Does post-transcriptional control of NLRP3 inflammasome activity impact development of type 1 diabetes?

NLRP3 炎性体活性的转录后控制是否会影响 1 型糖尿病的发展？

• 批准号: 10404093
• 负责人: Naeha Subramanian
• 金额: $ 24.7万
• 依托单位: INSTITUTE FOR SYSTEMS BIOLOGY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-12 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10404093
• 关键词: 3' Untranslated Regions; Affect; Autoimmune; Autoimmune Diseases; Binding; Binding Proteins; Binding Sites; CASP1 gene; Cell Death; Cells; Clinical Trials; Complex; Data; Development; Disease; Elements; Event; Foundations; Functional disorder; Gel; Gene Dosage; Gene Expression; Genes; Genetic; Genetic Transcription; Genetic Variation; Genome; Goals; Grant; Human; Human Genome; Immune; Impairment; Inbred NOD Mice; Inflammasome; Inflammatory; Insulin; Insulin-Dependent Diabetes Mellitus; Interferon Type II; Interleukin-1 Receptors; Interleukin-1 beta; Islet Cell; Islets of Langerhans; Ligands; Link; Mediator of activation protein; Messenger RNA; Metabolic dysfunction; MicroRNAs; Modeling; Molecular; Mus; Non obese; Nucleic Acid Regulatory Sequences; Pancreas; Pathogenesis; Pathogenicity; Pathologic; Pathology; Play; Polyadenylation; Polyadenylation Pathway; Post-Transcriptional Regulation; Predisposition; Production; Proteins; Regulator Genes; Regulatory Element; Research; Role; Signal Transduction; Single Nucleotide Polymorphism; Site; T-Lymphocyte; Th1 Cells; Translations; Untranslated RNA; Untranslated Regions; Variant; base; cytokine; diabetes pathogenesis; diabetes risk; diabetic; gene product; genome wide association study; in vivo; inhibitor; insulin sensitivity; mRNA Stability; microbial; migration; mouse model; protein expression; receptor; sensor

The human genome contains a large number of cis- and trans-acting regulatory elements that modulate gene activity at different points in the progression from gene transcription to translation. Alterations in non-coding or regulatory regions of the genome are linked to autoimmune type 1 diabetes (T1D) in Genome-wide association studies (GWAS), suggesting that changes in gene or gene product expression might play an important role in functional immune dysregulation. We have identified multiple post-transcriptional regulatory elements in the 3Ꞌ-UTR (untranslated region) of NLRP3, a cytosolic innate immune sensor of metabolic dysfunction, that impact NLRP3 mRNA stability and are associated with T1D risk. NLRP3 activation results in assembly of a large oligomeric signaling complex called the inflammasome that promotes a form of inflammatory cell death called pyroptosis and production of bioactive IL-1β, a pro-inflammatory cytokine implicated in the pathophysiology of T1D. In the NOD (non-obese diabetic) mouse model, NLRP3 deficiency protects from development of T1D through impaired production of IL-1β and reduced migration of pathogenic T cells to the pancreatic islets, suggesting a critical role for the NLRP3 inflammasome in T1D pathogenesis. While this set of linked events is well described, little is known about the post-transcriptional mechanisms that control the expression NLRP3 and set the threshold for inflammasome activation and the pathological events associated with development of T1D. Our genetic data document that a single nucleotide polymorphism in the 3Ꞌ-UTR of the human NLRP3 gene that functionally increases NLRP3 mRNA is significantly associated with T1D risk suggesting that an increase in NLRP3 may play a role in disease development. We have found that the NLRP3 gene also has two polyadenylation sites, which either encode for a stable short 3Ꞌ-UTR or a less stable longer 3Ꞌ-UTR. The longer 3Ꞌ-UTR harbors several cis acting regulatory motifs, such as AU-rich elements, miRNA binding sites and gamma interferon inhibitor of translation elements, which are known to suppress protein expression. Based on these data we hypothesize that preferential usage of the short 3'-UTR as opposed to the long 3'-UTR increases the gene dosage of NLRP3 thereby sensitizing cells to NLRP3 activators and lowering the threshold for NLRP3 inflammasome activation in T1D. In this grant we will generate mice carrying the short vs. long 3'-UTR variants of NLRP3 on the NOD background with the goal of dissecting the impact of gene dosage of NLRP3 via alternate 3Ꞌ-UTR usage on the development and progression of T1D in vivo. Our study will generate foundational mouse models to study the role of post-transcriptional regulation in development of T1D and will have broader implications for understanding how expression changes caused by genetic variation impact the development of complex autoimmune diseases like T1D.

