DHODH, catalyzing the first three steps in de novopyrimidine biosynthesis. How upregulation of CAD activity can bypass the requirement for DHODH activity is not clear, although CAD is a very large enzyme complex about which very little is known besides its de novo pyrimidine synthesis activity. The screens also identified several genes that are not known to play a role in pyrimidine metabolism as potential resistance mechanisms. These include the serine-threonine kinase MARK3, the serine-threonine phosphatase CTDSPL2, and the RNA-binding protein NANOS2. Finally, the screens identified several genes whose upregulation sensitized cells to BAY 2402234, including the urokinase receptor PLAUR, the adaptor protein BCAR3, the scaffold protein SHOC2, and the deoxynucleoside phosphohydrolase SAMHD1. Studies are currently underway to validate the results of the CRISPRa screens and elucidate the mechanisms by which the candidate resistance genes help cells to bypass their requirement for DHODH activity. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Abstract 10108: SuPAR, Galectin-3 and ST2 Are Associated With Chronic Kidney Disease Progression in Patients With Heart Failure With Reduced Ejection Fraction

Introduction: Patients with heart failure with reduced ejection fraction (HFrEF) are at risk for both acute and progressive kidney disease. Elevated levels of the circulating soluble urokinase plasminogen activator receptor (suPAR), Galectin-3, and soluble suppression of tumorigenicity 2 (ST2) have been associated with kidney disease progression in those with CKD. However, there is little known about their predictive value for glomerular filtration rate (GFR) decline in populations with HFrEF. Methods: We aimed to determine whether these biomarkers were associated with decline in GFR and death in patients with HFrEF. We included 310 participants from the Registry Evaluation of Vital Information for Ventricular Assist Devices in Ambulatory Life in whom baseline suPAR, Galectin-3, and ST2 and repeated measures of GFR were obtained. The primary outcome was change in 2021 CKD-EPI creatinine-based GFR over 2 years. Secondary outcome was mortality. A joint cox regression model for mortality controlled for age, sex, race, NYHA class, and change in GFR and accounted for competing risks of ventricular assist device (VAD) implantation and heart transplantation. Results: Participants were followed for 2 years or until VAD, transplant, or death. Mean age was 59 years (standard error of mean 0.7) and 36% had diabetes mellitus. Median GFR was 60 ml/min/1.73m 2 with 58% NYHA Class III. Of these, 45 died, 33 received LVAD, and 25 were transplanted. Higher baseline suPAR (β coefficient -0.22 √(ml/min/1.73m 2 ); 95%CI -0.29, -0.15; P<0.001), Galectin-3 (β coefficient -0.02 √(ml/min/1.73m 2 ); 95%CI -0.04, -0.01; P=0.012), and ST2 (β coefficient -0.01 √(ml/min/1.73m 2 ); 95%CI -0.02, -0.01, -0.19; P<0.001) plasma concentrations were associated with a decline in eGFR over time in multivariable models. Only ST2 (HR 1.02 per ng/mL increase; 95%CI 1.01, 1.03; P<0.001) was associated with mortality, after controlling for longitudinal change in GFR. Conclusions: Higher baseline suPAR, Galectin-3, and ST2 were associated with GFR decline over 2 years in patients with HFrEF. ST2 was associated with a greater risk of mortality, while suPAR and Galectin-3 were not. These biomarkers may be incremental tools used to predict GFR decline in patients with advanced heart failure.

Ability of combined soluble urokinase plasminogen activator receptor to predict preventable emergency attendance in older patients in Japan: a prospective pilot study.

