Defining the Complosome in Human Cells, Tissues and Disease States

定义人类细胞、组织和疾病状态中的复合体

• 批准号: 10375425
• 负责人: John Atkinson
• 金额: $ 39.38万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-03 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10375425
• 关键词: Address; Autoimmunity; Biological Assay; Biological Models; CD46 Antigen; Cancer Center; Cells; Collaborations; Complement; Complement Activation; Complement Inactivators; Disease; Docking; Endothelial Cells; Epithelial; Functional disorder; Goals; Grant; Homeostasis; Human; Human Cell Line; Immune; Individual; Infection; Inflammatory Response; Injury; Knowledge; Lead; Lung Transplantation; Malignant - descriptor; Malignant Neoplasms; Maps; Mediating; Modeling; Multiple Myeloma; Natural Immunity; Oncology; Oncolytic viruses; Pathway interactions; Phagocytes; Population; Positioning Attribute; Process; Proteins; Publishing; RNA; Research; Role; Signal Transduction; Structure of parenchyma of lung; System; Talents; Tissue Donors; Tissues; body system; cancer immunotherapy; cancer type; cell injury; cell type; complement system; flexibility; human disease; in vitro Model; innovation; insight; knowledge base; mouse model; new therapeutic target; overexpression; peripheral blood; receptor; response; targeted treatment; tumor; uptake

Abstract During the past three and one-half years of R01 grant support, we have made substantial progress in: a) assessing the role of C3 in the newly discovered intracellular complement system (ICS or “complosome”), including its modulation by CD46 signaling, b) developing an assay to quantitate a biologically active form of C3 known as C3(H2O), c) identifying a receptor-mediated C3(H2O) uptake process common to most cell types, and d) determining the ICS response in cells and an organ system (the airway) to ischemic and immune- mediated attack. Our immediate and ongoing goals are to publish our results describing the human receptor for C3(H2O) as well as studies demonstrating that C3 expression by lung transplants is reduced in alloimmune- mediated injury. We are also further delineating complosome activation and regulatory mechanisms at the RNA and protein level and are extending our analyses to human peripheral blood phagocytic, epithelial and, especially, endothelial cell populations. A further goal is to dissect the role of the ICS, utilizing both an in vitro model system with primary human cells isolated from lung tissue donors and corresponding human cell lines, as well as in mouse models of infection. Thus, we propose to continue to develop and employ model systems to obtain a detailed map of how a cell type's complosome functions in normal homeostasis as well as its response to cellular damage and malignancy. Such analyses will likely trigger new insights into how dysfunctions of this system correlate with human diseases. An important goal of this proposal relates to oncology. In addition to its role as an inhibitor of complement activation on host cells and a critical component of the complosome, CD46 is emerging as a key player in both malignant transformation and cancer immunotherapy. On one hand, CD46 is overexpressed on many tumors, yet on the other hand, it is targeted by therapeutic oncolytic viruses that use it as a docking mechanism. We have contributed to these studies and now plan to dissect the rationale of its overexpression and its effect on the complosome, beginning with multiple myeloma in collaboration with the Siteman Cancer Center. The models developed will be extended subsequently to other types of cancers. In summary, our proposal draws on the strength of our long-term commitment to the field of complement research and our ambitious undertakings related to defining the mechanisms and players of the newly discovered ICS. We are especially attracted to this grant mechanism because of its flexibility. This proposal draws not only on the expertise of the PI, but also on a team of talented individuals who are well positioned to continue being leaders the field by creating more innovative model systems and obtaining a comprehensive map that expands our knowledge-base relative to the “workings” of the ICS.

抽象的 在过去三年半的R01赠款支持中，我们在以下方面取得了重大进展： a）评估C3在新发现的细胞内完成系统（ICS或“ APPOSOME”）中的作用， 包括通过CD46信号传导调制其调节 C3称为C3（H2O），C）识别大多数细胞类型共有的接收器介导的C3（H2O）摄取过程， d）确定细胞和器官系统（气道）对缺血和免疫的ICS响应 中介攻击。我们的直接和持续的目标是发布我们的结果，描述了人类接收者的 C3（H2O）以及研究表明，在同种免疫性中降低了肺移植的C3表达 介导的伤害。我们还进一步描述了在 RNA和蛋白质水平，正在将我们的分析扩展到人类外周血吞噬，上皮和， 特别是内皮细胞群体。另一个目标是剖析IC的作用，同时使用体外 与从肺组织供体分离的原代人细胞和相应人类细胞系的模型系统， 以及在小鼠感染模型中。这，我们建议继续开发和员工模型系统 为了获得细胞类型如何在正常体内稳态及其中的详细地图 对细胞损伤和恶性肿瘤的反应。这样的分析可能会引发新的见解 该系统的功能障碍与人类疾病有关。 该提案的一个重要目标与肿瘤学有关。除了作为抑制剂的作用 在宿主细胞上的补体激活和完全集体组的关键成分，CD46正在作为钥匙出现 恶性转化和癌症免疫疗法的玩家。一方面，CD46过表达 许多瘤 机制。我们已经为这些研究做出了贡献，现在计划剖析其过表达的理由 它对与现场癌症合作的多发性骨髓瘤开始，它对联想组的影响 中心。开发的模型将随后扩展到其他类型的癌症。 总而言之，我们的提议取决于我们对该领域的长期承诺的实力 补充研究和我们雄心勃勃的事业与定义的机制和参与者有关 新发现的IC。由于其灵活性，我们特别受到这种赠款机制的吸引。这 提案不仅借鉴了PI的专业知识，还借鉴了一个很好的人才团队 通过创建更具创新的模型系统并获得 相对于IC的“工作”扩展我们的知识基础的综合图。