Targeting Inflammatory Pathways Research Articles

Repurposing of Antifungal Drug Flucytosine/Flucytosine Cocrystals for Anticancer Activity against Prostate Cancer Targeting Apoptosis and Inflammatory Signaling Pathways.

This study aimed to repurpose the antifungal drug flucytosine (FCN) for anticancer activity together with cocrystals of nutraceutical coformers sinapic acid (SNP) and syringic acid (SYA). The cocrystal screening experiments with SNP resulted in three cocrystal hydrate forms in which two are polymorphs, namely, FCN-SNP F-I and FCN-SNP F-II, and the third one with different stoichiometry in the asymmetric unit (1:2:1 ratio of FCN:SNP:H2O, FCN-SNP F-III). Cocrystallization with SYA resulted in two hydrated cocrystal polymorphs, namely, FCN-SYA F-I and FCN-SYA F-II. All the cocrystal polymorphs were obtained concomitantly during the slow evaporation method, and one of the polymorphs of each system was produced in bulk by the slurry method. The interaction energy and lattice energies of all cocrystal polymorphs were established using solid-state DFT calculations, and the outcomes correlated with the experimental results. Further, the in vitro cytotoxic activity of the cocrystals was determined against DU145 prostate cancer and the results showed that the FCN-based cocrystals (FCN-SNP F-III and FCN-SYA F-I) have excellent growth inhibitory activity at lower concentrations compared with parent FCN molecules. The prepared cocrystals induce apoptosis by generating oxidative stress and causing nuclear damage in prostate cancer cells. The Western blot analysis also depicted that the cocrystals downregulate the inflammatory markers such as NLRP3 and caspase-1 and upregulate the intrinsic apoptosis signaling pathway marker proteins, such as Bax, p53, and caspase-3. These findings suggest that the antifungal drug FCN can be repurposed for anticancer activity.

Novel insights into the anti-asthmatic effect of Raphanus sativus L. (Raphani Semen): Targeting immune cells, inflammatory pathways and oxidative stress markers

Ethnopharmacological relevanceRaphanus sativus L. is a well-known medicinal plant with traditional therapeutic applications in various common ailments including inflammation and asthma. Aims of the studyThis study aimed to evaluate the chemical composition and anti-asthmatic potential of the hydro-methanolic extract of the leaves of R. sativus L. (Rs.Cr) using various in vitro and in vivo investigations. Materials and methodsThe Rs.Cr was subjected to preliminary phytochemical analysis and HPLC profiling. The safety was assessed through oral acute toxicity tests in mice. The antiasthmatic effect of the extract was studied using milk-induced leukocytosis and ovalbumin (OVA)-induced allergic asthma models established in mice. While mast cell degranulation and passive paw anaphylaxis models were established in rats. Moreover, effect of the extract was studied on various oxidative and inflammatory makers. The antioxidant effect of the extract was also studied by in vitro DPPH method. ResultsThe HPLC profiling of Rs.Cr showed the presence of important polyphenols in a considerable quantity. In toxicity evaluation, Rs.Cr showed no sign of morbidity or mortality with LD50 < 2000 mg/kg. The extract revealed significant mast cell disruption in a dose-dependent manner compared to the intoxicated group. Similarly, treatment with Rs.Cr and dexamethasone significantly (p < 0.001) reduced paw edema volume. Subcutaneous injection of milk at a dose of 4 mL/kg, after 24 h of its administration, showed an increase in the leukocyte count in the intoxicated group. Similarly, mice treated with dexamethasone and Rs.Cr respectively showed a significant decrease in leukocytes and eosinophils count in the ovalbumin-induced allergic asthma model. The extract presented a significant (p˂0.001) alleviative effect on the levels of SOD and GSH, MDA, IL-4, IL-5, and IL-13 in a dose-dependent manner as compared to the intoxicated group. Furthermore, the histological evaluation also revealed a notable decrease in inflammatory and goblet cell count with reduced mucus production. ConclusionThe current study highlights mechanism-based novel insights into the anti-asthmatic potential of R. sativus that also strongly supports its traditional use in asthma.

