To investigate whether obesity induces a leptin–Notch signaling axis in breast cancer (BC), leptin-induced Notch was determined in human MCF-7 and MDA-MB231 and mouse E0771 cells and, in E0771-BC hosted by syngeneic lean and diet-induced-obesity (DIO) C57BL/6J female mice. Lean and DIO-mice were treated for three weeks with leptin inhibitor (PEG-LPrA2) one week after the inoculation of E0771 cells. Leptin induced Notch1, 3 and 4 in BC cells, but Notch2 expression showed opposite pattern in MCF-7 compared to MDA-MB231 cells. Notch loss-of-function [DAPT and dominant negative (R218H) RBP-Jk (CSL/CBF1)] showed that a functional leptin-Notch signaling axis was involved in the proliferation and migration of E0771 cells. E0771-BC onset was affected by obesity [lean mice: 7/10 (70%) vs DIO-mice: 11/12 (92%); Pearson Chi2: P=0.06]. PEG-LPrA2 significantly reduced BC growth [untreated: 19/42; (45%) vs treated: 8/42 (19%); Pearson Chi2: p=0.008]. PEG-LPrA2 did not influence the caloric intake of mice, but increased carcass and/or body weights of lean and DIO-mice inoculated with E0771 cells, which could be related to the improvement of health conditions (less aggressive disease). Importantly, BC from obese mice had higher levels of Notch3, JAG-1 and survivin than lean mice. Inhibition of leptin signaling reduced protein levels of Notch (NICD1, NICD4, Notch3, JAG1 and survivin) and significantly decreased mRNA expression of Notch receptors, ligands, and targets. PEG-LPrA's effects were more prominent in DIO-mice. Present data suggest that leptin induces Notch, which could be involved in the reported higher incidence and aggressiveness and, poor prognosis of BC in obese patients.

Background The mortality of severe malaria [cerebral malaria (CM), severe malaria anemia (SMA), acute lung injury (ALI) and acute respiratory distress syndrome (ARDS)] remains high despite the availability associated with adequate treatments. Recent studies in our laboratory and others have revealed a hitherto unknown correlation between chemokine CXCL10/CXCR3, Heme/HO-1 and STAT3 and cerebral malaria severity and mortality. Although Heme/HO-1 and CXCL10/CXCR3 interactions are directly involved in the pathogenesis of CM and fatal disease, the mechanism dictating how Heme/HO-1 and CXCL10/CXCR3 are expressed and regulated under these conditions is still unknown. We therefore tested the hypothesis that these factors share common signaling pathways and may be mutually regulated. Methods We first clarified the roles of Heme/HO-1, CXCL10/CXCR3 and STAT3 in CM pathogenesis utilizing a well established experimental cerebral malaria mouse (ECM, P. berghei ANKA) model. Then, we further determined the mechanisms how STAT3 regulates HO-1 and CXCL10 as well as mutual regulation among them in CRL-2581, a murine endothelial cell line. Results The results demonstrate that (1) STAT3 is activated by P. berghei ANKA (PBA) infection in vivo and Heme in vitro . (2) Heme up-regulates HO-1 and CXCL10 production through STAT3 pathway, and regulates CXCL10 at the transcriptional level in vitro . (3) HO-1 transcription is positively regulated by CXCL10. (4) HO-1 regulates STAT3 signaling. Conclusion Our data indicate that Heme/HO-1, CXCL10/CXCR3 and STAT3 molecules as well as related signaling pathways play very important roles in the pathogenesis of severe malaria. We conclude that these factors are mutually regulated and provide new opportunities to develop potential novel therapeutic targets that could be used to supplement traditional prophylactics and treatments for malaria and improve clinical outcomes while reducing malaria mortality. Our ultimate goal is to develop novel therapies targeting Heme or CXCL10-related biological signaling molecules associated with development of fatal malaria.

Background: Lupus nephritis (LN) occurs in up to 40% of patients with systemic lupus erythematosus (SLE). Reliable biomarkers of kidney damage are needed to identify SLE patients at risk to develop LN in order to improve screening, treat earlier, and halt progression to kidney failure. Novel biomarkers of extracellular matrix remodeling were evaluated as markers of kidney fibrosis and disease activity in LN patients. Methods: Biomarkers of the interstitial collagen type III (PRO-C3) and type VI (PRO-C6) formation as well as of collagen type III (C3M) degradation were evaluated in the serum and urine of 40 patients with LN, 20 SLE patients without LN, 20 healthy controls and 10 biopsy controls (histological kidney inflammation/damage without SLE). Their association with histological markers of interstitial fibrosis and tubular atrophy, with inflammatory cell infiltration and with disease activity and chronicity in the LN patients was assessed. Results: Despite PRO-C3 (serum) and PRO-C6 (serum and urine) were significantly elevated in LN patients compared to healthy controls, they were not able to separate the LN from the SLE patients. C3M (urine) levels were not different in the LN group compared to the others. C3M (urine) strongly correlated and PRO-C6 (serum and urine) inversely correlated with kidney function (eGFR). The biomarkers of interstitial collagen turnover PRO-C6 (serum) and C3M (urine) correlated with histological markers of interstitial fibrosis, tubular atrophy, and monocyte infiltration. Conclusions: Non-invasive collagen turnover biomarkers are promising tools to identify SLE patients with kidney histological modifications.