Soluble urokinase plasminogen activator receptor (suPAR) is a strong and nonspecific inflammatory biomarker that reflects various immunologic reactions, organ damage, and risk of mortality in the general population. Although prior research in acute medical patients showed that an elevation in suPAR is related to intensive care unit admission and risk of readmission and mortality, no studies have focused on the predictive value of suPAR for preventable emergency attendance (PEA). This study aims to evaluate the predictive value of suPAR, which consists of a combination of white blood cell count (WBC), C-reactive protein (CRP), and the National Early Warning Score (NEWS), for PEA in older patients (>65 years) without trauma who presented to the emergency department (ED). This single-center prospective pilot study was conducted in the ED of the Association of EISEIKAI Medical and Healthcare Corporation Minamitama Hospital, in Hachiouji City, Tokyo, Japan, from September 16, 2020, to June 21, 2022. The study included all patients without trauma aged 65 years or older who were living in their home or a facility and presented to the ED when medical professionals decided an emergency consultation was required. Discrimination was assessed by plotting the receiver-operating characteristic (ROC) curve and calculating the area under the ROC curve (AUC). During the study period, 49 eligible older patients were included, and thirteen (26.5%) PEA cases were detected. The median suPAR was significantly lower in the PEA group than in the non-PEA group (p<0.05). For suPAR, the AUC for the prediction of PEA was 0.678 (95% CI 0.499-0.842, p<0.05), and there was no significant difference from other variables as follows: 0.801 (95% CI 0.673-0.906, p<0.001) for WBC, 0.833 (95% CI 0.717-0.934, p<0.001) for CRP, and 0.693 (95% CI 0.495-0.862, p<0.05) for NEWS. Furthermore, the AUC for predicting PEA was 0.867 (95% CI 0.741-0.959, p<0.001) for suPAR + WBC + CRP + NEWS, which was significantly higher than that of the original suPAR (p<0.01). The cutoff values, sensitivity, specificity, and odds ratio of suPAR and suPAR + WBC + CRP + NEWS were 7.5 and 22.88, 80.6% and 83.3%, 53.8% and 76.9%, and 4.83 and 16.67, respectively. This study has several limitations. First, this was pilot study, and we included a small number of older patients. Second, the COVID-19 pandemic occurred during the study period, so that there may be selection bias in the study population. Third, our hospital is a secondary emergency medical institution, and as such, we did not treat very fatal cases, which could be another cause of selection bias. Our single-center study has demonstrated the moderate utility of the combined suPAR as a triage tool for predicting PEA in older patients without trauma receiving home medical care. Before introducing suPAR to the prehospital setting, evidence from multicenter studies is needed.

Serum Levels of Urokinase Plasminogen Activator Receptor (suPAR) Discriminate Moderate Uncontrolled from Severe Asthma

The most clinically useful concept in asthma is based on the intensity of treatment required to achieve good asthma control. Biomarkers to guide therapy are needed. To investigate the role of circulating levels of soluble urokinase plasminogen activator receptor suPAR as a marker for asthma severity. We recruited patients evaluated at the Asthma Clinic, University of Thessaly, Greece. Asthma severity and control were defined according to the GINA strategy and Asthma Contro Test (ACT). Anthropometrics, spirometry, fractional exhaled nitric oxide (FeNO), suPAR, blood cell count, c-reactive protein (CRP), and analyses of kidney and liver function were obtained. Patients with a history of inflammatory, infectious, or malignant disease or other lung disease, more than 5 pack years of smoking history, or corticosteroid therapy were excluded. We evaluated 74 asthma patients (69% female, mean age 57 ± 17 years, mean body mass index (BMI) 29 ± 6 kg/m2). In total, 24%, 13%, 6%, 5%, 29% and 23% of the participants had mild well-controlled, mild uncontrolled, moderate well-controlled, moderate uncontrolled, severe well-controlled, and severe uncontrolled asthma, respectively. Overall, 67% had T2-high asthma, 26% received biologics (15% and 85% received omalizumab and mepolizumab, respectively), and 34% had persistent airway obstruction. suPAR levels were significantly lower in asthmatics with moderate uncontrolled asthma than in patients with severe uncontrolled asthma without (2.1 ± 0.4 vs. 3.3 ± 0.7 ng/mL, p = 0.023) or with biologics (2.1 ± 0.4 vs. 3.6 ± 0.8 ng/mL, p = 0.029). No correlations were found between suPAR levels and age, BMI, T2 biomarkers, CRP, or spirometric parameters. suPAR levels were higher in asthmatics with severe disease than in those with moderate uncontrolled asthma.