A Bispecific, Tetravalent Antibody Targeting Inflammatory and Pruritogenic Pathways in Atopic Dermatitis

Inhibition of IL-4/IL-13 signaling has dramatically improved the treatment of atopic dermatitis (AD). However, in many patients, clinical responses are slow to develop and remain modest. Indeed, some symptoms of AD are dependent on IL-31, which is only partially reduced by IL-4/IL-13 inhibition. Thus, there is an unmet need for AD treatments that concomitantly block IL-4/IL-13 and IL-31 pathways. We engineered NM26-2198, a bispecific tetravalent antibody designed to accomplish this task. In reporter cell lines, NM26-2198 concomitantly inhibited IL-4/IL-13 and IL-31 signaling with a potency comparable with that of the combination of an anti-IL-4Rα antibody (dupilumab) and an anti-IL-31 antibody (BMS-981164). In human PBMCs, NM26-2198 inhibited IL-4-induced upregulation of CD23, demonstrating functional binding to FcγRII (CD32). NM26-2198 also inhibited the secretion of the AD biomarker thymus and activation-regulated chemokine (TARC) in blood samples from healthy human donors. In male cynomolgus monkeys, NM26-2198 exhibited favorable pharmacokinetics and significantly inhibited IL-31-induced scratching at a dose of 30 mg/kg. In a repeat-dose, good laboratory practice toxicology study in cynomolgus monkeys, no adverse effects of NM26-2198 were observed at a weekly dose of 125 mg/kg. Together, these results justify the clinical investigation of NM26-2198 as a treatment for moderate-to-severe AD.

Tasquinimod Improves Erythropoiesis and Mitigates Bone Loss in Myelodysplastic Mice

Myelodysplastic neoplasms (MDS) are characterized by ineffective hematopoiesis, peripheral cytopenia, and a risk of transforming into acute myeloid leukemia (AML). Interestingly, MDS is also associated with an increased prevalence of osteoporosis. Studies have suggested that dysregulated innate immune responses and a proinflammatory bone marrow (BM) microenvironment play significant roles in the pathogenesis of MDS. Targeting inflammatory pathways, especially in low-risk MDS patients, presents a potential strategy to modulate the progression of the disease. One such candidate is Tasquinimod (TASQ, ABR-215050, Active Biotech AB), a small-molecule oral inhibitor and second-generation quinoline-3-carboxamide compound. TASQ binds to the inflammatory protein S100A9 and inhibits its interaction with the proinflammatory toll-like receptor 4, thereby displaying both immunomodulatory and antitumor properties. Our in vitro data provided evidence for a rescuing effect of TASQ on inflammatory signal-primed MDS stromal cells, leading to enhanced hematopoietic support. This study aims to validate these findings using a transgenic MDS mouse model expressing the NUP98/HOXD13 (NHD13) fusion protein in hematopoietic cells. NHD13 mice, generated on a C57BL/6J background, were obtained from Jackson Laboratory. TASQ was administered orally by adding it to the drinking water at a dose of 30 mg/kg. Both transgenic and littermate control (WT) mice (12 weeks old, n=5-10 per group) were treated with or without TASQ for 12 weeks. Throughout the experiment, peripheral blood cell counts were measured every 4 weeks using Sysmex analysis. In the final analysis, primary BM cells were isolated from the femora, and the number of lineage-negative, sca-1-positive, and c-kit-positive (LSK) cells, as well as erythroid differentiation, were assessed using flow cytometry. Additionally, bone volume, trabecular number, and thickness were analyzed in femora using µCT. Transgenic NHD13 mice displayed MDS-like characteristics, including peripheral cytopenias, dysplasia in blood and BM cells, and microstructural abnormalities in the bone microenvironment. As expected, hemoglobin levels (Hb) and red blood cell (RBC) counts were lower in NHD13 mice compared to WT mice [Hb: 8.9 vs. 9.5 mmol/L; RBC: 9.5 vs. 10.3x1012/L] and showed a consistent decrease in the untreated group from 12 to 24 weeks. Notably, treatment with TASQ significantly improved red blood cell counts in NHD13 mice [+14%; p&amp;lt;0.01] after 24 weeks, while having no effect on WT mice. White blood cells were significantly decreased in WT mice after 4 weeks of TASQ treatment [-19%; p&amp;lt;0.001] until the end of the experiment [-36%; p&amp;lt;0.001]. In contrast, in transgenic mice, the lower white blood cell counts [-76%; p&amp;lt;0.001] were not further diminished. Interestingly, TASQ led to an elevation in neutrophil counts. The platelet count was not affected by the treatment in both transgenic and WT mice. Using flow cytometry, an increase in the total number of LSK cells was detected in the BM of WT mice after TASQ treatment. However, no increase was observed in NHD13 mice, which already had lower LSK numbers compared to WT controls. Late-stage erythropoiesis was slightly increased in both WT and transgenic mice after TASQ treatment, whereas early erythroid differentiation was unaffected or even decreased by TASQ. Moreover, BM cells were used for colony-forming assays ex vivo. NHD13 mice had a lower erythroid differentiation potential compared to WT mice, which was improved after TASQ treatment. To assess the potential influence of the microenvironment, adherent BM cells were assayed for their CFU-F forming capacity. BM cells of transgenic mice exhibited a slightly lower CFU-F forming capacity compared to WT mice, which was partially reversed by TASQ treatment. Finally, the bone microarchitecture was analyzed. NHD13 mice showed a lower bone volume [-43%; p&amp;lt;0.01] compared to WT mice. Notably, treatment with TASQ abolished the bone loss in NHD13 mice by increasing the trabecular number [+35% p&amp;lt;0.001], whereas the trabecular thickness was not affected. In conclusion, this study provides the first evidence for an in vivo effect of TASQ in a murine model of MDS, suggesting improved erythropoiesis and positive effects on the bone phenotype. These results warrant further investigations of TASQ in human trials, particularly in MDS patients in whom anemia and bone loss often coexist.

