Investigating the CDC42 pathway as a novel pathway for pediatric non-atopic obesity-related asthma

研究 CDC42 通路作为儿童非特应性肥胖相关哮喘的新通路

• 批准号: 10554285
• 负责人: Deepa Rastogi
• 金额: --
• 依托单位: CHILDREN'S RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-01 至 2023-07-21
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10554285
• 关键词: Affect; Albuterol; Asthma; Attenuated; Awareness; Biochemical; Biology; Bronchoalveolar Lavage; CD4 Positive T Lymphocytes; CXCL10 gene; CXCR3 gene; Cell Cycle; Cell physiology; Cells; Child; Childhood; Childhood Asthma; Chronic Disease; Clinical; Cytokine Suppression; Data; Dexamethasone; Drug Targeting; Extrinsic asthma; Flow Cytometry; Gene Expression; Genes; Genotype; Helper-Inducer T-Lymphocyte; Heterogeneity; Immune; Immune response; Immunobiology; In Vitro; Inflammation; Inhalation; Interferon Type II; Interleukin-6; Light; Link; MAPK8 gene; Measures; Mediating; Memory; Metabolic; Monomeric GTP-Binding Proteins; Obesity; Pathway interactions; Pharmaceutical Preparations; Phenotype; Phosphorylation; Play; Proteins; Reporting; Research; Resistance; Risk; Role; Screening procedure; Signal Pathway; Small Interfering RNA; Steroid Resistance; Steroids; Subgroup; T-Lymphocyte; TNF gene; Testing; Th2 Cells; Transcript; Treatment Failure; Up-Regulation; VAV2 gene; Weight; airway obstruction; asthmatic; asthmatic airway; burden of illness; cell type; cohort; cytokine; drug discovery; insight; mTOR Signaling Pathway; new therapeutic target; novel; obesity in children; obesity treatment; obesity-associated asthma; p38 Mitogen Activated Protein Kinase; peripheral blood; predictive signature; promoter; public health relevance; pulmonary function; response; single-cell RNA sequencing; targeted treatment; transcriptome sequencing

ABSTRACT Non-atopic or Th2-low asthma is now recognized as a major subgroup of pediatric asthma. Obesity-related asthma, the most commonly reported form of pediatric non-atopic asthma, is associated with high disease burden, worse lung function, and lack of response/resistance to medications. Thus, there is an urgent need to investigate the immunobiology of non-atopic asthma to identify novel therapeutic targets. We and others have previously reported non-atopic immune responses in peripheral blood from obese asthmatic children, with elevated TH1/ TH2 ratio and increased TNF, IL-6, IFNγ, and IP-10 that correlated with pulmonary function deficits in obesity-related asthma. Using RNA-Seq, we probed the biology of non-atopic responses in un- stimulated obese asthmatic CD4+ (TH) cells and found upregulation of several genes (DOCK5, VAV2, CDC42EP4, PAK3, MLK3 and PLD1) in the Cell Division Cycle 42 (CDC42) pathway. Higher CDC42EP4 and DOCK5 gene expression correlated with worse airway obstruction in obese asthmatic children. Phosphorylated p38, downstream of MLK3, and linked with steroid resistance in asthma, was higher in stimulated obese asthmatic TH cells. Small interfering RNA (siRNA)-mediated CDC42 silencing in TH cells led to lower IFNγ and TNF, but not IL-4, gene expression. Together, these results suggest a novel role for the CDC42 pathway in non-atopic inflammation in obesity-related asthma. Based on these observations, we hypothesize that in obese asthmatics, upregulation of the CDC42 pathway in a non-TH2 TH cell, which is enriched in the airway, and activation of CDC42-regulated signaling pathways, contribute to steroid resistance and disease burden. To test our hypothesis, we will identify the non-TH2 TH cell with CDC42 pathway upregulation and quantify activation of CDC42-regulated signaling pathways in obese asthmatics. We will investigate enrichment of the non-TH2 TH cell in peripheral blood in 50 non-atopic obese asthmatics as compared to 50 non-atopic normal-weight asthmatics, 50 obese non-asthmatics, and 50 healthy controls. Enrichment of the non-TH2 TH cell in the airway will be investigated in a subset of 20 obese asthmatics and compared to 20 normal-weight asthmatics. Absent or attenuated cytokine suppression in the non-TH2 TH cell in response to dexamethasone will provide evidence that the cell is steroid resistant and gain of steroid sensitivity following CDC42 and/or CDC42-regulated signaling pathway inhibition will support a role of CDC42 in steroid resistance in non-atopic asthma. To identify the contribution of obesity, and of factors other than obesity, we will compare the findings in non-atopic obese asthmatics to obese non-asthmatics. Lastly, to link CDC42 activation with disease burden, we will identify a biochemical signature predictive of CDC42 activation, and investigate its contribution to disease burden in pediatric non-atopic obesity-related asthma. These studies will confirm a role of CDC42 pathway in the immunobiology and disease burden of non-atopic asthma, and will identify the non- TH2 TH cell and/or proteins in the signaling pathways as novel therapeutic targets for obesity-related asthma.

