Role of the Prolyl Isomerase Pin1 in the Development and Treatment of Asthma

脯氨酰异构酶 Pin1 在哮喘发生和治疗中的作用

• 批准号: 8522222
• 负责人: Kun Ping Lu
• 金额: $ 70.16万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-15 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8522222
• 关键词: Allergens; Animal Model; Asthma; Attenuated; Biological Process; Breathing; Catalysis; Cell model; Cells; Cellular biology; Chronic; Collaborations; Cytokine Activation; Development; Disease; Drug Targeting; Drug usage; Enzymes; Epithelial Cells; Event; Extrinsic asthma; Gene Activation; Genes; Goals; House Dust Mite Allergens; Human; IRAK1 gene; Immune; In Vitro; Incidence; Inflammatory; Interferons; Interleukin-1 Receptors; Israel; Knock-out; Knockout Mice; Lead; Ligation; Link; Lung; Malignant Neoplasms; Mediating; Modality; Modeling; Molecular; Molecular Conformation; Molecular Models; Mus; Natural Immunity; Normal Cell; Outcome; Ovalbumin; Pathway interactions; Peptidylprolyl Isomerase; Pharmaceutical Preparations; Phenotype; Phosphorylation; Physiological; Play; Prevalence; Production; Pulmonary Eosinophilia; Pyroglyphidae; Regulation; Research; Role; Sampling; Signal Pathway; Signal Transduction; Signaling Pathway Gene; Specimen; TLR7 gene; Testing; Th2 Cells; Therapeutic; adaptive immunity; base; cytokine; design; effective therapy; eosinophil; inhibitor/antagonist; mRNA Stability; molecular modeling; mouse model; mutant; new therapeutic target; novel; overexpression; release of sequestered calcium ion into cytoplasm; research study; response; structural biology; therapy development; toll-like receptor 4; transcription factor

DESCRIPTION (provided by applicant): The prevalence of asthma is increasing sharply, but the underlying mechanisms are not fully understood and the therapeutic modalities remain limited. Asthma is a chronic inflammatory disease of the airway induced by overexpression of proinflammatory genes that are regulated by signal pathways in response to various allergens. [For example, it has been shown that house dust mites (HDM) allergens activate TLR4 signaling in epithelial cells to produce cytokines, including IL-33, which activates interleukin-1 receptor 1 (ST2) signaling in Th2 cells and Th2 target cells to induce asthma phenotypes]. A major regulatory mechanism in these signal pathways and gene activation is Pro-directed phosphorylation (pS/T-P). We have previously found that certain pS/T-P motifs can exist in the two completely distinct cis and trans conformations and also identified a unique prolyl isomerase called Pin1 that accelerates their conversion. Importantly, such Pin1-catalyzed conformational regulation often functions as a new molecular timer to act on multiple pathways to coordinate a given biological process to one direction. Significantly, Pin1 is tightly regulated and its deregulation contributes to disease, including asthma, where Pin1 is activated in the airway of human asthma. However, the role of Pin1 in the development and treatment of asthma is not fully understood. Recently, Hypothesizing a role for Pin1 in TLR signaling, we have shown that TLR7/9 ligation activates Pin1, which then activates IRAK1 in TLR7/9 signaling and type I IFN-mediated innate and adaptive immunity. Our preliminary results showed that IL-33 also activated Pin1, whose knockout fully abolished IRAK1 activation, [NF-?B activation], Th2 cytokine production and asthma-like phenotypes induced by IL-33 [or OVA challenge], and potently inhibited cytokine production induced by HDM or LPS in vitro and/or in mice. We identified [several new Pin1 inhibitors, including one that blocked Pin1 activation and cytokine secretion induced by IL-33 in immune cells]. These results led us to hypothesize that Pin1 regulates [TLR4 and ST2 signal pathways] in multiple cells in asthma and is a new drug target for asthma. This proposal is designed to test this novel hypothesis in collaborations with the asthma research expert Elliot Israel and structural expert Linda Nicholson. First, we will use our Pin1 germline and conditional knockout mice to determine the role of Pin1 in regulating [TLR4 and ST2 signaling] in different cells in asthma. Next, we will elucidate the molecular mechanisms of Pin1 function in asthma by determining how Pin1 is activated [by TLR4 and ST2 signaling], how Pin1 regulates IRAK1 and its targets [in TLR4 and ST2 pathways] using cell and mouse asthma models and human asthma specimens, and how Pin1 activates IRAK1 at the structural level. Finally, we will test our new Pin1 inhibitors in cellular and mouse asthma models, and human asthma samples to evaluate their potential as a new therapy for asthma. These studies would uncover a novel mechanism in asthma development, and also could lead to more effective therapies because Pin1 inhibitors, especially when inhaled, might inhibit multiple targets in different cells in asthma.