The p65-LOC727924-miR-26a/KPNA3-p65 regulatory loop mediates vasoactive intestinal peptide effects on osteoarthritis chondrocytes.

Loss and dysfunction of articular chondrocytes, which disrupt the homeostasis of extracellular matrix formation and breakdown, promote the onset of osteoarthritis (OA). Targeting inflammatory pathways is an important therapeutic strategy for OA. Vasoactive intestinal peptide (VIP) is an immunosuppressive neuropeptide with potent anti-inflammatory effects; however, its role and mechanism in OA remain unclear. In this study, microarray expression profiling from the Gene Expression Omnibus database and integrative bioinformatics analyses were performed to identify differentially expressed lncRNAs in OA samples. qRT-PCR validation of the top ten different expressed lncRNAs indicated that the expression level of intergenic non-protein coding RNA 2203 (LINC02203, also named LOC727924) was the highest in OA cartilage compared to normal cartilage. Hence, the LOC727924 function was further investigated. LOC727924 was upregulated in OA chondrocytes, with a dominant sub-localization in the cytoplasm. In OA chondrocytes, LOC727924 knockdown boosted cell viability, suppressed cell apoptosis, reactive oxygen species (ROS) accumulation, increased aggrecan and collagen II, decreased matrix metallopeptidase (MMP)-3/13 and ADAM metallopeptidase with thrombospondin type 1 motif (ADAMTS)-4/5 levels, and reduced the levels of tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL-1β), and interleukin 6 (IL-6). LOC727924 could interact with the microRNA 26a (miR-26a)/ karyopherin subunit alpha 3 (KPNA3) axis by competitively targeting miR-26a for KPNA3 binding, therefore down-regulating miR-26a and upregulating KPNA3; in OA chondrocytes, miR-26a inhibition partially abolished LOC727924 knockdown effects on chondrocytes. miR-26a inhibited the nuclear translocation of p65 through targeting KPNA3 and p65 transcriptionally activated LOC727924, forming a p65-LOC727924-miR-26a/KPNA3-p65 regulatory loop to modulate OA chondrocyte phenotypes. In vitro, VIP improved OA chondrocyte proliferation and functions, down-regulated LOC727924, KPNA3, and p65 expression, and upregulated miR-26a expression; in vivo, VIP ameliorated destabilization of the medial meniscus (DMM)-induced damages on the mouse knee joint, down-regulated KPNA3, inhibited the nuclear translocation of p65. In conclusion, the p65-LOC727924-miR-26a/KPNA3-p65 regulatory loop modulates OA chondrocyte apoptosis, ROS accumulation, extracellular matrix (ECM) deposition, and inflammatory response in vitro and OA development in vivo, being one of the mechanisms mediating VIP ameliorating OA.

