Taming IL-33 to Control Inflammation and Fibrosis

驯服 IL-33 来控制炎症和纤维化

基本信息

批准号：
10615887
负责人：
Irina G. Luzina
金额：
$ 33.99万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-06-01 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10615887
关键词：
Adaptor Protein Complex 2 Adipose tissue Attention Automobile Driving Binding Biology Cell Nucleus Cell Surface Receptors Cells Chromatin Cicatrix Complex Cytoplasm Data Development Disease Fibrosis Future Goals Heart Importins Inflammation Inflammatory Inflammatory Response Innovative Therapy Interleukins Kidney Knockout Mice Length Ligands Liver Lung Maps Mediating Medicine Modality Molecular N-terminal Nuclear Organ Pathology Patients Peptides Peritoneum Permeability Phenotype Phosphorylation Process Proteins Regulation Reporting Research Role Signal Transduction Skeletal Muscle Skin Solid Source Stress Therapeutic Tissues Transcriptional Regulation Transforming Growth Factor beta Work attenuation cell type cytokine exhaust extracellular in vivo inflammatory modulation innovation novel prospective receptor restraint sensor wound healing

项目摘要

Inflammation and fibrosis often co-occur and contribute to a remarkably broad variety of diseases of every organ and tissue. The overall role of interleukin (IL)-33 in inflammation and fibrosis has already been established, but the mechanisms of such regulation are not fully understood. Limited attention has been paid to the IL-33 precursor—full-length IL-33 (FLIL33)—which is basally and inducibly expressed, resides mostly in the cell nucleus, and is thought to regulate inflammatory responses, wound healing, and transcriptional regulation independently of the mature IL-33 (MIL33) cytokine. The understudied FLIL33 requires more attention, because it is both the immediate source of the MIL33 cytokine and an independently active factor. These IL-33 forms become pathophysiologically engaged under stress, but IL-33-null mice have no noticeable basal phenotype, suggesting that IL-33 depletion is a safe therapeutic approach. Our objective is to form the basis for therapeutic manipulation of IL-33 in inflammatory fibrotic diseases through integrated understanding of proteolytic maturation and extracellular release of MIL33, intracellular signaling and functioning of FLIL33, and proteolytic stability of the FLIL33 protein pool. We amassed new data related to the molecular control of IL-33 subcellular localization, functional maturation, and extracellular release through a previously unknown region within the FLIL33 molecule, which spans substantially more of the N-terminus than the currently known “sensor domain.” We hypothesize that this segment may be targeted to control IL-33 activation and extracellular release. We also discovered that the intracellular function of FLIL33 is centered on Smad3 phosphorylation in a TGF-beta-independent fashion. We hypothesize that this process is mediated by the adaptor-related protein complex 2 and that targeting this mechanism allows for abrogation of the functional effects of intracellular FLIL33. We also recently reported that importin 5 (IPO5) protects FLIL33 from proteolytic degradation, driving the hypothesis that IPO5-binding, cell-permeable decoy peptide(s) will induce the loss of IPO5-mediated protection of FLIL33 from proteasomal degradation. This prospective therapy will deplete the FLIL33 protein pool, thus exhausting the source of MIL33 and simultaneously attenuating the intracellular effects ofFLIL33. The Specific Aims of this project are to: 1) Precisely map the segments in the N-terminal region of FLIL33 that are responsible for its nuclear-versus-cytoplasmic localization, functional maturation, extracellular release, and selective binding of intracellular molecular partners; 2) Define the molecular mechanism responsible for FLIL33-induced, TGF-βligand-independent phosphorylation of Smad3; and 3) Develop a cell-permeable decoy peptide-based approach to deplete intracellular FLIL33, thereby exhausting the source of MIL33 and simultaneously attenuating the independent effects of FLIL33. Evaluate the in vivo efficacy of such FLIL33 depletion. Achieving these goals will offer new understanding of IL-33 pathobiology and initiate the development of precise, innovative therapeutic modalities.

炎症和纤维化经常同时发生，并导致各种不同寻常的疾病。白细胞介素 (IL)-33 在炎症和纤维化中的总体作用已被证实。已建立，但这种监管机制尚未得到充分重视。 IL-33 前体——全长 IL-33 (FLIL33)——基础性和诱导性表达，主要存在于存在于细胞核中，被认为可以调节炎症反应、伤口愈合和转录成熟 IL-33 (MIL33) 细胞因子的独立调节正在研究的 FLIL33 需要更多。引起关注，因为它既是 MIL33 细胞因子的直接来源，又是一个独立的活性因子。这些 IL-33 形式在压力下会发生病理生理学变化，但 IL-33 缺失小鼠没有明显的变化基础表型，表明 IL-33 耗竭是一种安全的治疗方法。通过综合理解 IL-33 在炎症纤维化疾病中的治疗操作奠定基础 MIL33 的蛋白水解成熟和细胞外释放、FLIL33 的细胞内信号传导和功能，我们收集了与分子控制相关的新数据。 IL-33 亚细胞定位、功能成熟和通过先前未知的细胞外释放 FLIL33 分子内的区域，其 N 末端的范围比目前已知的要大得多 “传感器结构域。”我们发现该片段可能旨在控制 IL-33 的激活和我们还发现FLIL33的细胞内功能以Smad3为中心。我们发现这一过程是由 TGF-β 介导的。接头相关蛋白复合物 2 以及靶向该机制可以废除功能性我们最近还报道了 importin 5 (IPO5) 可以保护 FLIL33 免受细胞内 FLIL33 的影响。蛋白水解降解，推动了 IPO5 结合、细胞可渗透的诱饵肽将诱导的假设这种前瞻性疗法将导致 FLIL33 失去 IPO5 介导的保护，免受蛋白酶体降解。耗尽 FLIL33 蛋白库，从而耗尽 MIL33 的来源，同时减弱 FLIL33 的细胞内效应该项目的具体目标是： 1) 精确绘制 FLIL33 中的片段。 FLIL33 的 N 末端区域负责其核与细胞质定位、功能细胞内分子伴侣的成熟、细胞外释放和选择性结合；2) 定义 FLIL33 诱导的、不依赖于 TGF-β 配体的 Smad3 磷酸化的分子机制； 3) 开发一种基于细胞渗透性诱饵肽的方法来消耗细胞内 FLIL33，从而耗尽MIL33的来源并同时减弱FLIL33的独立影响。 FLIL33 消耗的体内功效的实现将提供对 IL-33 的新认识。病理生物学并开始开发精确、创新的治疗方式。