Objectives: To assess the efficacy and safety of hydroxychloroquine (HCQ) plus standard–of–care (SOC) compared with SOC alone in adult patients with COVID–19. Design: Multicenter, open–label, randomized controlled trial. Setting: 16 government–designated COVID–19 treatment centers in China through 11 to 29 in February 2020. Participants: 150 patients hospitalized with laboratory confirmed COVID–19 were included in the intention to treat analysis. 75 patients were assigned to HCQ plus SOC and 75 to SOC alone. Interventions: HCQ was administrated with a loading dose of 1, 200 mg daily for three days followed by a maintained dose of 800 mg daily for the remaining days (total treatment duration: 2 or 3 weeks for mild/moderate or severe patients, respectively). Main outcome measures: The primary outcome was whether participants had a negative conversion of SARS–CoV–2 by 28 days, and was analyzed according to the intention–to–treat principle. Adverse events were analyzed in the safety population in which HCQ recipients were participants who actually received at least one dose of HCQ and HCQ non–recipients were those actually managed with SOC alone. Results: Among 150 patients, 148 were with mild to moderate disease and 2 were with severe disease. The mean days (± standard deviation, min to max) from symptoms onset to randomization was 16.6 (±10.5 days, 3 to 41 days). The negative conversion probability by 28 days in SOC plus HCQ group was 85.4% (95% confidence interval (CI) 73.8% to 93.8%), similar to that in the SOC group 81.3% (95%CI 71.2% to 89.6%). Between–group difference was 4.1% (95%CI –10.3% to 18.5%). In the safety population, adverse events were recorded in 7 (8.8%) HCQ non–recipients (N=80) and in 21 (30%) HCQ recipients (N=70). The most common adverse event in the HCQ recipients was diarrhea, reported in 7 (10%) patients. Two HCQ recipients reported serious adverse events. Conclusions: The administration of HCQ did not result in a significantly higher negative conversion probability than SOC alone in patients mainly hospitalized with persistent mild to moderate COVID–19. Adverse events were higher in HCQ recipients than in HCQ non–recipients. Trial registration: ChiCTR2000029868 What is already known on this topic: — The pandemic of coronavirus disease 2019 (COVID–19) imposes substantial burdens on individuals, communities, health–care facilities, markets, governments, etc. globally. — There is no specific treatment approved for COVID–19 or vaccine to prevent infection with the novel coronavirus. — During the urgent pandemic, media headlines the utility of drugs without solid evidence but buries the side–effects of these drugs. What this study adds: — In this randomized clinical trial of patients mainly with persistent mild to moderate COVID–19, exposure to hydroxychloroquine led to a similar probability of virus elimination comparing to the current standard–of–care. — Adverse events, mostly gastrointestinal related, were significantly increased in patients who received hydroxychloroquine. — Overall, the results from our trial do not support the use of hydroxychloroquine in patients with persistent mild to moderate COVID–19. Print Study question: To assess the efficacy and safety of hydroxychloroquine (HCQ) plus standard–of–care (SOC) compared with SOC alone in adult patients with COVID–19. Methods: This is a multicenter, open–label, randomized controlled trial conducted in 16 government–designated COVID–19 treatment centers in China through 11 to 29 in February 2020. A total of 150 patients hospitalized with laboratory confirmed COVID–19 were included in the intention to treat analysis. Among them, 75 patients were assigned to HCQ plus SOC and 75 to SOC alone. HCQ was administrated with a loading dose of 1, 200 mg daily for three days followed by a maintained dose of 800 mg daily for the remaining days (total treatment duration: 2 or 3 weeks for mild/moderate or severe patients, respectively). The primary outcome was whether participants had a negative conversion of SARS–CoV–2 by 28 days, and was analyzed according to the intention to treat principle. Adverse events were analyzed in the safety population in which HCQ recipients were participants who actually received at least one dose of HCQ and HCQ non–recipients were those actually managed with SOC alone. Study answer and limitations: Among 150 patients, 148 were with mild to moderate disease and 2 were with severe disease. The mean days (± standard deviation, min to max) from symptoms onset to randomization was 16.6 (±10.5 days, 3 to 41 days). The negative conversion probability by 28 days in SOC plus HCQ group was 85.4% (95% confidence interval (CI) 73.8% to 93.8%), similar to that in the SOC group 81.3% (95%CI 71.2% to 89.6%). Between–group difference was 4.1% (95%CI –10.3% to 18.5%). In the safety population, adverse events were recorded in 7 (8.8%) HCQ non–recipients (N=80) and in 21 (30%) HCQ recipients (N=70) with two serious adverse events. The most common adverse event in the HCQ recipients was diarrhea, reported in 7 (10%) patients. Two HCQ recipients reported serious adverse events. What this study adds: Our trial does not support the use of hydroxychloroquine in patients with persistent mild to moderate COVID–19 due to limited effects on virus eliminating and significantly increased adverse events. Funding, competing interests, data sharing: This work was supported by the Emergent Projects of National Science and Technology (2020YFC0844500), National Natural Science Foundation of China (81970020, 81770025), National Key Research and Development Program of China (2016YFC0901104), Shanghai Municipal Key Clinical Specialty...