Hepatoprotective effects of Gynura procumbens against thioacetamide-induced cirrhosis in rats: Targeting inflammatory and oxidative stress signalling pathways

Gynura procumbens is an edible flowering plant that has been utilized as traditional therapy for numerous diseases. The current experiment investigates the hepatoprotective potentials of the ethanol extract of Gynura procumbens leaf (EEGPL) against thioacetamide (TAA)-induced liver cirrhosis in rats. Thirty Sprague Dawley rats were randomly divided into 5 clusters: A, rats received orally 10% Tween 80 and intraperitoneal (i.p) inoculation of sterile distal water; B, rats received orally10% Tween 80; C, rats received orally daily 50 mg/kg of silymarin, while groups; D and E, rats received orally daily doses of 200 and 400 mg/kg of EEGPL, respectively. Furthermore, B-E clusters received 200 mg/kg thioacetamide (i.p) three times a week for 60 days.The liver gross morphology of rats that received only TAA (B) revealed irregular rough surface layers compared to smoother livers of rats that received EEGPL. Histopathology of group B revealed clear hepatic necrosis and fibrous connective tissue, which were significantly reduced in C-E groups. EEGPL treatment caused a significant down-regulation of PCNA and α-SMA protein expressions. Antioxidant (SOD and CAT) enzymes in hepatic homogeneity were meaningfully lower, and MDA levels were significantly higher in TAA controls compared to those of C-E groups. Moreover, EEGPL treatment caused a reduction of TNF-α and IL-6 and increased expression of IL-10 cytokines. Therefore, the hepatoprotective potentials of EEGPL might be contributed to its modulation of detoxification enzymes, anti-inflammatory, and antioxidant activities.

Stairway to Heaven: Do head-to-head Trials Indicate a Need for Definite Criteria for Choosing Biologic Drugs in Rheumatoid Arthritis?

Rheumatoid arthritis (RA) is a chronic disease characterized by joint and systemic involvement that develops with different pathogenetic mechanisms. Treatment of the disease is undertaken with disease-modifying anti-rheumatic drugs (DMARDs). The mechanisms of action of conventional DMARDs generally are based on the inhibition of T and B-cells in the immune system. In recent years, biologic and targeted smart molecules have been used in the treatment of RA. Targeting different cytokines and inflammatory pathways, these drugs have ushered in a new era in RA treatment. The efficacy of these drugs has been demonstrated in many studies; and in the postmarketing period, that is, as the patients who use them say, they are like a "stairway to heaven". However, as every "road to heaven" is challenging and "thorny", the efficacy and reliability of these drugs and whether any one of them is superior to the others, remains a matter of debate. However, the use of biologic drugs with or without cDMARDs, the preference for original vs. biosimilar molecules, and discontinuation of the drugs after achieving sustained remission are other questions that need to be explored. When it comes to the choice of biological drugs by rheumatologists, it is not yet clear on which criteria they base their choices on. Due to the limited comparative studies of these biological drugs, the subjective criteria of the physician gains importance. The selection of these drugs, however, should be based on objective criteria such as efficacy, safety, superiority over each other, and cost. In other words, the determinant of the "path to heaven" should be based on objective criteria and recommendations according to the scientific data generated by controlledprospective studies, not on the initiative of a single physician. In this review, a head-to-head comparison of biological drugs used in the treatment of RA, their efficacy, safety, and which are superior are discussed in light of recent literature data.

Astrocytes as a target for therapeutic strategies in epilepsy: current insights.

Astrocytes are specialized non-neuronal glial cells of the central nervous system, contributing to neuronal excitability and synaptic transmission (gliotransmission). Astrocytes play a key roles in epileptogenesis and seizure generation. Epilepsy, as a chronic disorder characterized by neuronal hyperexcitation and hypersynchronization, is accompanied by substantial disturbances of glial cells and impairment of astrocytic functions and neuronal signaling. Anti-seizure drugs that provide symptomatic control of seizures primarily target neural activity. In epileptic patients with inadequate control of seizures with available anti-seizure drugs, novel therapeutic candidates are needed. These candidates should treat epilepsy with anti-epileptogenic and disease-modifying effects. Evidence from human and animal studies shows that astrocytes have value for developing new anti-seizure and anti-epileptogenic drugs. In this review, we present the key functions of astrocytes contributing to neuronal hyperexcitability and synaptic activity following an etiology-based approach. We analyze the role of astrocytes in both development (epileptogenesis) and generation of seizures (ictogenesis). Several promising new strategies that attempted to modify astroglial functions for treating epilepsy are being developed: (1) selective targeting of glia-related molecular mechanisms of glutamate transport; (2) modulation of tonic GABA release from astrocytes; (3) gliotransmission; (4) targeting the astrocytic Kir4.1-BDNF system; (5) astrocytic Na+/K+/ATPase activity; (6) targeting DNA hypo- or hypermethylation of candidate genes in astrocytes; (7) targeting astrocytic gap junction regulators; (8) targeting astrocytic adenosine kinase (the major adenosine-metabolizing enzyme); and (9) targeting microglia-astrocyte communication and inflammatory pathways. Novel disease-modifying therapeutic strategies have now been developed, such as astroglia-targeted gene therapy with a broad spectrum of genetic constructs to target astroglial cells.

