Anti-Complement Immunotherapy for Pancreatic Cancer

胰腺癌的抗补体免疫治疗

• 批准号: 10751872
• 负责人: Brett I Bell
• 金额: $ 5.27万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-01-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10751872
• 关键词: Alternative Therapies; Anaphylatoxins; Animals; Area; Biological Assay; Bone Marrow; C3AR1 gene; Cancer Etiology; Carbon ion; Cell Membrane Alteration; Cell model; Cells; Cessation of life; Coculture Techniques; Complement; Complement 3a; Complement 3b; Complement 5a; Complement Activation; Complement Membrane Attack Complex; Complement Receptor; Curative Surgery; Data; Dendritic Cells; Dendritic cell activation; Deposition; Development; Disease; Effector Cell; Exclusion; Fibroblasts; Fibrosis; Flow Cytometry; Generations; Goals; Growth; Histologic; Histology; Histopathologic Grade; Human; Immune; Immune Tolerance; Immunofluorescence Immunologic; Immunologics; Immunomodulators; Immunosuppression; Immunotherapeutic agent; Immunotherapy; Impairment; Implant; In Vitro; Infiltration; Irradiated tumor; KRASG12D; Knock-out; Knockout Mice; Linear Energy Transfer; LoxP-flanked allele; Macrophage; Malignant Neoplasms; Malignant neoplasm of pancreas; Mannose Binding Lectin; Measures; Mediating; Modality; Modeling; Mus; Mutation; Neoplasm Metastasis; PET/CT scan; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Patients; Phagocytes; Photons; Population; Production; Proliferating; Protons; Radiation; Radiation therapy; Regulatory T-Lymphocyte; Reporting; Research; Roentgen Rays; Role; Serum; Signal Transduction; Sirius Red F3B; Source; Stains; Survival Rate; T-Lymphocyte; Tamoxifen; Testing; Therapeutic; Treatment Efficacy; Tumor Immunity; Tumor Promotion; Unresectable; Western Blotting; X-Ray Computed Tomography; cancer therapy; complement deficiency; complement pathway; complement system; fluorodeoxyglucose; immune cell infiltrate; immunomodulatory therapies; improved; in vivo imaging system; inhibitor; insight; irradiation; knock-down; migration; mouse model; neoplastic cell; neutralizing antibody; novel; novel therapeutic intervention; pancreatic cancer model; pancreatic ductal adenocarcinoma model; pancreatic tumorigenesis; proton therapy; receptor; receptor expression; response; restraint; single-cell RNA sequencing; small hairpin RNA; small molecule inhibitor; therapy development; translational study; translational therapeutics; tumor; tumor growth; tumor microenvironment; tumorigenesis

PROJECT SUMMARY / ABSTRACT Pancreatic Ductal Adenocarcinoma (PDAC) is an almost uniformly lethal disease, with an overall survival under 10%. Despite therapeutic advances in all arenas of cancer treatment, including immunotherapy, overall survival has not significantly improved in PDAC, representing a critical need for the development of novel therapeutic strategies for this disease. The tumor microenvironment (TME) in PDAC is characterized by an immunosuppressive infiltrate causing T-cell exclusion, and dense stromal desmoplasia. The complement cascade is activated in the PDAC TME and may promote this uniquely challenging TME by signaling to immune cells and fibroblasts expressing complement receptors. Complement activation has previously been demonstrated to promote infiltration of immune cells which contribute to T-cell exclusion in multiple tumor types in a context-dependent fashion. The long-term objectives of this project are to understand microenvironmental mechanisms by which complement promotes oncogenesis in PDAC, and to elucidate the interactions between radiotherapy (RT) and complement blockade in PDAC. A novel autochthonous mouse model of PDAC was generated and crossed with complement deficient C3 knockout (KO) mice to allow for studies which define the role of complement in PDAC. Tumor growth in complement-proficient wild type (WT) PDAC mice and C3 KO PDAC mice will be tracked using serial, PET/CT imaging. Tumors will be assessed histologically from 3-6 months after induction with tamoxifen to determine histologic grade, normal acinar area, and fibrotic area. Flow cytometry will be used to determine the impact of complement on infiltrating immune cells while single-cell RNA-sequencing will provide insight into the development and activation of cancer-associated fibroblasts. We will also assess the role of tumor cell vs. stromally derived C3 with shRNA depletion of C3 in YFP-expressing KPC cells (KPCY) in flank tumor growth studies in WT and C3 KO mice as a second model to confirm these results. To assess the impact of photon, proton, and carbon ion RT on complement activation and immune tolerance we will irradiate KPCY using each radiation modality. Complement deposition assays will be performed by culturing irradiated and non-treated tumor cells in the presence of serum as a complement source, before measuring activation by flow cytometry. Next, dendritic cells will be co-cultured with irradiated and non-irradiated KPCY cells with serum to determine if complement deposition on irradiated tumor cells reduces dendritic cell activation markers measured by flow cytometry. Translational studies will be performed using neutralizing antibodies and small molecule inhibitors to blockade complement signaling alone and with RT in an orthotopic KPCY model. Mice will be treated with neutralizing antibodies and inhibitors to blockade complement signaling and irradiated with an X- strahl Small Animal Radiation Research Platform or a Varian ProBeam proton therapy machine and growth will be tracked by IVIS. Complement activation will be assessed by western blot and immunofluorescence, and flow cytometry will assess differences in infiltrating immune cells post-irradiation.

