Evaluating the role of protein self-assembly in the innate immune system

评估蛋白质自组装在先天免疫系统中的作用

• 批准号: 10045772
• 负责人: Alejandro Rodriguez Gama
• 金额: $ 3.1万
• 依托单位: STOWERS INSTITUTE FOR MEDICAL RESEARCH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10045772
• 关键词: Acute; Address; Age; Aging; Alzheimer' s Disease; Atherosclerosis; Biochemical; Biological Assay; Biophysics; Blood Cells; Cell Culture Techniques; Cell Death; Cell Differentiation process; Cell Line; Cells; Chronic; Cytology; Data; Death Domain; Disease; Elderly; Event; Family; Family member; Genes; Goals; Hematopoiesis; Hematopoietic stem cells; Homeostasis; Human; Immune; Immune signaling; Immunologic Memory; Impairment; In Vitro; Infection; Inflammaging; Inflammasome; Inflammation; Inflammatory; Inflammatory Response; Innate Immune System; Lead; Ligands; Measures; Molecular; Myeloid Cells; Myelopoiesis; Nature; Non-Insulin-Dependent Diabetes Mellitus; Organism; Parkinson Disease; Pathogenesis; Pathologic; Pattern recognition receptor; Phase; Phase Transition; Phenotype; Physiological; Polymers; Prions; Process; Production; Property; Proteins; Receptor Activation; Reporter; Research; Research Personnel; Research Project Grants; Risk; Role; Signal Pathway; Signal Transduction; Stimulus; Techniques; Tertiary Protein Structure; Work; age related; aged; autoinflammation; autoinflammatory; base; combat; cytokine; digital; exhaustion; experimental study; fitness; hematopoietic stem cell self-renewal; in vivo; in vivo Model; innate immune function; innovation; insight; member; monocyte; mortality; mortality risk; novel; novel strategies; particle; pathogen; pathogen exposure; pathogenic fungus; polymerization; post-doctoral training; prevent; prion-like; protein complex; protein function; receptor; response; self assembly; self-renewal; senescence; tool

As humans age, levels of proinflammatory cytokines increase, leading to a chronic inflammatory state, known as inflammaging. Inflammation is a hallmark of age-related diseases and increases mortality among the elderly. The molecular mechanisms that underlie inflammaging have not yet been elucidated. Therefore, there is an urgent need to elucidate the molecular mechanisms causing persistent proinflammatory signals, as doing so will accelerate efforts to prevent and treat age-related and autoinflammatory diseases. The overall goal of this proposal is to determine if a specific molecular mechanism contributes to the proinflammatory phenotype of aged monocytes. The data gathered so far reveals that multiple Death Domain proteins that function as key nodes downstream of pathogen recognition receptors are activated through a nucleation-limited assembly mechanism. The self-sustaining nature of this form of assembly should in principle allow them to persist indefinitely and continuously induce monocytes for proinflammatory signaling. The proposed aims will address two important questions: 1) Do protein self-assemblies suffice to establish and maintain a proinflammatory state in monocytes? 2) Are protein self-assemblies involved in the declining production and function of innate immune cells at the hematopoiesis level? In the F99-phase of this proposed research, the researcher aims to identify the molecular substrate that drives a persistent proinflammatory state in monocytes. The implementation of novel tools to measure functionality of protein self-assemblies in vivo, will address whether the assembled states of certain Death Domain proteins are functionally establishing a proinflammatory state in monocytes. In the K00-phase of this proposed research, the researcher will identify a potential mechanism of hematopoietic stem cell (HSC) decay and impaired self-renewal caused by signalosome formation. Specifically, the proposed experiments will determine whether Death Domain protein self-assemblies functionally reprogram HSC to skew the production of myeloid cells preloaded with a proinflammatory signature. To achieve these goals, the researcher will develop and implement novel approaches using cytological, biochemical and gene editing techniques. Completion of these aims will identify molecular substrates of inflammaging and reveal an unexplored role of ordered protein self-assemblies in signaling and cell differentiation. The proposed work is innovative and brings emerging biophysical concepts to the aging and immune senescence fields.

随着人类年龄的增长，促炎细胞因子的水平增加，导致慢性炎症状态，称为炎症。炎症是与年龄相关的疾病的一个标志，会增加老年人的死亡率。炎症的分子机制尚未阐明。因此，迫切需要阐明导致持续促炎信号的分子机制，因为这样做将加速预防和治疗与年龄相关的疾病和自身炎症疾病。该提案的总体目标是确定特定的分子机制是否有助于衰老单核细胞的促炎表型。迄今为止收集的数据表明，作为病原体识别受体下游关键节点的多个死亡域蛋白通过成核限制组装机制被激活。这种组装形式的自我维持性质原则上应该允许它们无限期地持续并持续诱导单核细胞产生促炎信号。拟议的目标将解决两个重要问题：1）蛋白质自组装是否足以在单核细胞中建立和维持促炎状态？ 2）蛋白质自组装是否与造血水平上先天免疫细胞的产生和功能下降有关？在这项拟议研究的 F99 阶段，研究人员旨在确定驱动单核细胞持续促炎状态的分子底物。测量体内蛋白质自组装功能的新工具的实施将解决某些死亡结构域蛋白质的组装状态是否在单核细胞中功能性地建立促炎状态。在这项拟议研究的 K00 阶段，研究人员将确定信号体形成引起的造血干细胞 (HSC) 衰退和自我更新受损的潜在机制。具体来说，拟议的实验将确定死亡结构域蛋白自组装是否对 HSC 进行功能性重编程，以扭曲预载促炎信号的骨髓细胞的产生。为了实现这些目标，研究人员将利用细胞学、生化和基因编辑技术开发和实施新方法。完成这些目标将识别炎症的分子底物，并揭示有序蛋白质自组装在信号传导和细胞分化中尚未探索的作用。拟议的工作具有创新性，为衰老和免疫衰老领域带来了新兴的生物物理学概念。