ALG3 Promotes Peritoneal Metastasis of Ovarian Cancer through Increasing Interaction of α1,3-mannosylated uPAR and ADAM8.

Peritoneal metastasis is the main cause of poor prognoses and high mortality in ovarian cancer patients. Abnormal protein glycosylation modification is associated with cancer malignancy. Elevated α1,3-mannosyltransferase 3 (ALG3), which catalyzes the α1,3-mannosylation of glycoproteins, has been found in some malignant tumors. However, the pathological significance of ALG3 and its regulatory mechanism in ovarian cancer metastasis is unclear. The results showed that the level of ALG3/α1,3-mannosylation was higher in human ovarian cancer tissues compared with normal ovarian tissues, as measured by Lectin chip, Western blot and Lectin blot analyses, as well as ovarian tissue microarray analysis. ALG3 was also correlated with the poor prognosis of ovarian cancer patients, according to survival analysis. The downregulation of ALG3 decreased the proliferation, stemness and peritoneal metastasis of ovarian cancer cells. The increase in urokinase plasminogen activator receptor (uPAR) α1,3-mannosylation catalyzed by ALG3 enhanced urokinase plasminogen activator (uPA)/uPAR activation and the interaction of uPAR with a disintegrin and metalloproteinase 8 (ADAM8), which promoted ovarian cancer peritoneal metastasis via the ADAM8/Ras/ERK pathway. Furthermore, decreased ALG3 suppressed ascites formation and the peritoneal metastasis of ovarian cancer cells in mice. This study highlights ALG3 as a potential diagnostic biomarker and prospective therapeutic target for ovarian cancer.

Comparative performances of soluble urokinase plasminogen activator receptor and Mid-regional proADM to predict composite death and new heart failure rehospitalisation in acutely breathless patients

Abstract Background Soluble urokinase plasminogen activator receptor (suPAR) is a pleotropic receptor, capable of orchestrating plaque vulnerability and vascular immune dysfunction. Mid-regional pro-adrenomedullin (MR-proADM) is the stable peptide precursor of adrenomedullin, with concentrations reflective of vasodilation and cardiac remodelling. We compared the prognostic performances of suPAR and MR-proADM for the composite clinical endpoint of death and new heart failure (HF) in patients with undifferentiated breathlessness. Methods Patients presenting to hospital with the primary complaint of acute dyspnoea were recruited in New Zealand (n=612) and in Singapore (n=483)]. Baseline plasma measurements were undertaken for suPAR (ViroGates) and MR-proADM (Thermo Scientific). Cardiac biomarker levels of NT-proBNP (Roche) was available on all patients. Statistical assessment was made using SPSS v28 (IBM), with all biomarkers treated as continuous variables and presented as median [interquartile range (IQR)]. Prognostic performance of suPAR, MR-proADM and NT-proBNP to predict the composite clinical endpoint of death/new HF at 90-days and 1-yr were assessed using receiver operator curve (ROC) area under the curve (AUC) analysis (Z-scores) and Cox hazard regression analysis (per doubling of biomarker concentrations) after adjustment for traditional risk factors. P-value &amp;lt;0.05 was considered statistically significant. Results In the entire acutely breathless cohort [median age: 65 years (IQR: 52.9–76.0), 63.1% males], 343/1095 of patients had the final adjudication of ADHF. suPAR and MR-proADM concentrations were higher with increasing age (Spearmans rho, r&amp;gt;0.46, P&amp;lt;0.0001), lower eGFR (r&amp;gt;0.58, P&amp;lt;0.0001) and in those with ADHF (r&amp;gt;0.40, P&amp;lt;0.0001). During the follow-up period, 122 patients were categorised with death/new HF by 90-days, rising to 281 at 1-year. suPAR and MR-proADM were able to predict death/new HF at 90-days (both ROC-AUC &amp;gt;0.77) and at 1-year (both ROC-AUC ≥0.78) (Table 1). All markers were however less accurate in predicting this endpoint in the presence of ADHF (ROC-AUC &amp;lt;0.71). After adjustment in Cox-regression modelling, suPAR obtained HR &amp;gt;1.35 per doubling of suPAR concentrations (P=0.001) for outcomes at 90-days and at 1-year (Table 2), achieving the highest prognostic performance for this clinical endpoint, followed by NT-proBNP (HR &amp;gt;1.29) (Table 2), whilst MR-proADM was not an independent predictor of death/HF in this cohort. suPAR was also an independent predictor of death/HF for patients with ADHF, obtaining HR &amp;gt;1.35 per doubling of concentrations. Above a cut-off concentration of 3.6 ng/mL, suPAR was associated with a HR of 2.1 (95% CI: 1.55–2.91) for death/HF at 1-year for acutely dyspnoeic patients. Conclusion suPAR concentrations is superior than MR-proADM in predicting the clinical end-point of death/HF at 1-year in this cohort. It may aid in risk-stratification strategies for the management of acutely breathless patients. Funding Acknowledgement Type of funding sources: Foundation. Main funding source(s): National Heart Foundation of New ZealandHealth Research Council of New Zealand