• miR-210 level is essential for cell-cycle progression in cortical neural progenitors • Cdk7 and miR-210 control neural progenitor proliferation • miR-210 promotes premature cell-cycle exit and differentiation in neural progenitors • miR-210 expression induces a deep-layer neuronal fate in the neocortex

Chen H, et al. Allergy Asthma Immunol Res. 2021 Mar;13(2):177-205. https://doi.org/10.4168/aair.2021.13.2.177

Background: Eosinophils act as a secondary antigen-presenting cell (APC) to stimulate Th cell responses against antigens. IL-25 plays a significant role in eosinophil activation in allergic asthma. The role of IL-25 on the classic APC functions of dendritic cells has been elucidated. However, whether IL-25 facilitates eosinophils for antigen presentation is unknown.Objective: To elucidate the role of IL-25 on eosinophils antigen presenting function during allergic asthma and its related mechanism.Methods: Eosinophils from allergic asthma subjects were cultured with IL-25 and HDM to identify the co-stimulator molecules expression. Co-cultures of patient eosinophils and autologous naïve CD4⁺ T cells in the same culture system were to explore whether eosinophils had the capacity to promote Th cell differentiation in response to IL-25 engagement. In asthma mouse model, IL-25^-/- mice were exposed to HDM to investigate the effect of IL-25 on eosinophils during the sensitization phase. The impact of IL-25 on the capacity for eosinophils taking up antigens was evaluated. Mouse bone marrow derived eosinophils (BmEOS) were co-cultured with naïve CD4⁺T cells sorted from spleens under HDM and IL-25 stimulation to identify T cell differentiation.Results: IL-25 upregulated HLA-DR, PD-L1, and OX-40L expression on eosinophils from allergic asthma patients. IL-25 and HDM co-sensitized eosinophils promoted Th2 differentiation. In mouse model, IL-25^-/- mice experienced restrained allergic pulmonary inflammation and reduced eosinophils recruitment and antigen uptake capacity during the early sensitization phase. In vitro, IL-25 promoted antigen uptake by eosinophils. In BmEOS and naïve CD4⁺T cells co-culture, IL-25 accreted the proportion of CD4⁺Th2 cells, which was absent in CD4⁺T cells culture alone.Conclusion: Our data identify a novel role of IL-25 in enhancing eosinophils antigen uptake and co-stimulator molecules expression to induce Th2 priming in the context of allergic inflammation.

N6-methyladenosine (m6A) modification has been implicated in many cell processes and diseases. YTHDF1, a translation-facilitating m6A reader, has not been previously shown to be related to allergic airway inflammation. Here, we report that YTHDF1 is highly expressed in allergic airway epithelial cells and asthmatic patients and that it influences proinflammatory responses. CLOCK, a subunit of the circadian clock pathway, is the direct target of YTHDF1. YTHDF1 augments CLOCK translation in an m6A-dependent manner. Allergens enhance the liquid-liquid phase separation (LLPS) of YTHDF1 and drive the formation of a complex comprising dimeric YTHDF1 and CLOCK mRNA, which is distributed to stress granules. Moreover, YTHDF1 strongly activates NLRP3 inflammasome production and interleukin-1β secretion leading to airway inflammatory responses, but these phenotypes are abolished by deleting CLOCK. These findings demonstrate that YTHDF1 is an important regulator of asthmatic airway inflammation, suggesting a potential therapeutic target for allergic airway inflammation.

Eosinophil-lineage committed progenitor cells (EoP) migrate from the bone marrow and differentiate locally to provide an ongoing source of mature eosinophils in asthmatic inflammatory responses in the airways. Sputum levels of EoP are increased in asthmatics compared to normal controls suggesting an exaggerated eosinophilopoietic environment in the airways. Understanding what factors promote EoP traffic to the airways is important to understand the diathesis of asthma pathology. Interleukin (IL)-25, is an epithelial-derived cytokine that promotes type 2 inflammatory responses. We have previously shown that levels of IL-25 and expression of the IL-25 receptor (IL-17RA and IL-17RB) on mature eosinophils are greater in allergic asthmatics compared to atopic non-asthmatics and non-atopic normal controls. In addition, these levels were increased significantly increased following allergen inhalation challenge and physiologically relevant levels of IL-25 stimulated eosinophil degranulation, intracellular IL-5 and IL-13 expression and primed migration to eotaxin. The current study, examined the role of IL-25 on allergen-induced trafficking of EoP in atopic asthmatics. Asthmatics (n = 14) who developed allergen-induced early and late responses were enrolled in the study. Blood was collected at pre- and 24 h post-challenge. At each time point, surface expression of IL-17RA and IL-17RB on EoP was evaluated by flow cytometry. Migration assays examined the effect of IL-25 on EoP chemotactic responses, in vitro. In addition, IL-25 knockout ovalbumin (OVA) sensitized and challenged mice were studied to evaluate in vivo mobilization effects of IL-25 on newly formed EoP and mature eosinophils. There was a significant increase in numbers of blood EoP expressing IL-17RB, 24 h post-allergen inhalation challenge in allergic asthmatics. Pre-exposure to IL-25 primed the migrational responsiveness of EoP to stromal cell-derived factor 1α. In OVA-sensitized mice, knocking out IL-25 significantly alleviated OVA-induced eosinophil infiltration in the airway and newly formed eosinophils were reduced in the lung. The findings of this study indicate a potential role for IL-25 in allergen-induced trafficking of EoP to the airways and local differentiation promoting tissue eosiniophilia in asthmatic responses.