We recently found that myometrial distension stimulates maternal uterine vascular remodeling, and hypothesized that this may be a previously unrecognized mechanism for inducing arterial growth during pregnancy. The aim of this study was to further characterize a recently developed surgical model in which medical-grade silicone was infused into one uterine horn of a non-gravid rat to induce acute myometrial stretch, followed by an additional, gradual distension due to the secretion of an exudate into the uterine lumen. Our objectives were to better understand the effects of stretch on the myometrium and to look for the expression of proangiogenic and proinflammatory factors that may stimulate uterine vascular remodeling. Morphometric analysis showed hypertrophy of the uterine corpus that was primarily due to axial growth since the myometrial cross-sectional area was unchanged due to a thinning of the uterine wall secondary to stretch. This finding was supported by significantly increased myometrial smooth muscle cell mitosis. There was also an increase in the concentration of myometrial elastin but not collagen. The analysis showed modest increases in neutrophils, activated and unactivated macrophages, and the proinflammatory cytokines RANTES, MIP-3α, GRO-KC, and TNFα. The most dramatic change was the extremely high level of VEGF in the exudate, which was increased >900× above circulating levels.

Abstract Lunsekimig is a novel, bispecific NANOBODY® molecule that inhibits both thymic stromal lymphopoietin (TSLP) and interleukin (IL)‐13, two key mediators of asthma pathophysiology. In this first‐in‐human study, we evaluated the safety, tolerability, pharmacokinetics (PK), pharmacodynamics (PD), and immunogenicity of lunsekimig in healthy adult participants. Participants received single ascending doses (SAD) of lunsekimig (10–400 mg intravenous [IV] or 400 mg subcutaneous [SC]) (SAD part) or multiple ascending doses (MAD part) of lunsekimig (100 or 200 mg, every 2 weeks [Q2W] for three SC doses), or placebo. Overall, 48 participants were randomized 3:1 in the SAD part and 4:1 in the MAD part for lunsekimig or placebo. The primary endpoint was safety and tolerability. The secondary endpoints included PK, antidrug antibodies (ADAs) and total target measurement. Lunsekimig was well tolerated and common treatment‐emergent adverse events were COVID‐19, nasopharyngitis, injection site reactions, and headache. Lunsekimig showed dose‐proportional increases in exposure and linear elimination. Mean t1/2z of lunsekimig was around 10 days across all IV and SC doses of the SAD and MAD parts of the study. Increases in the serum concentration of total TSLP and IL‐13 for lunsekimig versus placebo indicated target engagement. ADA of low titers were detected in four (11.1%) participants who received lunsekimig in the SAD, and seven (43.8%) in the MAD. In conclusion, lunsekimig was well tolerated in healthy participants with a linear PK profile up to single 400 mg IV and SC dose and multiple doses of 100 and 200 mg SC Q2W, with low immunogenicity.

Introduction: SAR443765 is a novel anti-thymic stromal lymphopoietin (TSLP)/anti-IL-13 engineered antibody (nanobody molecule) shown to significantly and rapidly reduce FeNO in patients with mild-to-moderate asthma after one dose.1 Combined TSLP/IL-13 blockade is expected to improve lung function, particularly small airway dysfunction, which have been shown to correlate with poor disease control and type 2 inflammation. Aims/Objectives: To assess the effects of SAR443765 in asthma. Methods: In a Phase 1 randomized, controlled study (NCT05366764), 36 patients with mild-to-moderate asthma received one subcutaneous SAR443765 400 mg dose. Lung physiology was assessed via spirometry and forced oscillometry technique for up to 57 days. Results: Rapid FEV1 improvement was observed after a single dose of SAR443765 (maximal at Day 8 and largely maintained to Day 57), reflected in reduced respiratory resistance at 5Hz. Positive SAR443765 effects were also observed in forced expiratory flow at 25%–75%, oscillometry resistance heterogeneity (R5-20), and reactance area (AX), particularly in patients with impaired baseline lung function, consistent with improvement in small airway dysfunction. Conclusions: A single dose of the novel anti-TSLP/anti-IL-13 biologic nanobody molecule SAR443765 improved lung function and small airway dysfunction in asthma. These effects surpass those reported for monovalent IL-13 or TSLP pathway targeting and suggest potential for superior effects in patients with asthma. Study and medical writing funded by Sanofi 1Deiteren et al. ATS 2023.