Soluble urokinase plasminogen activator receptor predicts survival and hospitalization for heart failure in elective cardiac surgery

Abstract Aims The study investigated the prognostic value of soluble urokinase plasminogen activator receptor (suPAR) patients undergoing cardiac surgery and calculated a simplified biomarker score comprising suPAR, N-terminal pro B-type natriuretic peptide (NT-proBNP) and age. Methods and results Biomarkers were assessed in a cohort of 478 patients undergoing elective cardiac surgery. After a median follow-up of 4.2 years a total of 72 (15.1%) patients died. SuPAR, NT-proBNP and age were independent prognosticators of mortality in a multivariable Cox regression model after adjustment for EuroScoreII. We then calculated a simplified biomarker score comprising age, suPAR and NT-proBNP, which had a superior prognostic value compared to EuroScoreII (Harrel's C of 0.76 vs 0.72; P for difference = 0.02). Besides long-term mortality, the biomarker score had an excellent performance predicting one-year mortality and hospitalization due to heart failure. Conclusion The biomarker suPAR and NT-proBNP were strongly and independently associated with mortality in patients undergoing cardiac surgery. A simplified biomarker score comprising only 3 variables (age, suPAR and NT-proBNP) performed better than the established EuroScoreII with respect to intermediate and long-term outcome as well as hospitalization due to heart failure. As such, integration of established and upcoming biomarkers in clinical practice may provide improved decision support in cardiac surgery. Funding Acknowledgement Type of funding sources: None.

Determining biomarkers for evaluation and diagnosis of hereditary angioedema.

RationaleKallikrein‐bradykinin‐forming cascade is known to cause hereditary angioedema (HAE) acute angioedema (AE) attacks. Further research of HAE attacks is needed to explain disease heterogeneity, predict treatment response and identify biomarkers for monitoring HAE attacks. Differential expression of the microvascular endothelial cell‐surface receptors for example, g‐C1qR, cytokeratin‐1, and plasminogen‐activator‐urokinase‐receptor (PLAUR) were hypothesized as biomarkers of AE attacks.MethodTo understand HAE attacks, the differentially expressed genes (DEGs) in RNAseq and mi‐RNAseq data of total RNA extracted from skin biopsies of lesional versus non‐lesional skin collected during and between attacks in Type‐1 HAE patients (n = 11; F:M = 8:3) were compared. To understand the HAE variants, DEGs in skin biopsies from HAE with normal C1 inhibitor (n = 5, F:M = 5:0), and non‐HAE (n = 7; F:M = 3:4) patients were compared. Gene‐set enrichment analyses and regulator effects analysis of these DEGs identified biological pathways in HAE attacks and their regulators.ResultsPLAUR gene, encoding urokinase‐type plasminogen activator (u‐PAR), was constitutively over‐expressed in HAE‐Type‐1 versus non‐HAE controls suggestive of overactive u‐PAR‐mediated signaling via binding to Factor‐XII. Baseline PLAUR expression was associated with severe AE (p = 0.05). The 18 significant DEGs investigated between baseline and AE attack samples in Type1‐HAE were enriched in beta1/beta3‐integrin cell surface interactions and IL‐6‐mediated signaling. Regulator effects analysis suggests a role for IL‐1b in HAE flares. AKT2, the mRNA regulated by the differentially‐expressed miR‐184A, was also associated with HAE attacks.ConclusionAngiopoetin‐activated β1‐integrin signaling pathways causing endothelial destabilization, and avid binding of factor XII to u‐PAR are possible novel mechanisms for progression of the endothelial kinin‐bradykinin‐forming cascade in HAE attacks.

