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实质性进展： a) 评估 C3 在新发现的细胞内补体系统（ICS 或“复合体”）中的作用， 包括其通过 CD46 信号传导的调节，b) 开发一种测定法来定量生物活性形式 C3 称为 C3(H2O)，c) 识别大多数细胞类型常见的受体介导的 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 反应 介导的攻击。我们当前和持续的目标是发表描述人类受体的结果 C3(H2O) 以及研究表明肺移植的 C3 表达在同种免疫中减少 介导的伤害。我们还在进一步描述复合体激活和调节机制 RNA 和蛋白质水平，并将我们的分析扩展到人外周血吞噬细胞、上皮细胞和， 特别是内皮细胞群。进一步的目标是利用体外实验来剖析 ICS 的作用 具有从肺组织供体分离的原代人类细胞和相应的人类细胞系的模型系统， 以及小鼠感染模型中。因此，我们建议继续开发和使用模型系统 获得关于细胞类型的复合体在正常稳态中如何发挥作用的详细图谱及其 对细胞损伤和恶性肿瘤的反应。此类分析可能会引发关于如何 该系统的功能障碍与人类疾病相关。 该提案的一个重要目标涉及肿瘤学。除了作为抑制剂的作用外 CD46 是宿主细胞上补体激活的关键成分，也是复合体的关键组成部分。 恶性转化和癌症免疫治疗的参与者。一方面，CD46 在 许多肿瘤，但另一方面，它是治疗性溶瘤病毒的目标，使用它作为对接 机制。我们为这些研究做出了贡献，现在计划剖析其过度表达的原理 及其对复合体的影响，从与 Siteman Cancer 合作的多发性骨髓瘤开始 中心。开发的模型随后将扩展到其他类型的癌症。 总之，我们的提案借鉴了我们对这一领域的长期承诺的力量 补充我们在定义机制和参与者方面的研究和雄心勃勃的事业 新发现的ICS。我们对这种资助机制特别感兴趣，因为它的灵活性。这 提案不仅利用了 PI 的专业知识，还利用了一支由优秀人才组成的团队 通过创建更具创新性的模型系统并获得 全面的地图，扩展了我们与 ICS“运作”相关的知识库。