Obesity alters the risks and outcomes of inflammatory lung diseases. It is important to accurately recapitulate the obese state in animal models to understand these effects on the pathogenesis of disease. Diet‐induced obesity is a commonly used model of obesity, but when applied to other disease models like acute respiratory distress syndrome, pneumonia, and asthma, it yields widely divergent. We hypothesized high‐fat chow storage conditions would affect lipid oxidation and inflammatory response in the lungs of lipopolysaccharide (LPS)‐challenged mice. For 6 weeks, C57BL/6crl mice were fed either a 10% (low‐fat diet, LFD) or 60% (high‐fat diet, HFD) stored at room temperature (RT, 23°C) for up to 7, 14, 21, or 42 days. Mice were treated with nebulized LPS to induce lung inflammation, and neutrophil levels in bronchoalveolar lavage were determined 24 h later. Lipid oxidation (malondialdehyde, MDA) was assayed by thiobarbituric acid reactive substances in chow and mouse plasma. Concentrations of MDA in chow and plasma rose in proportion to the duration of RT chow storage. Mice fed a HFD stored <2 weeks at RT had an attenuated response 24 h after LPS compared with mice fed an LFD. This effect was reversed after 2 weeks of chow storage at RT. Chow stored above freezing underwent lipid oxidation associated with significant alterations in the LPS‐induced pulmonary inflammatory response. Our data show that storage conditions affect lipid peroxidation, which in turn affects pulmonary inflammatory responses in a mouse model of disease. It also suggests changes in the microbiome, although not significantly different suggests decreased variety and richness of bacteria in the gut, a large aspect of the immune system. Dietary composition and storage of chow may also affect pulmonary inflammation and the gut microbiome in humans.

Background: Selective inhibition of Receptor interacting serine/threonine protein kinase 1 (RIPK1) could alleviate the hyperinflammatory syndrome occurring in patients with severe coronavirus 2019 (COVID-19). Aim: To explore safety, immunomodulatory and clinical effects of RIPK1 inhibitor, SAR443122 in patients with severe COVID-19. Methods: Patients were randomised (2:1) to receive SAR443122 600mg (300mg twice daily; treatment) or placebo up to 14 days (D; NCT04469621). Safety was assessed up to 28D. The primary outcome was relative change from baseline in C-reactive protein (CRP) level on D7. The time to clinical improvement using 7-point ordinal scale, ventilator/respiratory failure free days (VFD/RFFD), change in SpO2/FiO2 ratio, biomarkers (LDH, IL-6, IL-8) were also explored. Results: The treatment difference in relative change from baseline in CRP is shown in Table. Median time to 50% decrease in CRP level from baseline was 3 vs 5D (p=0.056) in treatment vs placebo groups. Median time for ≥2-point improvement in 7-point scale was 8 vs 10D in treatment vs placebo (p=0.38). Consistent trends towards greater improvements were observed in treatment vs placebo in mean VFD/RFFD, change in SpO2/FiO2 ratio and biomarkers (Table). SAR443122 was considered safe and well tolerated. Conclusion: These preliminary results indicate the need for a larger confirmatory trial to further assess clinically significant effects.

Muscle may contribute to the systemic inflammatory environment during critical illness, but leukocyte interaction and cytokine influence on muscle and its response has not been fully explored in this context. Using an in vivo model of intratracheal lipopolysaccharide (IT LPS)-induced acute lung injury, we show that skeletal muscle rapidly responds with expression of proinflammatory genes, which may be explained by migration of LPS into the circulation. Treatment of mature C2C12 myotubes with LPS at a level achieved in the circulation following IT LPS elicited a proinflammatory cytokine expression profile similar to that of in vivo murine muscle following IT LPS. Stimulation with toll-like receptor (TLR) 2 and 3 agonists provoked comparable responses in C2C12 myotubes. Additionally, co-cultures of C2C12 myotubes and bone marrow-derived macrophages (BMDM) identified the capacity of macrophages to increase myotube proinflammatory gene expression, with tumor necrosis factor-alpha (TNF alpha) gene and protein expression largely attributable to BMDM. To investigate the contribution of TNF alpha in the synergy of the co-culture environment, C2C12 myotubes were treated with recombinant TNF alpha, co-cultures were established using TNF-deficient BMDM, and co-cultures were also depleted of TNF alpha using antibodies. To determine whether the in vitro observations were relevant in vivo, mice received intramuscular administration of LPS +/- TNF alpha or TNF alpha-neutralizing antibodies and showed that TNF alpha is both sufficient and necessary to induce synergistic cytokine release from muscle. Taken together, these data demonstrate how skeletal muscle tissue may contribute proinflammatory cytokines following acute endotoxin injury and the potential of leukocytes to augment this response via TNF alpha secretion.