Evaluation of Serum Pentraxin-3 and suPAR Levels as Acute Phase Reactants in Patients with COVID-19

Coronavirus disease-2019 (COVID-19) is the most challenging health problem of our century, but our knowledge about the disease is limited. Most individuals infected with severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), which causes COVID-19, have mild symptoms such as headache, sore throat, joint pain, loss of sense of taste and smell. However, infection also causes significant morbidity and mortality, especially in individuals over 65 years of age with comorbidities. However, it is not known exactly which patients will have a poor prognosis. In this study, it was aimed to determine serum Pentraxin-3 (PTX3) and soluble urokinase plasminogen activator receptor (suPAR) levels in COVID-19 patients, and to evaluate the relationship between PTX3 and suPAR levels and the clinical status of the disease. This study was conducted with 150 patients who were confirmed to have COVID-19 by microbiological or clinical/radiological methods between April 1 and December 31, 2020. Thirty people with no known history or symptoms of COVID-19 and negative reverse transcription-polymerase chain reaction (RT-PCR) results also constituted the control group. Patients admitted to inpatient services due to COVID-19 constituted the service group (n= 75) and patients admitted to the intensive care unit (ICU) constituted the ICU group (n= 75). Serum PTX3 and suPAR levels were analyzed by enzyme-linked immunoassay (ELISA) and the results were compared between the three groups. The patients' leukocyte, neutrophil, neutrophil/lymphocyte ratio (NLR), troponin, procalcitonin (PCT), D-dimer, C-reactive protein (CRP), lymphocyte and ferritin results were included in the analysis. The mean age of the patients was 67.2 ± 11.8, and 62.0 ± 8.4 in the control group. There was no significant difference between the groups in terms of female/male ratio (p= 0.582). The PTX3 and suPAR levels of the patients were higher than the controls (p= 0.001, p= 0.023, respectively). PTX3 and suPAR levels were higher in the service group than the ICU group (p<0.001, p= 0.004, respectively) and the control group (p<0.001, p= 0.001, respectively). However, PTX3 (p= 0.291) and suPAR (p= 0.411) concentrations did not differ between ICU and control groups. The most determining parameters in ICU admission were found to be leukocytes (AUC= 0.840), neutrophils (AUC= 0.840), and NLR (AUC= 0.835), respectively. The most predictive parameters for mortality were PCT (AUC= 0.712), NLR (AUC= 0.708) and D-dimer (AUC= 0.695), respectively. In our study, serum PTX3 and suPAR concentrations were found to be high in COVID-19 patients. In patients admitted to the ICU, PTX3 and suPAR levels were observed at low levels. Low levels of PTX3 and suPAR in COVID-19 patients were thought to be clinically important.

Perspectives on Drug Development in Early ADPKD.

Perspectives on Drug Development in Early ADPKD.

WITHDRAWN: Soluble urokinase plasminogen activator receptor and its complicated role in hemodialysis (HD) patients with Covid-19 infection

Soluble urokinase plasminogen receptor (suPAR) is a protein in the blood that has been described to reflect the severity status of systemic inflammation. We investigated the association between admission suPAR levels and severity and outcome of HD patients with Covid-19 infection. In an observational study of adult HD patients hospitalized for Covid-19, we measured suPAR levels in plasma samples. The time table for those measurements were as follows: at the beginning of admission, after a hemoperfusion (HP) session for those patients that received them, and just before discharge. Of the 17 patients (7 were male), 13 patients received HP (mean age: 74 years old). The median suPAR level was 12.94ng/ml. For those who undertook HP in HD unit the median suPAR level before the session was 12.95ng/mil and 6.2ng/ml at the end of each session (p<0.05). 3 patients had a suPAR level below 7ng/ml. 2 of them survived without developing pleural effusions. 7 patients were discharged from the hospital with median suPAR level 12.08ng/ml which did not differ significantly from the median suPAR level of the deceased ones (13.68ng/ml). Admission levels of suPAR in HD patients hospitalized for Covid-19 do not seem to be predictive for their clinical course in general. Chronic Kidney Disease and its relation to suPAR independently of patients' inflammation status may be the key component for our notice. Despite that, in patients where low levels of suPAR combined with absence of pleural effusions the prognosis was excellent.