The endocannabinoid system is an emerging immunomodulatory network that may be involved in AERD. This is the first study of CB2R in sinonasal disease, showing significantly increased transcription in nasal polyps from subjects with AERD. Additional study is warranted to further evaluate the contribution and therapeutic potential of this novel finding in chronic rhinosinusitis.

No abstract

No abstract

ObjectiveLupus nephritis (LN) is diagnosed by biopsy, but longitudinal monitoring assessment methods are needed. Here, in this preliminary and hypothesis-generating study, we evaluate the potential for using urine proteomics as a non-invasive method to monitor disease activity and damage. Urinary biomarkers were identified and used to develop two novel algorithms that were used to predict LN activity and chronicity.MethodsBaseline urine samples were collected for four cohorts (healthy donors (HDs, n=18), LN (n=42), SLE (n=17) or non-LN kidney disease biopsy control (n=9)), and over 1 year for patients with LN (n=42). Baseline kidney biopsies were available for the LN (n=46) and biopsy control groups (n=9). High-throughput proteomics platforms were used to identify urinary analytes ≥1.5 SD from HD means, which were subjected to stepwise, univariate and multivariate logistic regression modelling to develop predictive algorithms for National Institutes of Health Activity Index (NIH-AI)/National Institutes of Health Chronicity Index (NIH-CI) scores. Kidney biopsies were analysed for macrophage and neutrophil markers using immunohistochemistry (IHC).ResultsIn total, 112 urine analytes were identified from LN, SLE and biopsy control patients as both quantifiable and overexpressed compared with HDs. Regression analysis identified proteins associated with the NIH-AI (n=30) and NIH-CI (n=26), with four analytes common to both groups, demonstrating a difference in the mechanisms associated with NIH-AI and NIH-CI. Pathway analysis of the NIH-AI and NIH-CI analytes identified granulocyte-associated and macrophage-associated pathways, and the presence of these cells was confirmed by IHC in kidney biopsies. Four markers each for the NIH-AI and NIH-CI were identified and used in the predictive algorithms. The NIH-AI algorithm sensitivity and specificity were both 93% with a false-positive rate (FPR) of 7%. The NIH-CI algorithm sensitivity was 88%, specificity 96% and FPR 4%. The accuracy for both models was 93%.ConclusionsLongitudinal predictions suggested that patients with baseline NIH-AI scores of ≥8 were most sensitive to improvement over 6–12 months. Viable approaches such as this may enable the use of urine samples to monitor LN over time.

and Wome. RATIONALE: Nasal fibroblasts from subjects with nasal polyps display impaired production of prostaglandin (PG) E2 compared with controls for incompletely understood reasons. PGE2 produced by human amnion fibroblasts can amplify cyclooxygenase (COX)2 expression and PGE2 synthesis via EP2/EP4 with the cyclic adenosine 3', 5'-monophosphate (cAMP) and Protein kinase A (PKA) pathway. Whether a PGE2-dependent feed-forward loop regulates COX-2 in nasal fibroblasts is not clarified. METHODS: RNAseq was performed on a subset of freshly isolated polyp fibroblasts from patients with (n 5 3) and without (n 5 6) aspirinexacerbated respiratory disease (AERD), as well as non-polyp controls. Nasal fibroblasts cultured from 13 patients undergoing polyp surgery at Brigham and Women's hospital were stimulated with IL-1b 1ng/ml in the presence or absence of PKA inhibitor (H89), a nonselective cyclooxygenase (COX) inhibitor (indomethacin), a COX-1 inhibitor (SC560), a COX-2 inhibitor (SC236), EP2 inhibitor (PF04418948), or EP4 inhibitor (ONO). PGE2 levels and cAMP accumulation were measured by ELISA kit (Cayman). RESULTS: Human nasal fibroblasts produced PGE2 at 6-24 h after the IL-1b stimulation accompanied with expressions of COX2 and mPGES-1. cAMP accumulated most at 6h after the stimulation. H89, indomethacin, SC560, and SC236 reduced IL-1b induced PGE2 levels significantly. Additionally, indomethacin and SC236 could inhibit cAMP accumulation completely. PRKACB levels were markedly reduced in freshly sorted AERD fibroblasts compared with non-polyp controls. CONCLUSIONS: Our results supported the regulation of PGE2 production through feed-forward loop in nasal fibroblasts. Intrinsically diminished EP2/PKA/COX2 pathway may contribute to impairment of PGE2 production in nasal polyps, especially in AERD.