Synthesis and In Vitro Biological Evaluation of p-Carborane-Based Di-tert-butylphenol Analogs.

Targeting inflammatory mediators and related signaling pathways may offer a rational strategy for the treatment of cancer. The incorporation of metabolically stable, sterically demanding, and hydrophobic carboranes in dual cycloxygenase-2 (COX-2)/5-lipoxygenase (5-LO) inhibitors that are key enzymes in the biosynthesis of eicosanoids is a promising approach. The di-tert-butylphenol derivatives R-830, S-2474, KME-4, and E-5110 represent potent dual COX-2/5-LO inhibitors. The incorporation of p-carborane and further substitution of the p-position resulted in four carborane-based di-tert-butylphenol analogs that showed no or weak COX inhibition but high 5-LO inhibitory activities in vitro. Cell viability studies on five human cancer cell lines revealed that the p-carborane analogs R-830-Cb, S-2474-Cb, KME-4-Cb, and E-5110-Cb exhibited lower anticancer activity compared to the related di-tert-butylphenols. Interestingly, R-830-Cb did not affect the viability of primary cells and suppressed HCT116 cell proliferation more potently than its carbon-based R-830 counterpart. Considering all the advantages of boron cluster incorporation for enhancement of drug biostability, selectivity, and availability of drugs, R-830-Cb can be tested in further mechanistic and in vivo studies.

Autism Spectrum Disorders: A Recent Update on Targeting Inflammatory Pathways with Natural Anti-Inflammatory Agents.

Autism spectrum disorder (ASD) is a heterogeneous category of developmental psychiatric disorders which is characterized by inadequate social interaction, less communication, and repetitive phenotype behavior. ASD is comorbid with various types of disorders. The reported prevalence is 1% in the United Kingdom, 1.5% in the United States, and ~0.2% in India at present. The natural anti-inflammatory agents on brain development are linked to interaction with many types of inflammatory pathways affected by genetic, epigenetic, and environmental variables. Inflammatory targeting pathways have already been linked to ASD. However, these routes are diluted, and new strategies are being developed in natural anti-inflammatory medicines to treat ASD. This review summarizes the numerous preclinical and clinical studies having potential protective effects and natural anti-inflammatory agents on the developing brain during pregnancy. Inflammation during pregnancy activates the maternal infection that likely leads to the development of neuropsychiatric disorders in the offspring. The inflammatory pathways have been an effective target for the subject of translational research studies on ASD.

Qu-Yu-Jie-Du Decoction Inhibits the Recurrence and Metastasis of Stage III Colorectal Cancer by Targeting Inflammatory Pathways: A Retrospective Clinical Study

Background: Colorectal Cancer (CRC) is among the leading causes of cancer-related mortality and morbidity worldwide. Qu-Yu-Jie-Du (QYJD) decoction 10 prevents enteritis and prolongs CRC survival time however, large11 scale clinical research and mechanism analysis are needed. Hence, our study aimed to confirm the effectiveness of QYJD decoction in 13 stage III CRC patients and its potential anticancer mechanisms.

Targeting Inflammatory Pathways to Reverse Immunosuppressive Tumor Microenvironment in Chronic Lymphocytic Leukemia

Targeting Inflammatory Pathways to Reverse Immunosuppressive Tumor Microenvironment in Chronic Lymphocytic Leukemia

Inflammatory molecules facilitate the development of docetaxel-resistant prostate cancer cells in vitro and in vivo.

