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(申请人提供)：哮喘的患病率正在急剧增加，但其潜在的机制尚不完全清楚，治疗方式仍然有限。哮喘是一种慢性呼吸道炎症性疾病，由致炎基因的过度表达引起，这些致炎基因由信号通路调节，以应对各种过敏原。[例如，已经证明，屋尘螨(HDM)变应原激活上皮细胞中的TLR4信号以产生细胞因子，包括IL-33，它激活白细胞介素1受体1 (ST2)Th2细胞和Th2靶细胞中的信号诱导哮喘表型]。这些信号通路和基因激活的主要调控机制是Pro-定向磷酸化(PS/T-P)。我们以前已经发现，某些PS/T-P基序可以存在于两种完全不同的顺式和反式构象中，并发现了一种独特的脯氨基异构酶Pin1，它可以加速它们的转化。重要的是，这种Pin1催化的构象调节通常作为一个新的分子计时器作用于多个途径，以协调一个给定的生物过程朝着一个方向进行。值得注意的是，Pin1受到严格的调控，它的解除调控导致了包括哮喘在内的疾病，其中Pin1在人类哮喘的呼吸道中被激活。然而，Pin1在哮喘的发生和治疗中的作用尚不完全清楚。最近，假设Pin1在TLR信号转导中的作用，我们发现TLR7/9连接激活Pin1，继而激活TLR7/9信号转导中的IRAK1和I型干扰素介导的先天免疫和获得性免疫。我们的初步结果表明，IL-33还激活了Pin1，而Pin1的基因敲除完全阻断了IL-33[或OVA攻击]诱导的IRAK1激活、[NF-B激活]、Th2细胞因子的产生和哮喘样表型，并在体外和/或在小鼠体内有效地抑制了HDM或LPS诱导的细胞因子的产生。我们确定了[几种新的Pin1抑制剂，包括一种可以阻断免疫细胞中IL-33诱导的Pin1激活和细胞因子分泌的药物]。这些结果使我们假设Pin1在哮喘的多个细胞中调节[TLR4和ST2信号通路]，是治疗哮喘的新药物靶点。这项建议旨在与哮喘研究专家埃利奥特·伊斯雷尔和结构专家琳达·尼科尔森合作，测试这一新的假设。首先，我们将使用我们的Pin1胚系和条件基因敲除小鼠来确定Pin1在哮喘不同细胞中调节[TLR4和ST2信号]的作用。接下来，我们将通过确定Pin1是如何被[TLR4和ST2信号]激活的，Pin1如何通过使用细胞和小鼠哮喘模型和人类哮喘标本来调节IRAK1及其靶点[在TLR4和ST2通路中]，以及Pin1如何在结构水平上激活IRAK1来阐明Pin1在哮喘中的分子机制。最后，我们将在细胞和小鼠哮喘模型以及人类哮喘样本中测试我们的新Pin1抑制剂，以评估它们作为哮喘新疗法的潜力。这些研究将揭示哮喘发生的新机制，并可能导致更有效的治疗，因为Pin1抑制剂，特别是当吸入时，可能会在哮喘的不同细胞中抑制多个靶点。