Background: Pharmacokinetics (PK) of pharmaceuticals and pharmaconutrients are poorly understood in critically ill patients, and dosing is often based on healthy subject data. This might be particularly problematic with enteral medications due to metabolic abnormalities and impaired gastrointestinal tract absorption common in critically ill patients. Utilizing enteral fish oil, this study was undertaken to better understand and define PK of enteral omega-3 fatty acids (eicospentaenoic acid [EPA] and docosahexaenoic acid [DHA]) in critically ill patients with severe sepsis. Materials and methods: Healthy volunteers (n = 15) and mechanically ventilated (MV) adults with severe sepsis (n = 10) were recruited and received 9.75 g EPA and 6.75 g DHA daily in two divided enteral doses of fish oil for 7 days. Volunteers continued their normal diet without other sources of fish oil, and sepsis patients received standard enteral feeding. Blood was collected at frequent intervals during the 14-day study period. Peripheral blood mononuclear cells (PMBCs) and neutrophils were isolated and analyzed for membrane fatty acid (FA) content. Mixed linear models and t-tests were used to analyze changes in FA levels over time and FA levels at individual time points, respectively. PK parameters were obtained based on single compartment models of EPA and DHA kinetics. Results: Healthy volunteers were 41.1 +/- 10.3 years; 67% were women. In patients with severe sepsis (55.6 +/- 13.4 years, 50% women), acute physiologic and chronic health evaluation (APACHE) II score was 27.2 +/- 8.8 at ICU admission and median MV duration was 10.5 days. Serum EPA and DHA were significantly lower in sepsis vs. healthy subjects over time. PBMC EPA concentrations were generally not different between groups over time, while PBMC DHA was higher in sepsis patients. Neutrophil EPA and DHA concentrations were similar between groups. The half-life of EPA in serum and neutrophils was significantly shorter in sepsis patients, whereas other half-life parameters did not vary significantly between healthy volunteers and sepsis patients. Conclusions: While incorporation of n-3 FAs into PBMC and neutrophil membranes was relatively similar between healthy volunteers and sepsis patients receiving identical high doses of fish oil for one week, serum EPA and DHA were significantly lower in sepsis patients. These findings imply that serum concentrations and EPA and DHA may not be the dominant driver of leukocyte membrane incorporation of EPA and DHA. Furthermore, lower serum EPA and DHA concentrations suggest that either these n-3 FAs were being metabolized rapidly in sepsis patients or that absorption of enteral medications and pharmaconutrients, including fish oil, may be impaired in sepsis patients. If enteral absorption is impaired, doses of enteral medications administered to critically ill patients may be suboptimal. (C) 2019 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism. All rights reserved.

Once you are diagnosed with asthma, it is important to work closely with your healthcare provider to control your asthma. This fact sheet describes commonly used asthma medicines and other actions you can take to control asthma.