Numerous molecular mechanisms have been found to contribute to docetaxel-induced resistance in prostate cancer (PCa). In this study, the changes in gene expression profiles of multidrug resistant PCa cells that were established in response to docetaxel were determined using microarray analysis. In addition to alterations in the expression of multidrug resistance-associated genes, the expression levels of multiple inflammatory molecules, in particular IL-6, significantly increased in resistant cells in vitro and in vivo, which further increased with the development of drug resistance following microarray, qRT-PCR and ELISA analysis. Compared with parental cells, resistant cells also presented with stronger activation of multiple IL-6-associated signaling pathways STAT1/3, NF-κB, and PI3K/AKT. Inactivation of IL-6 using a neutralizing antibody resulted in a slight effect on the sensitivity of resistant cells to docetaxel, while blockade of of STAT1/3, NF-κB, or PI3K/AKT signaling significantly resensitized resistant cells to docetaxel. Of note, simultaneous inactivation of IL-6 and STAT1/3, PI3K/AKT or NF-κB further enhanced the sensitivity of the resistant cells to docetaxel. Thus, inflammatory molecules, in particular IL-6, and IL-6-associated signaling pathways NF-κB, STAT1/3, and PI3K/AKT, are crucial mediators of the development of docetaxel-resistance in PCa. Targeting inflammatory molecules and signaling pathways could be a potential therapeutic option for the intervention of drug resistance in PCa.

YOSEMITE and RHINE: Phase 3 Randomized Clinical Trials of Faricimab for Diabetic Macular Edema: Study Design and Rationale.

PurposeFaricimab is a novel anti–angiopoietin-2 and anti–vascular endothelial growth factor (VEGF) bispecific antibody with high affinities and specificities for both VEGF and angiopoietin-2. It is postulated that targeting angiogenic factors and inflammatory pathways in addition to the VEGF pathway will increase treatment durability and improve outcomes. The phase 3 YOSEMITE (ClinicalTrials.gov identifier, NCT03622580) and RHINE (ClinicalTrials.gov identifier, NCT03622593) trials are designed to assess efficacy, safety, and durability of faricimab compared with aflibercept in patients with diabetic macular edema (DME). The trials evaluate a personalized treatment interval (PTI) approach to address heterogeneity in treatment response among patients with DME.DesignTwo identically designed, global, double-masked, randomized, controlled phase 3 trials (YOSEMITE and RHINE).ParticipantsAdults with center-involving DME secondary to type 1 or 2 diabetes mellitus.MethodsThese studies were designed to evaluate 3 treatment groups: faricimab 6.0 mg dosed either at fixed dosing every 8 weeks after initial treatment with 6 intravitreal doses at 4-week intervals, or faricimab 6.0 mg dosed according to PTI after initial treatment with 4 every-4-week doses, compared with aflibercept 2.0 mg dosed every 8 weeks after 5 initial every-4-week doses. The primary end point of the studies was change from baseline in best-corrected visual acuity at 1 year, averaged over weeks 48, 52, and 56. Secondary end points included anatomic, durability, and patient-reported outcomes. Safety outcomes included incidence and severity of ocular and nonocular adverse events. The PTI is a protocol-defined flexible regimen based on the treat-and-extend concept, which allowed up to every-16-week adjustable dosing based on objective and standardized criteria. The PTI design aimed to maximize therapeutic results while minimizing treatment burden.Main Outcome MeasuresWe describe the rationale for the study design and the novel PTI (up to every-16-week adjustable dosing) approach for treatment with faricimab.ResultsYOSEMITE and RHINE enrolled 940 and 951 patients, respectively. Results from each study will be reported separately.ConclusionsYOSEMITE and RHINE were the first registrational trials in retinal disease to incorporate an objective PTI regimen, allowing for up to every-16-week adjustable dosing with a dual angiopoietin-2 and VEGF-A inhibitor, faricimab 6.0 mg, for treatment of DME.

A pro-resolving phenotype underpins the anti-inflammatory effects of inorganic nitrate