项目摘要 /摘要 胰腺导管腺癌（PDAC）几乎是致命的疾病，在总体下生存 10％。尽管在所有癌症治疗领域都有治疗性进步，包括免疫疗法，但总体生存 PDAC尚未显着改善，代表了对新型治疗的迫切需求 这种疾病的策略。 PDAC中的肿瘤微环境（TME）的特征是 免疫抑制浸润，导致T细胞排除和密集的基质脱落。补充 Cascade在PDAC TME中被激活，并可能通过发出免疫来促进这种独特的TME 表达补体受体的细胞和成纤维细胞。补体激活以前已经 被证明可以促进免疫细胞的浸润，这些免疫细胞有助于多种肿瘤类型的T细胞排除 以上下文依赖的方式。该项目的长期目标是了解微环境 补体促进PDAC的肿瘤发生的机制，并阐明 PDAC中的放疗（RT）和补体阻滞。 PDAC的一种新型的自围小鼠模型是 与补体不足的C3敲除（KO）小鼠产生并交叉，以允许进行定义的研究 补体在PDAC中的作用。补体野生型（WT）PDAC小鼠和C3 KO的肿瘤生长 PDAC小鼠将使用串行，PET/CT成像跟踪。肿瘤将从组织学评估3-6个月 他莫昔芬诱导后确定组织学等级，正常腺泡区域和纤维化区域。流式细胞仪 将用于确定补体对浸润的免疫细胞的影响，同时单细胞RNA顺序 将洞悉与癌症相关的成纤维细胞的发育和激活。我们还将评估 在表达YFP的KPC细胞（KPCY）中，肿瘤细胞与基质衍生的C3的作用 WT和C3 KO小鼠的侧面肿瘤生长研究是第二种模型，以确认这些结果。评估 光子，质子和碳离子RT对补体激活和免疫耐受的影响我们将照射 使用每种辐射方式KPCY。补体沉积测定将通过培养辐照来进行 在血清存在作为补体来源的情况下，未经处理的肿瘤细胞，然后测量通过 流式细胞仪。接下来，树突状细胞将与辐照和非辐照的KPCY细胞共培养 确定对辐照肿瘤细胞的补体沉积是否会降低树突状细胞活化标记 通过流式细胞仪测量。翻译研究将使用中和抗体和小 分子抑制剂单独阻断补体信号传导，并在原位KPCY模型中使用RT。老鼠会 用中和抗体和抑制剂进行阻断补体信号传导的抑制剂，并用X-进行照射 Strahl小型动物辐射研究平台或Varian Probeam Proton治疗机和生长将 由Ivis跟踪。补体激活将通过蛋白质印迹和免疫荧光评估，流动 细胞仪将评估辐照后浸润免疫细胞的差异。