描述（由申请人提供）：哮喘的患病率急剧增加，但是尚未完全了解潜在的机制，并且治疗方式仍然有限。哮喘是促炎基因过表达引起的气道的慢性炎症性疾病，这些基因受信号途径对各种过敏原的调节。 [例如，已经表明，房屋粉尘螨（HDM）过敏原激活上皮细胞中的TLR4信号传导，以产生包括IL-33在内的细胞因子，它激活了白介素-1受体1 （ST2）TH2细胞和Th2靶细胞中的信号传导诱导哮喘表型]。这些信号途径和基因激活中的主要调节机制是指导磷酸化（PS/T-P）。我们以前已经发现，在两个完全不同的顺式和反式构象中可以存在某些PS/T-P基序，并且还鉴定了一个称为PIN1的独特的prolyl异构酶，该异构酶可以加速其转化率。重要的是，这种PIN1催化的构象调节通常是一种新的分子计时器，以在多种途径上起作用，以将给定的生物学过程与一个方向进行协调。值得注意的是，PIN1受到严格调节，其放松管制导致疾病，包括哮喘，其中PIN1在人类哮喘的气道中被激活。但是，PIN1在哮喘的发育和治疗中的作用尚不完全了解。最近，假设PIN1在TLR信号传导中的作用，我们表明TLR7/9连接激活PIN1，然后激活TLR7/9信号传导中的IRAK1以及I型IFN介导的先天性和适应性免疫。我们的初步结果表明，IL-33还激活了PIN1，其敲除完全废除的IRAK1激活，[NF-？B激活]，Th2细胞因子产生和IL-33诱导的类似哮喘的表型[或OVA挑战]，并有效地抑制了HDM或/或/或/或/或/或/或/或/或/或/或/或/或/或/或/或/或/或/或/或/或/或或或或或或或或或或或或或或或或或或一位为/或/或/或所示。我们鉴定了[几种新的PIN1抑制剂，其中包括阻断IL-33在免疫细胞中诱导的PIN1激活和细胞因子分泌的抑制剂]。这些结果使我们假设PIN1调节哮喘中多个细胞中的PIN1 [TLR4和ST2信号途径]，并且是哮喘的新药物靶标。该建议旨在与哮喘研究专家艾略特（Elliot）和结构性专家琳达·尼科尔森（Linda Nicholson）合作检验这一新假设。首先，我们将使用PIN1种系和条件敲除小鼠来确定PIN1在哮喘中不同细胞中调节[TLR4和ST2信号传导]中的作用。 Next, we will elucidate the molecular mechanisms of Pin1 function in asthma by determining how Pin1 is activated [by TLR4 and ST2 signaling], how Pin1 regulates IRAK1 and its targets [in TLR4 and ST2 pathways] using cell and mouse asthma models and human asthma specimens, and how Pin1 activates IRAK1 at the structural level.最后，我们将在细胞和小鼠哮喘模型以及人哮喘样本中测试我们的新PIN1抑制剂，以评估其作为哮喘的新疗法的潜力。这些研究将发现哮喘发育中的新机制，也可能导致更有效的疗法，因为PIN1抑制剂（尤其是吸入时）可能会抑制哮喘不同细胞中的多个靶标。