人类基因组包含大量顺式和反式作用调节元件，它们在从基因转录到翻译的过程中的不同点调节基因活性，基因组非编码或调节区域的改变与自身免疫性 1 型糖尿病有关。 (T1D) 在全基因组关联研究 (GWAS) 中，表明基因或基因产物表达的变化可能在功能性免疫失调中发挥重要作用，我们在 3Ꞌ​​-UTR 中发现了多个转录后调控元件。 NLRP3 是一种代谢功能障碍的细胞质先天免疫传感器，它会影响 NLRP3 mRNA 的稳定性，并与 T1D 风险相关。NLRP3 激活会导致称为炎症小体的大型寡聚信号复合物的组装，从而促进某种形式的炎症细胞死亡。称为焦亡和生物活性 IL-1β 的产生，IL-1β 是一种与 T1D 病理生理学有关的促炎细胞因子。在糖尿病小鼠模型中，NLRP3 缺陷通过 IL-1β 的产生受损和致病性 T 细胞向胰岛的迁移减少来防止 T1D 的发展，这表明 NLRP3 炎性体在 T1D 发病机制中发挥着关键作用。尽管描述得很清楚，但人们对控制 NLRP3 表达并设定炎症小体激活阈值以及与 T1D 发展相关的病理事件的转录后机制知之甚少。遗传数据表明，人类 NLRP3 基因 3Ꞌ-UTR 中的单核苷酸多态性在功能上增加 NLRP3 mRNA，与 T1D 风险显着相关，这表明 NLRP3 的增加可能在疾病发展中发挥作用。还具有两个聚腺苷酸化位点，它们编码稳定的短 3Ꞌ-UTR 或不太稳定的较长 3Ꞌ-UTR。 3Ꞌ-UTR 包含多个顺式作用调节基序，例如富含 AU 的元件、miRNA 结合位点和翻译元件的 γ 干扰素抑制剂，这些基序已知可抑制蛋白质表达。根据这些数据，我们捕获了短 3' 的优先使用。 -UTR 与长 3'-UTR 不同，增加了 NLRP3 的基因剂量，从而使细胞对 NLRP3 激活剂敏感并降低 NLRP3 炎性体激活的阈值在 T1D 中，我们将在 NOD 背景上生成携带 NLRP3 的短与长 3'-UTR 变体的小鼠，目的是通过交替使用 3Ꞌ-UTR 来剖析 NLRP3 基因剂量对发育和进展的影响。我们的研究将产生基础小鼠模型来研究转录后调控在 T1D 发展中的作用，并将对理解遗传变异引起的表达变化如何影响 T1D 的发展产生更广泛的影响。复杂的自身免疫性疾病，如 T1D。

项目成果

How can systems approaches help us understand and treat infectious disease?

系统方法如何帮助我们理解和治疗传染病？

• 发表时间: 2022-12-21
• 影响因子: 9.3
• 作者: Kuchina, Anna;Yang, Jason;Aldridge, Bree;Janes, Kevin A;Subramanian, Naeha;Krogan, Nevan J;Bouhaddou, Mehdi;Einav, Shirit;Papin, Jason;Germain, Ronald N
• 通讯作者: Germain, Ronald N