Investigating the Ability of Human Blood Neutrophils to Kill Cancer

研究人类血液中性粒细胞杀死癌症的能力

• 批准号: 10648774
• 负责人: Jonathan S Reichner
• 金额: $ 8.2万
• 依托单位: RHODE ISLAND HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-05 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10648774
• 关键词: 3-Dimensional; Adherent Culture; Adhesions; Adjuvant; Adjuvant Chemotherapy; Agonist; Antibodies; Antibody Therapy; Antitumor Response; Apoptosis; Biomimetics; Breast Cancer Cell; Breast Cancer Patient; Budgets; CD18 Antigens; Cell membrane; Cell surface; Cells; Cellular Spheroids; Cessation of life; Clinic; Clinical; Collaborations; Complement; Cytotoxic T-Lymphocytes; Data; Disease; Effectiveness; Effector Cell; Elasticity; Epidermal Growth Factor Receptor; Exhibits; Extracellular Matrix; Family; Fibroblasts; Fibroins; Gel; Genetic; Greek; Host Defense; Human; ITGAM gene; ITGB2 gene; Immune; Immune response; Immunotherapy; Infection; Integrins; Invaded; Laboratories; Learning; Lectin; MDA MB 231; Macrophage; Macrophage-1 Antigen; Malignant Neoplasms; Mediating; Medical; Modeling; Natural Killer Cells; Necrosis; Neutrophil Infiltration; Organ; Organ failure; Phagocytosis; Physiological; Protocols documentation; Psychological reinforcement; Publishing; Puncture procedure; Regulatory Element; Sepsis; Silk; Site; Structure; Synapses; System; Testing; Therapeutic; Tissues; Tumor Antibodies; Work; anti-cancer; antibody-dependent cell cytotoxicity; cancer cell; cancer immunotherapy; cell killing; clinically relevant; cytotoxic; druggable target; efficacy evaluation; experimental study; insight; interest; live cell imaging; malignant breast neoplasm; migration; neoplastic cell; neutrophil; physical property; receptor; three dimensional cell culture; triple-negative invasive breast carcinoma; tumor; tumor progression

Abstract Neutrophils are not often considered as cancer killing immune cells in the same way that natural killer cells, cytotoxic T cells and macrophages. However, the inherent capability of neutrophils to be cytotoxic is clinically evident in diseases of hyperinflammation (such as sepsis) where activated neutrophils contribute to organ failure by destroying otherwise healthy cells and tissues. Therefore, neutrophils possess all the cytolytic machinery needed to be destructive to host tissues and we propose that includes cancer. What is lacking is a clearly defined targeting mechanism that can focus the cytotoxic potential of neutrophils towards cancer cells in a specific and controlled manner. This proposal will present published and preliminary evidence that neutrophils can indeed be successfully targeted to cancer cells leading to a newly described non-apoptotic cancer killing mechanism called trogoptosis. Trogoptosis (Greek: trogo; “gnaw”) is a newly described effector mechanism that brings neutrophils into the discussion of tumor killing immune cells. Trogoptosis is a form of antibody-dependent cellular cytotoxicity (ADCC) that triggers neutrophils to destroy antibody-opsonized tumor targets by forming a transient receptor-mediated synapse between the neutrophil and the tumor target. Killing occurs as a result of the neutrophil gnawing at the tumor plasma membrane and dismembering it into chunks, and is distinct from apoptosis and necrosis. Neutrophils internalize these pieces of tumor cells by phagocytosis and migrate away. Trogoptosis is dependent on the neutrophil beta2 integrin Complement Receptor 3 (CR3; CD11b/CD18; Mac-1). Antibody blockade of CR3, or neutrophils from families with genetic absence of CR3, obviate trogoptosis. Given the relative recency of this discovery, there remains much to be learned about this effector mechanism. A highly selected experimental plan is offered to fit within the budget and timeframe of an R03 yet allow rigorous testing of the hypothesis that neutrophil trogoptosis is a function that can be enhanced by agonists of neutrophil CR3 and augment the efficacy of anti-cancer antibody therapy. This R03 would develop a nascent collaboration between two laboratories with a track record in the use of biomimetic experimental systems to study breast cancer invasion and the function of human neutrophils. The experimental approach will use fibrous gels of varying stiffnesses and ECM composition that reflect differences that exist among various bodily tissues and organs. MDA-231 TNBC will be modeled as single invasive cells (Specific Aim 1) and as 3D spheroids (Specific Aim 2) in testing the trogoptoptic killing by human neutrophils introduced as tumoricidal effector cells. Additional work will determine if neutrophil CR3 is a druggable target that can enhance the effectiveness of neutrophil-dependent tumor killing and thereby potentially serve as an adjuvant to anti-tumor antibody therapy in the clinic. Our overall objective is to demonstrate that human neutrophils can be stimulated to function effectively and specifically as tumoricidal immune cells.