The Pattern of Soluble Urokinase Plasminogen Activator Receptor (suPAR) Levels in Lupus Nephritis and Its Correlation with Renal Biopsy

The Pattern of Soluble Urokinase Plasminogen Activator Receptor (suPAR) Levels in Lupus Nephritis and Its Correlation with Renal Biopsy

Piptide Chemotypes for Perturbation of the Interaction of Urokinase with Its Receptor

Only a few small molecules that disrupt the uPA and uPA receptor (uPAR) interaction have been discovered despite decades of research in the area, and none have been approved in clinical trials. Research reported here features two new ways of considering the problem of discovering small molecules to disrupt uPA•uPAR, specifically in terms of chemotype design and method of evaluation. Chemotypes used in this work are piptides (Arancillo . Angew. Chem., Int. Ed., 2021, 60, 6653-6659) with side chains corresponding to the uPA loop that binds uPAR. Further, hybrids of 1 and another uPAR ligand developed in these labs (2), i.e., 3 and 4, were also designed and tested. All the piptide chemotypes bound uPAR at concentrations of 50 μM or less. Members of this series had Ki values <3 μM and showed favorable responses in cellular assays; these data are comparable with the best small molecule uPA•uPAR disruptors in the literature (from conventional screening).

Interleukin-1β and plasminogen activating system members in endometriotic stromal cell migration/invasion

To study the role of interleukin (IL)-1β and the plasminogen activating (PA) system members in endometriotic stromal cell (ESC) migration/invasion. Primary cultures of ESCs. Tertiary referral center for endometriosis and pelvic pain. Patients with surgically excised endometriosis. Interleukin-1β stimulation of primary cultures of ESCs and knockdown of the PA system members urokinase plasminogen activator (uPA), uPA receptor, and plasminogen activator inhibitor-1 (PAI-1). Invasion/migration assays. In primary cultures, IL-1β-stimulated ESC production of the PA system members uPA, uPA receptor, and PAI-1. Interleukin-1β also enhanced ESC migration and invasion, and these effects were inhibited by the IL-1 receptor-1 antagonist anakinra. Knockdown of each of the 3 PA system members also inhibited ESC migration and invasion. Knockdown of these PA system members further attenuated the impact of IL-1β on migration and invasion, suggesting that they mediated the promigration and proinvasion effects of IL-1β. To supplement the cell culture work, immunohistochemistry was performed on tissue sections of endometriotic epithelium/stroma: uPA, PAI-1, and IL-1β histoscores were not found to be correlated with each other. In primary cultures of ESCs, IL-1β induces migration and invasion, which is mediated by PA system members and inhibited by the drug anakinra. However, the immunohistochemistry expression of IL-1β, urokinase plasminogen inhibitor-1, and PAI-1 were not correlated, suggesting other regulatory mechanisms for PA system members. Inhibition of IL-1β (e.g., with anakinra) may have potential as a novel treatment approach for the migration/invasion of endometriosis.

Mast Cell Proteases Promote Diverse Effects on the Plasminogen Activation System and Wound Healing in A549 Alveolar Epithelial Cells.

