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: 10842664
• 负责人: Deepa Rastogi
• 金额: $ 43.96万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-15 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10842664
• 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; 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 现在，非原子或Th2-low哮喘被认为是小儿哮喘的主要亚组。与肥胖有关 哮喘是最常见的小儿非原子性哮喘的形式，与疾病高有关 负担，肺功能较差，缺乏对药物的反应/抵抗力。那迫切需要 研究非原子性哮喘的免疫生物学，以鉴定新的治疗靶标。我们和其他人有 以前报道了肥胖哮喘儿童的外周血中的非原子免疫调查，患有 与肺功能相关的Th1/ Th2比和提高的TNF，IL-6，IFNγ和IP-10 与肥胖有关的哮喘缺乏。使用RNA-seq，我们研究了非原子反应的生物学 刺激肥胖的哮喘CD4+（Th）细胞，发现了几个基因的更新（Dock5，Vav2， CDC42EP4，PAK3，MLK3和PLD1）在细胞分裂周期42（CDC42）途径中。 CDC42EP4和 Dock5基因表达与肥胖哮喘儿童的气道阻塞较差有关。磷酸化 p38，MLK3的下游，并与哮喘中的立体声抗性有关，在刺激的肥胖症中更高 哮喘的TH细胞。小型干扰RNA（siRNA）介导的Cdc42沉默在TH细胞中导致IFNγ和 TNF，但不是IL-4，基因表达。总之，这些结果表明CDC42途径的新作用 肥胖相关哮喘中的非原子炎症。基于这些观察，我们假设 肥胖的哮喘患者，非第2个细胞中Cdc42途径的上调，该途径富含在 气道以及CDC42调节的信号通路的激活，有助于立体声电阻和 伯恩疾病。为了检验我们的假设，我们将使用CDC42途径更新确定非第2个单元 并量化肥胖哮喘患者中Cdc42调节的信号通路的激活。我们将调查 在50种非原子肥胖症中，非第三个细胞在外周血中的富集，而50个 非正常的正常体重哮喘患者，50种肥胖的非哮喘患者和50个健康对照。丰富 气道中的非第三个细胞将在20种肥胖哮喘患者的一部分中进行研究，相比20 正常体重哮喘。在非第2个细胞中缺乏或减毒细胞因子抑制作用 地塞米松将提供证据，表明细胞具有类固醇的耐药性，并在 CDC42和/或CDC42调节的信号通路抑制作用将支持Cdc42在立体声电阻中的作用 在非原子哮喘中。为了确定肥胖的贡献以及肥胖以外的其他因素，我们将比较 非原子肥胖哮喘患者的发现。最后，将CDC42激活与 伯恩（Burnen）疾病，我们将确定对Cdc42激活预测的生化特征，并研究其 小儿非特定肥胖相关哮喘中疾病燃烧的贡献。这些研究将证实 非原子哮喘的免疫生物学和疾病伯嫩中的Cdc42途径，将确定非 - 信号通路中的Th2细胞和/或蛋白作为与肥胖相关哮喘的新型治疗靶标。