Abstract Introduction Increasing evidence highlights the critical role of chronic inflammation in cardiovascular disease (CVD). Targeting inflammatory pathways in patients with CVD has been associated with improved CV function in pre-clinical (Gee, 2017), early clinical (Yndestad, 2006; Velmurugan 2013; Jones, 2016) and large phase III studies (Ridker, 2017). The resolution of inflammation is an active process and its failure has also been proposed to contribute to CVD progression. At least one mechanism thought to underlie this failure is dysfunction of the canonical pathway for anti-inflammatory nitric oxide (NO) production. Restoring NO through provision of inorganic nitrate (NO3-) and subsequent bioactivation via the non-canonical pathway may offer therapeutic benefit. Aim To test whether dietary NO3–derived NO accelerates resolution of inflammation. Methods Randomised, double-blind, placebo-controlled, parallel limb study of 8–10mmol dietary NO3- supplementation versus NO3–deplete placebo beetroot juice in 36 healthy male volunteers (NCT03183830). Using a cantharadin-induced skin blister model (Day, 2001), acute (24h) and chronic (72h)-phase blisters were harvested pre- and post-treatment. Blister exudate was analysed for leucocyte activation state (CD11b, CD62L, CD162) by flow cytometry and cytokine/chemokine composition by ELISA. Ozone chemiluminescence established NO3-/NO2- levels in key biological matrices: plasma, urine and saliva. Results 9.3mmol inorganic NO3- led to a significant rise (versus placebo, p&amp;lt;0.001) of NO3-/NO2- in plasma, saliva and urine NO2- (p&amp;lt;0.02). No differences were seen in blister volumes, cell counts or markers of systemic inflammation. Whilst no differences were seen in the proportions of cellular infiltrate in 24h blisters, there were significant reductions of neutrophil (p=0.017) and intermediate monocyte proportions (p=0.001) and cellular adhesion molecules across inflammatory, intermediate and resolving monocytes at 72h (Figure 1). Generally, no differences in blister cytokine/chemokine profile was evident except for borderline significant suppression of TNFα at 24hrs with dietary NO3- treatment (P=0.057). Conclusion Whilst dietary inorganic NO3- does not impair the essential host defence response it does accelerate resolution: enhanced pro- to anti-inflammatory monocyte subtype switching and curtailed neutrophil recruitment, likely via attenuated TNFα production. These actions offer a novel, easy to administer, approach to influence inflammatory responses without impairing host defence. Funding Acknowledgement Type of funding sources: Public Institution(s). Main funding source(s): Derek Willoughby Trust and British Heart Foundation

Targeting inflammation in atherosclerosis - from experimental insights to the clinic.

Atherosclerosis, a dominant and growing cause of death and disability worldwide, involves inflammation from its inception to the emergence of complications. Targeting inflammatory pathways could therefore provide a promising new avenue to prevent and treat atherosclerosis. Indeed, clinical studies have now demonstrated unequivocally that modulation of inflammation can forestall the clinical complications of atherosclerosis. This progress pinpoints the need for preclinical investigations to refine strategies for combatting inflammation in the human disease. In this Review, we consider a gamut of attractive possibilities for modifying inflammation in atherosclerosis, including targeting pivotal inflammatory pathways such as the inflammasomes, inhibiting cytokines, manipulating adaptive immunity and promoting pro-resolution mechanisms. Along with lifestyle measures, pharmacological interventions to mute inflammation could complement traditional targets, such as lipids and hypertension, to make new inroads into the management of atherosclerotic risk.

Targeting Inflammatory Pathways in Cardiovascular Disease: The Inflammasome, Interleukin-1, Interleukin-6 and Beyond.

Recent clinical trials have now firmly established that inflammation participates causally in human atherosclerosis. These observations point the way toward novel treatments that add to established therapies to help stem the growing global epidemic of cardiovascular disease. Fortunately, we now have a number of actionable targets whose clinical exploration will help achieve the goal of optimizing beneficial effects while avoiding undue interference with host defenses or other unwanted actions. This review aims to furnish the foundation for this quest by critical evaluation of the current state of anti-inflammatory interventions within close reach of clinical application, with a primary focus on innate immunity. In particular, this paper highlights the pathway from the inflammasome, through interleukin (IL)-1 to IL-6 supported by a promising body of pre-clinical, clinical, and human genetic data. This paper also considers the use of biomarkers to guide allocation of anti-inflammatory therapies as a step toward realizing the promise of precision medicine. The validation of decades of experimental work and association studies in humans by recent clinical investigations provides a strong impetus for further efforts to target inflammation in atherosclerosis to address the considerable risk that remains despite current therapies.

Heat Shock Protein-90 Inhibition Alters Activation of Pancreatic Stellate Cells and Enhances the Efficacy of PD-1 Blockade in Pancreatic Cancer.