Tissue damage, epithelial alterations, and intraepithelial presence of mast cells (MCs) are characteristics of asthma pathogenesis. Increased alveolar infiltration of MC populations has also been identified as a feature of asthma and other chronic respiratory diseases. The asthma associated receptor, urokinase plasminogen activator receptor (uPAR), has been shown to regulate bronchial epithelial repair responses. However, the impact of MC tryptase and chymase on functional properties and expression of uPAR in alveolar epithelial cells have not been fully investigated. Alveolar epithelial cell migration and wound healing were investigated using holographic live cell imaging of A549 cells in a wound scratch model post stimulation with tryptase or chymase. The expression of uPAR was investigated on the protein and gene level from cellular supernatants and in bronchoalveolar lavage fluid fractions from allergic asthmatics. We found that tryptase improved wound healing capacity, cellular migration and membrane bound uPAR expression. Chymase reduced gap closure capacity, cellular migration and membrane bound uPAR expression but increased soluble uPAR release. Our data suggest a dual regulatory response from the MC proteases in events related to uPAR expression and wound healing which could be important features in asthmatic disease.

Soluble Urokinase Plasminogen Activator Receptor Levels and Outcomes in Patients with Heart Failure

BackgroundSoluble urokinase-type plasminogen activator receptor (suPAR) is a marker of immune activation and pathogenic factor for kidney disease shown to predict cardiovascular outcomes including heart failure (HF) in various populations. We characterized suPAR levels in patients with HF and compared its ability to discriminate risk to that of B-type natriuretic peptide (BNP). Methods and ResultsWe measured plasma suPAR and BNP levels in 3,437 patients undergoing coronary angiogram and followed for a median of 6.2 years. We performed survival analyses for the following outcomes: all-cause death, cardiovascular death, and hospitalization for HF. We then assessed suPAR's ability to discriminate risk for the aforementioned outcomes. We identified 1116 patients with HF (age 65±12, 67.2% male, 20.0% Black, 67% with reduced ejection fraction). The median suPAR level was higher in HF compared to those without HF (3370 [IQR 2610-4371] vs. 2880 [IQR 2270-3670] pg/mL, respectively, P<0.001). In patients with HF, suPAR levels (log-base 2) were associated with outcomes including all-cause death (adjusted hazard ratio aHR 2.30, 95%CI[1.90-2.77]), cardiovascular death (aHR 2.33 95%CI[1.81-2.99]) and HF hospitalization (aHR 1.96, 95%CI[1.06-1.25]) independently of clinical characteristics and BNP levels. The association persisted across subgroups and did not differ between patients with reduced or preserved ejection fraction, or those with ischemic or non-ischemic cardiomyopathy. Addition of suPAR to a model including BNP levels significantly improved the C-statistic for death (Δ0.027), cardiovascular death (Δ0.017) and hospitalization for HF (Δ0.017). ConclusionsSuPAR levels are higher in HF compared to non-HF, are strongly predictive of outcomes, and combined with BNP, significantly improved risk prediction. Lay Summary•Soluble urokinase plasminogen activator receptor (suPAR) is a circulating protein of immune origin notorious for its involvement in kidney disease, and which levels have been found to predict the onset of heart failure.•Given the pathophysiologic link between heart failure and kidney disease, we sought to examine suPAR levels in patients with heart failure.•We measured suPAR in 1116 patients with heart failure, and found that levels were strongly predictive of outcomes independently of risk factors and above and beyond the myocardial marker BNP. SuPAR level may be useful as an adjunctive measure for risk stratification of patients with heart failure.

Assessment of clinical utility and predictive potential of pre-chemotherapy soluble urokinase plasminogen activator receptor: Observational single center study.