抽象的 中性粒细胞通常不被认为是癌症，它会像自然杀伤细胞那样杀死免疫细胞， 细胞毒性 T 细胞和巨噬细胞。然而，中性粒细胞固有的细胞毒性能力在临床上是不确定的。 在过度炎症疾病（如脓毒症）中很明显，其中活化的中性粒细胞会导致器官衰竭 通过破坏其他健康的细胞和组织。因此，中性粒细胞拥有所有的细胞溶解机制 需要对宿主组织具有破坏性，我们建议包括癌症。缺少的是一个明确的定义 靶向机制可以将中性粒细胞的细胞毒性潜力集中在特定的和特定的癌细胞上 受控方式。该提案将提供已发表的初步证据，表明中性粒细胞确实可以 成功靶向癌细胞，从而产生新描述的非凋亡癌症杀伤机制 称为斜视。 Trogoptosis（希腊语：trogo；“啃”）是一种新描述的效应机制，它可以带来 中性粒细胞进入肿瘤杀伤免疫细胞的讨论。 Trogooptosis 是抗体依赖性细胞的一种形式 细胞毒性（ADCC），触发中性粒细胞通过形成短暂的抗体来破坏抗体调理的肿瘤靶标 中性粒细胞和肿瘤靶标之间受体介导的突触。杀戮的发生是由于 中性粒细胞啃咬肿瘤质膜并将其肢解成块，这与 细胞凋亡和坏死。中性粒细胞通过吞噬作用将这些肿瘤细胞碎片内在化并迁移走。 Trogoptosis 依赖于中性粒细胞 β2 整合素补体受体 3（CR3；CD11b/CD18；Mac-1）。 CR3 或来自基因缺失 CR3 的家族的中性粒细胞的抗体阻断可避免 trogoptosis。给定 尽管这一发现相对较新，但关于这种效应机制仍有很多需要了解的地方。 提供精心挑选的实验计划，以适应 R03 的预算和时间范围，但允许 严格测试中性粒细胞凋亡是一种可以通过激动剂增强的功能的假设 中性粒细胞 CR3 并增强抗癌抗体治疗的功效。这 R03 将开发一个新生的 两个在使用仿生实验系统方面拥有良好记录的实验室之间的合作 研究乳腺癌侵袭和人类中性粒细胞的功能。实验方法将使用纤维 不同硬度和 ECM 成分的凝胶反映了不同身体组织之间存在的差异 和器官。 MDA-231 TNBC 将被建模为单个侵袭细胞（具体目标 1）和 3D 球体 （具体目标 2）测试作为杀肿瘤效应细胞引入的人类中性粒细胞的视视杀伤作用。 额外的工作将确定中性粒细胞 CR3 是否是可增强药物有效性的药物靶点 中性粒细胞依赖性肿瘤杀伤，因此有可能作为抗肿瘤抗体治疗的佐剂 在诊所里。我们的总体目标是证明人类中性粒细胞可以被刺激发挥功能 有效且特异地作为杀肿瘤免疫细胞。