RationaleIntensive care unit acquired weakness (ICUAW) occurs in nearly 50% of patients admitted to the ICU. Patients exhibiting ICUAW have increased in‐hospital and post‐ICU mortality. ICUAW, and the associated muscle atrophy specifically, cannot be completely explained by disuse and may involve a secondary trigger to enhance the breakdown of muscle fibers. Recent work has suggested that excessive systemic inflammation may contribute to this process. We hypothesized that muscle atrophy in ICUAW may, in part, be induced by myeloid cell recruitment to muscle.MethodsC57Bl6/J mice received intratracheal (IT) instillation of lipopolysaccharide (LPS, 3 μg/g) or saline and were euthanized 3, 6, 12, 24, 72 hours thereafter. For antibody‐mediated myeloid cell depletion, 200 μg of αGR‐1, αLy6G, or isotype control antibodies were administered IP 24 hours and 1 hour prior to injury and mice were examined 24 hours post‐injury. Whole gastrocnemius muscle was analyzed using RT‐qPCR. For in vitro studies, C2C12 myoblasts were differentiated for 7 days and stimulated with LPS (1 μg/ml) to elicit an inflammatory response. Bone marrow derived macrophages (BMDMs) were differentiated with 10 ng/ml of macrophage‐colony stimulation factor for 7 days and seeded at a density of 2600 cells/cm2 (4940 cells/well) onto C2C12 myotubes after 6 days of differentiation. On day 7 of C2C12 differentiation, cocultures were stimulated as previously described. Supernatants and cell lysates were analyzed by ELISA and RT‐qPCR. Myotube diameter was measured using ImageJ.ResultsIT LPS upregulates the atrogene muscle RING‐finger 1 (MuRF1) and induces atrophy in vivo. However, this effect of LPS is not seen in cultured C2C12 myotubes alone. Depletion of neutrophils and monocytes with αGR‐1 antibody reduced MuRF1 expression in vivo. Both C2C12 myotubes and whole muscle show rapid, transient expression of myeloid chemoattractants MCP‐1 and KC (CXCL1). Cocultures of BMDMs and C2C12 myotubes enhance the expression of IL‐6 and MCP‐1 synergistically and induce C2C12 myotubes to produce TNFα.ConclusionsOur data indicate that IT LPS induces MuRF1 in vivo and our in vitro data in myotubes suggests that this effect may require myeloid cells. Further supporting this conclusion, we found that antibody depletion of myeloid cells reduced IT LPS‐induced MuRF1 expression. Moreover, expression of key myeloid cell chemoattractants are induced in muscle both in vitro and in vivo in response to LPS and addition of BMDMs to C2C12 myotubes with LPS treatment reveals synergistic increases of IL‐6 and MCP‐1 and induces C2C12 myotubes to produce TNFα. The current study implicates macrophages and neutrophils as modulators of the muscle atrophy response to following lung injury.Support or Funding InformationNIH 5R01HL143452‐02NIH 2T32HL076122‐16

Protein disulfide isomerases (PDI) are a family of redox chaperones that catalyze formation or isomerization of disulfide bonds in proteins. Previous studies have shown that one member, PDIA3, interacts with influenza A virus (IAV) hemagglutinin (HA), and this interaction is required for efficient oxidative folding of HA in vitro. However, it is unknown whether these host-viral protein interactions occur during active infection and whether such interactions represent a putative target for the treatment of influenza infection. Here we show that PDIA3 is specifically upregulated in IAV-infected mouse or human lung epithelial cells and PDIA3 directly interacts with IAV-HA. Treatment with a PDI inhibitor, LOC14 inhibited PDIA3 activity in lung epithelial cells, decreased intramolecular disulfide bonds and subsequent oligomerization (maturation) of HA in both H1N1 (A/PR8/34) and H3N2 (X31, A/Aichi/68) infected lung epithelial cells. These reduced disulfide bond formation significantly decreased viral burden, and also pro-inflammatory responses from lung epithelial cells. Lung epithelial-specific deletion of PDIA3 in mice resulted in a significant decrease in viral burden and lung inflammatory-immune markers upon IAV infection, as well as significantly improved airway mechanics. Taken together, these results indicate that PDIA3 is required for effective influenza pathogenesis in vivo, and pharmacological inhibition of PDIs represents a promising new anti-influenza therapeutic strategy during pandemic and severe influenza seasons.

Purpose: Bronchoalveolar fluid (BALF) and plasma biomarkers are often endpoints in early phase randomized trials (RCTs) in acute respiratory distress syndrome (ARDS). With ARDS mortality decreasing, we analyzed baseline biomarkers in samples from contemporary ARDS patients participating in a prior RCT and compared these to historical controls.Materials and methods: Ninety ARDS adult patients enrolled in the parent trial. BALF and blood were collected at baseline, day 41, and day 81. Interleukins-8/-6/-1/-1 receptor antagonist/-10; granulocyte colony stimulating factor; monocyte chemotactic protein-1; tumour necrosis factor-; surfactant protein-D; von Willebrand factor; leukotriene B-4; receptor for advanced glycosylation end products; soluble Fas ligand; and neutrophil counts were measured.Results: Compared to historical measurements, our values were generally substantially lower, despite our participants being similar to historical controls. For example, our BALF IL-8 and plasma IL-6 were notably lower than in a 1999 RCT of low tidal volume ventilation and a 2007 biomarker study, respectively.Conclusions: Baseline biomarker levels in current ARDS patients are substantially lower than 6-20years before collection of these samples. These findings, whether from ICU care changes resulting in less inflammation or from variation in assay techniques over time, have important implications for design of future RCTs with biomarkers as endpoints.