Alteration of urokinase plasminogen activator receptor (uPAR) in neoplasms is a prerequisite for invasiveness and metastatic ability. In the present study, we aimed to evaluate the relationship of pre-chemotherapy soluble uPAR (suPAR) with the odds for metastasis, lack of disease control, and its predictive ability for progression-free survival (PFS). Baseline plasma suPAR levels were measured by ELISA in 89 patients with various cancers prior to initiation of systemic treatment. Patients were followed prospectively until metastatic progression or death. TCGA Pan-Cancer dataset was mined for available RNAseq expression data of the PLAUR gene in patients with breast, colon, and lung cancer, and therelevant genomic and clinical data were extracted for further analysis. Pre-chemotherapy suPAR levels were significantly associated with white blood cell counts and fibrinogen and were significantly elevated both in patients with metastatic disease and in patients with progression. Increasing suPAR was significantly associated with odds for progression in the prespecified multivariate analysis (odds ratio 2.47, 95% confidence interval 1.3 - 5.11). In univariate Cox regression, suPAR was predictive of shortened progression-free survival (PFS) (hazard ratio 1.065, 95% confidence interval 1.002 - 1.13; p = 0.041). There was a trend towards shortened PFS in patients with higher baseline suPAR levels (cutoff 8.1 ng/mL). In the TCGA lung cancer cohort, PLAUR mRNA expression was significantly associated with shortened PFS in both univariate and multivariate analyses. High PLAUR gene expression conferred significant survival disadvantage only in patients with colon and lung cancer. SuPAR may bear predictive potential for adverse outcomes in cancer, but its utility as a biomarker seems to be more pronounced in cancers with associated inflammatory state.

Bioinformatics Analysis of Inflammation Gene Signature in Indicating Cholangiocarcinoma Prognosis

Aim We studied inflammatory response-related genes in cholangiocarcinoma by bioinformatics analysis. Methods The expression profiles and clinical information of cholangiocarcinoma patients were downloaded from the TCGA cohort and the Gene Expression Omnibus. The greatest absolute shrinking and selecting operator Cox analyses were utilized to build a multigene predictive signature. Results An inflammation response-related gene profile was generated using LASSO-Cox regression analysis of Homo sapiens bestrophin 1 (BEST1), Chemokine (C–C motif) ligand 2 (CCL2), and plasminogen activator, urokinase receptor (PLAUR). Individuals in the highest category had a significantly lower overall survival time than those from the low-risk group. A receiver operating curve analysis was used to demonstrate the predictive ability of the predictive gene signature. Through multivariate Cox analysis, the risk score was discovered to be a predictor of overall survival (OS). According to functional assessments, the immunological state and milieu of the two risk areas were significantly different. The expression levels of predictive genes were found to be strongly linked to the sensitivity of cancer cells to antitumor therapy. Conclusion A new signature made up of three respective response-relevant genes is found to be a promising indicator of prognosis by influencing the immune condition and tumor microenvironment.

Growth differentiation factor-15 as a biomarker of atherosclerotic coronary plaque: Value in people living with and without HIV.

BackgroundIncreased rates of cardiovascular diseases (CVD) and larger subclinical high-risk coronary plaques in coronary CT angiography have been observed in people living with HIV (PLWH) treated with antiretroviral therapy (ART) compared to HIV-uninfected people. Growth differentiation factor-15 (GDF-15) is a cytokine emerging as an optimal marker for CVD in the general population.MethodsWe cross-sectionally analyzed plasma of 95 PLWH on ART and 52 controls. We measured GDF-15, fibroblast growth factor-21 (FGF-21), glucagon-like peptide-2 (GLP-2), soluble urokinase plasminogen activator receptor (suPAR), CRP, and anti-CMV and anti-EBV IgG levels. All participants had no clinical CVD and underwent coronary CT angiography with the 3D reconstruction of coronary artery atherosclerotic plaques. Total plaque volume (TPV) and low attenuation plaque volume (LAPV, defined as density <30 Hounsfield Units) were calculated (mm3).ResultsIn both PLWH and controls, GDF-15 levels were increased in participants with presence of coronary plaque vs. without (p = 0.04 and p < 0.001, respectively) and correlated with TPV (r = 0.27, p = 0.009 and r = 0.62, p < 0.001, respectively) and LAPV (r = 0.28, p = 0.008, r = 0.60, p < 0.001, respectively). However, in a multivariate model, GDF-15 was independently associated with LAPV in controls only (adjusted OR 35.1, p = 0.04) and not in PLWH, mainly due to confounding by smoking. Other markers were not independently associated with plaque volume, except for anti-EBV IgGs in controls (adjusted OR 3.51, p = 0.02).ConclusionIn PLWH, GDF-15 and smoking seemed to synergistically contribute to coronary plaque volume. Conversely, increased GDF-15 levels were associated with the presence of coronary artery plaques in people without HIV, independently of CV risk factors.