Pancreatic ductal adenocarcinoma (PDAC) has a prominent fibrotic stroma, which is a result of interactions between tumor, immune and pancreatic stellate cells (PSC), or cancer-associated fibroblasts (CAF). Targeting inflammatory pathways present within the stroma may improve access of effector immune cells to PDAC and response to immunotherapy. Heat shock protein-90 (Hsp90) is a chaperone protein and a versatile target in pancreatic cancer. Hsp90 regulates a diverse array of cellular processes of relevance to both the tumor and the immune system. However, to date the role of Hsp90 in PSC/CAF has not been explored in detail. We hypothesized that Hsp90 inhibition would limit inflammatory signals, thereby reprogramming the PDAC tumor microenvironment to enhance sensitivity to PD-1 blockade. Treatment of immortalized and primary patient PSC/CAF with the Hsp90 inhibitor XL888 decreased IL6, a key cytokine that orchestrates immune changes in PDAC at the transcript and protein level in vitro XL888 directly limited PSC/CAF growth and reduced Jak/STAT and MAPK signaling intermediates and alpha-SMA expression as determined via immunoblot. Combined therapy with XL888 and anti-PD-1 was efficacious in C57BL/6 mice bearing syngeneic subcutaneous (Panc02) or orthotopic (KPC-Luc) tumors. Tumors from mice treated with both XL888 and anti-PD-1 had a significantly increased CD8+ and CD4+ T-cell infiltrate and a unique transcriptional profile characterized by upregulation of genes associated with immune response and chemotaxis. These data demonstrate that Hsp90 inhibition directly affects PSC/CAF in vitro and enhances the efficacy of anti-PD-1 blockade in vivo.

Targeting miR-18a sensitizes chondrocytes to anticytokine therapy to prevent osteoarthritis progression

Inflammation participates in the development of OA and targeting inflammatory signaling pathways is a potential strategy for OA treatment. IL-1β is one of the most important inflammatory factors to trigger the activation of NF-κB signaling and accelerate OA progression, whereas OA patients could hardly benefit from inhibiting IL-1β in clinic, suggesting the importance to further explore the details of OA inflammation. We here showed that expression of miR-18a in chondrocytes was specifically induced in response to IL-1β in vitro as well as in rat model of OA during which NF-κB signaling was involved, and that nuclear-translocated p65 directly upregulated miR-18a expression at transcriptional level. Further, increased miR-18a mediated hypertrophy of chondrocytes, resulting in OA degeneration, by targeting TGFβ1, SMAD2, and SMAD3 and subsequently leading to repression of TGF-β signaling. And the level of serum miR-18a was positively correlated to severity of OA. Interestingly, other than IL-1β, pro-inflammation cytokines involving TNFα could also remarkably upregulate miR-18a via activating NF-κB signaling and subsequently induce chondrocytes hypertrophy, suggesting a pivotal central role of miR-18a in inflammatory OA progression. Thus, our study revealed a novel convergence of NF-κB and TGF-β signaling mediated by miR-18a, and a novel mechanism underlying inflammation-regulated OA dependent of NF-κB/miR-18a/TGF-β axis. Notably, in vivo assay showed that targeting miR-18a sensitized OA chondrocytes to IL-1β inhibitor as targeting IL-1β and miR-18a simultaneously had much stronger inhibitory effects on OA progression than suppressing IL-1β alone. Therefore, the diagnostic and therapeutic potentials of miR-18a for OA were also revealed.

Targeting inflammatory pathways by dietary agents for prevention and therapy of cancer

Chronic infections, obesity, alcohol, tobacco, radiation, environmental pollutants, and high-calorie diet have been recognized as major risk factors for the most common types of cancers. All these risk factors are linked to cancer through inflammation. While acute inflammation that persists for short-term mediates host defense against infections, chronic inflammation that lasts for long-term can predispose the host to various chronic illnesses, including cancer. Linkage between cancer and inflammation is indicated by numerous lines of evidence; first, transcription factors NF-κ and STAT3, two major pathways for inflammation, are activated by most cancer risk factors; second, an inflammatory condition precedes most cancers; third, NF-κB and STAT3 are constitutively active in most cancers; fourth, hypoxia and acidic conditions found in solid tumors activate NF-κB; fifth, chemotherapeutic agents and gamma irradiation activate NF-κB and lead to chemo-resistance and radio-resistance; sixth, most gene products linked to inflammation, survival, proliferation, invasion, angiogenesis, and metastasis are regulated by NF-κB and STAT3; seventh, suppression of NF-κB and STAT3 inhibits the proliferation and invasion of tumors; and eighth, most chemo-preventive agents mediate their effects through inhibition of NF-κ and STAT3 activation pathways. Thus suppression of these pro-inflammatory pathways may provide opportunities for both prevention and treatment of cancer. The potential of dietary agents in regulation of these inflammatory cell signaling pathways and their role in prevention and therapy of cancer, is discussed.