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的顺式和反式调节元件，这些元件在基因转录到翻译的不同阶段调节基因的活性。在全基因组关联研究中，基因组非编码区或调节区的改变与自身免疫性1型糖尿病(T1D)有关，提示基因或基因产物表达的改变可能在功能性免疫失调中发挥重要作用。我们已经在NLRP3的3个Ꞌ-非翻译区(非翻译区)上发现了多个转录后调控元件，NLRP3是代谢功能障碍的细胞质固有免疫传感器，影响NLRP3mRNA的稳定性，并与T1D风险相关。NLRP3的激活导致一个称为炎症体的大型寡聚信号复合体的组装，该复合体促进一种称为下垂的炎性细胞死亡，并产生生物活性的IL-1β，这是一种与T1D的病理生理有关的促炎细胞因子。在NOD(非肥胖糖尿病)小鼠模型中，NLRP3缺乏通过抑制IL-1β的产生和减少致病T细胞向胰岛的迁移来保护T1D的发展，提示NLRP3炎症体在T1D发病中起关键作用。虽然这一组相关事件被很好地描述了，但对控制NLRP3表达并为炎症体激活设定阈值的转录后机制以及与T1D发展相关的病理事件知之甚少。我们的遗传数据证明，人类NLRP3基因3Ꞌ-UTR单核苷酸多态性功能性增加NLRP3mRNA与T1D风险显着相关，提示NLRP3增加可能在疾病发展中发挥作用。我们发现NLRP3基因还有两个多聚腺苷酸化位点，它们要么编码一个稳定的短3Ꞌ-非编码区，要么编码一个不太稳定的长3Ꞌ-非编码区。较长的3Ꞌ-非编码区含有几个顺式作用的调控基序，如富含AU的元件、miRNA结合位点和翻译元件的伽马干扰素抑制因子，这些元件被认为可以抑制蛋白质的表达。根据这些数据，我们假设优先使用短的3‘-UTR而不是长的3’-UTR增加了NLRP3的基因剂量，从而使细胞对NLRP3激活剂敏感，降低了T1D中NLRP3炎症体激活的阈值。在这项资助中，我们将在NOD背景下产生携带NLRP3的短和长3‘-非编码区变体的小鼠，目的是通过交替使用3个Ꞌ-非编码区来解剖NLRP3的基因剂量对T1D在体内的发展和进展的影响。我们的研究将建立基础的小鼠模型来研究转录后调控在T1D发生中的作用，并将对理解遗传变异引起的表达变化如何影响T1D等复杂自身免疫性疾病的发展具有更广泛的意义。

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？

• DOI: 10.1016/j.cels.2022.11.009
• 发表时间: 2022
• 期刊: Cell systems
• 影响因子: 9.3
• 作者: Kuchina,Anna;Yang,Jason;Aldridge,Bree;Janes,KevinA;Subramanian,Naeha;Krogan,NevanJ;Bouhaddou,Mehdi;Einav,Shirit;Papin,Jason;Germain,RonaldN
• 通讯作者: Germain,RonaldN