抽象的 非特应性或 Th2 低哮喘现在被认为是儿童肥胖相关哮喘的一个主要亚组。 哮喘是最常见的儿童非特应性哮喘形式，与高发病率相关 负担、肺功能恶化以及对药物缺乏反应/耐药性，因此迫切需要治疗。 我们和其他人研究了非特应性哮喘的免疫生物学以确定新的治疗靶点。 先前报道肥胖哮喘儿童外周血中的非特应性免疫反应 TH1/TH2 比率升高，TNF、IL-6、IFNγ 和 IP-10 增加，与肺功能相关 使用RNA-Seq，我们探讨了肥胖相关哮喘的非特应性反应的生物学。 刺激肥胖哮喘 CD4+ (TH) 细胞并发现多个基因（DOCK5、VAV2、 细胞分裂周期 42 (CDC42) 途径中的 CDC42EP4、PAK3、MLK3 和 PLD1) 和高级 CDC42EP4。 DOCK5 基因表达与肥胖哮喘儿童更严重的气道阻塞相关。 p38 是 MLK3 的下游，与哮喘的类固醇抵抗有关，在刺激性肥胖中含量较高 哮喘 TH 细胞中小干扰 RNA (siRNA) 介导的 CDC42 沉默导致 IFNγ 和 TNF 但不是 IL-4 基因表达，这些结果表明 CDC42 通路在基因表达中具有新作用。 基于这些观察，我们将其纳入肥胖相关哮喘的非特应性炎症中。 肥胖哮喘患者，非 TH2 TH 细胞中 CDC42 通路上调，该细胞富含 气道和 CDC42 调节的信号通路的激活有助于类固醇抵抗和 为了检验我们的假设，我们将鉴定具有 CDC42 通路上调的非 TH2 TH 细胞。 我们将研究肥胖哮喘患者中 CDC42 调节的信号通路的激活和量化。 与 50 名非特应性肥胖哮喘患者相比，50 名非特应性肥胖哮喘患者外周血中非 TH2 TH 细胞富集 非特应性正常体重哮喘患者、50 名肥胖非哮喘患者和 50 名健康对照。 气道中的非 TH2 TH 细胞将在 20 名肥胖哮喘患者中进行研究，并与 20 名肥胖哮喘患者进行比较 正常体重的哮喘患者对非 TH2 TH 细胞的细胞因子抑制不存在或减弱。 地塞米松将提供证据证明细胞对类固醇具有抗性并且在以下情况下获得类固醇敏感性 CDC42和/或CDC42调节的信号通路抑制将支持CDC42在类固醇抵抗中的作用 为了确定肥胖和肥胖以外因素的影响，我们将进行比较。 最后，将 CDC42 激活与非特应性肥胖哮喘患者的研究结果联系起来。 疾病负担，我们将确定预测 CDC42 激活的生化特征，并研究其 这些研究将证实其对儿童非特应性肥胖相关哮喘的疾病负担的作用。 CDC42 通路在非特应性哮喘的免疫生物学和疾病负担中的作用，并将识别非特应性哮喘 TH2 信号通路中的 TH 细胞和/或蛋白质作为肥胖相关哮喘的新治疗靶点。