Project 3: Impact of tumor genetics on PDAC immunobiology and responses to macrophage-targeted immunotherapy

项目 3：肿瘤遗传学对 PDAC 免疫生物学的影响以及对巨噬细胞靶向免疫治疗的反应

• 批准号: 10187127
• 负责人: EDGAR G. ENGLEMAN
• 金额: $ 44.33万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10187127
• 关键词: Affect; Agonist; Antibodies; Antigen-Presenting Cells; Automobile Driving; Base Composition; Biological Assay; CDKN2A gene; Cell Line; Cells; Complement; DNA Sequence Alteration; DNA Sequencing Facility; Data; Development; Disease Progression; Duct (organ) structure; Ductal Epithelial Cell; Environment; Enzyme-Linked Immunosorbent Assay; Exhibits; Frequencies; Genetic; Genetically Engineered Mouse; Genomics; Genotype; Goals; Granulocyte-Macrophage Colony-Stimulating Factor; Growth; Human; Immune; Immune response; Immune system; Immunobiology; Immunologics; Immunotherapy; KRAS2 gene; Lesion; Ligands; MADH4 gene; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Methods; Modeling; Molecular; Mus; Mutation; Organism; Pancreatic Ductal Adenocarcinoma; Pancreatic duct; Pattern recognition receptor; Pharmacology; Property; Quantitative Reverse Transcriptase PCR; Resistance; Role; Signal Pathway; Specimen; System; T-Cell Activation; T-Lymphocyte; TP53 gene; Testing; Therapeutic; Transplantation; Treatment Efficacy; Tumor-Derived; Tumor-associated macrophages; Tumor-infiltrating immune cells; Variant; Work; analytical tool; antitumor effect; base; cancer cell; cell type; cytokine; driver mutation; effective therapy; exome sequencing; high dimensionality; immune checkpoint blockade; immune function; immunoregulation; improved; macrophage; molecular subtypes; mouse dectin-2; mouse model; mutational status; neoplastic cell; neutralizing antibody; pancreatic ductal adenocarcinoma model; patient population; programs; receptor; recruit; response; single-cell RNA sequencing; targeted treatment; therapy resistant; transplant model; tumor; tumor growth; tumor microenvironment

ABSTRACT (Project 3) Pancreatic ductal adenocarcinoma (PDAC), an aggressive malignancy that is poorly responsive to treatment, is characterized by a prominent infiltration of immune cells. In particular, macrophages are abundant within the tumor microenvironment (TME) in PDAC and contribute to disease progression and treatment resistance. However, the factors affecting the recruitment and distribution of immune cells, including macrophages, in PDAC remain largely unknown. Recently, our collaborators in Project 1 found that tumors differing in their Trp53 status exhibited different immune profiles. Our collaborators further observed that the cell type of origin (acinar or ductal) influenced the tumor molecular subtype (classical or basal-like), which is also known to be shaped by the immune composition of the tumor. We postulate that somatic alterations in cancer cells, the cell type of origin, and tumor molecular subtype influence the immune landscape within the TME and throughout the host during PDAC progression. To test this hypothesis, we will determine immune cell frequency, distribution, and function in the TME and throughout the host using high-dimensional analytical tools (CODEX and CyTOF) on genetically engineered mouse models (GEMMs) of PDAC that vary in their driver mutations and cell type of origin (developed by Project 1). Additionally, we will use molecular methods to identify immunomodulatory factors regulating immune cell recruitment in the context of different genetic mutations and cell type of origin, and we will use genetic, pharmacological, and neutralizing antibody-based approaches to confirm the functional effects of these factors in driving PDAC progression. Importantly, we will validate findings from these studies in human PDAC specimens and in cell lines derived from human pancreatic ductal cells. Finally, we recently found that the pattern recognition receptor, Dectin-2, is expressed on tumor- associated macrophages (TAMs) in an aggressive, transplantable model of PDAC. Notably, administration of natural Dectin-2 ligands induces sustained PDAC regression in a T-cell dependent manner by reprogramming immunosuppressive TAMs into immunostimulatory antigen-presenting cells (APCs). We hypothesize that PDAC can be effectively treated by reprogramming Dectin-2+ TAMs into immunostimulatory APCs. We will treat autochthonous tumors arising in the GEMMs developed by Project 1 with Dectin-2 ligands to test this hypothesis. Furthermore, to identify mechanisms by which Dectin-2 stimulation is therapeutically efficacious, we will use CODEX and CyTOF to observe changes to the immune landscape occurring within the TME and throughout the host upon treatment. Molecular approaches will be used to identify immunomodulatory factors responsible for mediating therapeutic efficacy. Together, these studies will improve our understanding of how the immune system is altered during PDAC development in the context of variation in genetic mutations, cell type of origin, and molecular subtype, and in response to TAM reprogramming.

摘要（项目 3） 胰腺导管腺癌（PDAC）是一种对治疗反应不佳的侵袭性恶性肿瘤， 其特点是免疫细胞明显浸润。特别是，巨噬细胞在 PDAC 中的肿瘤微环境 (TME) 有助于疾病进展和治疗抵抗。 然而，影响免疫细胞（包括巨噬细胞）招募和分布的因素 PDAC 仍然鲜为人知。最近，我们在项目 1 中的合作者发现，肿瘤在其 Trp53 状态表现出不同的免疫特征。我们的合作者进一步观察到细胞来源类型 （腺泡或导管）影响肿瘤分子亚型（经典或基底样），这也被认为是 由肿瘤的免疫成分决定。我们假设癌细胞中的体细胞改变 起源类型和肿瘤分子亚型影响 TME 内和整个过程中的免疫景观 PDAC进展期间的宿主。为了检验这个假设，我们将确定免疫细胞频率， 使用高维分析工具（CODEX 和 CyTOF）对 PDAC 基因工程小鼠模型（GEMM）的驱动突变有所不同 和细胞来源类型（由项目 1 开发）。此外，我们将使用分子方法来识别 在不同基因突变的情况下调节免疫细胞募集的免疫调节因子和 来源的细胞类型，我们将使用遗传、药理学和基于中和抗体的方法来 确认这些因素在驱动 PDAC 进展中的功能作用。重要的是，我们将验证 这些研究结果来自于人类 PDAC 标本和源自人类胰腺导管的细胞系 细胞。最后，我们最近发现模式识别受体 Dectin-2 在肿瘤上表达 侵袭性、可移植 PDAC 模型中的相关巨噬细胞 (TAM)。值得注意的是，管理 天然 Dectin-2 配体通过重编程以 T 细胞依赖性方式诱导持续的 PDAC 消退 免疫抑制性 TAM 转化为免疫刺激性抗原呈递细胞 (APC)。我们假设 通过将 Dectin-2+ TAM 重编程为免疫刺激性 APC 可以有效治疗 PDAC。我们将 使用 Dectin-2 配体治疗项目 1 开发的 GEMM 中出现的本土肿瘤，以测试这一点 假设。此外，为了确定 Dectin-2 刺激在治疗上有效的机制， 我们将使用 CODEX 和 CyTOF 观察 TME 内发生的免疫景观变化， 治疗后遍及宿主。分子方法将用于识别免疫调节因子 负责调节治疗效果。总之，这些研究将提高我们对如何 在 PDAC 发育过程中，免疫系统在基因突变、细胞变异的背景下发生改变 起源类型和分子亚型，以及对 TAM 重编程的响应。