Impact of CREB-driven mechanism in shaping the tumor-immune landscape

CREB ​​驱动机制对塑造肿瘤免疫景观的影响

• 批准号: 10586923
• 负责人: Nagaraj S. Nagathihalli
• 金额: $ 32.48万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2027-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10586923
• 关键词: Affect; Automobile Driving; Binding; Biological Response Modifiers; CD8-Positive T-Lymphocytes; CRISPR/Cas technology; Cell Line; Cells; Chemotaxis; Clinical; Complement; Complex; Cyclic AMP-Responsive DNA-Binding Protein; Data; Development; Disease; Fibrosis; Genetic; Genetic Models; Genetic Transcription; Genetically Engineered Mouse; High-Throughput Nucleotide Sequencing; Human; Immune; Immune checkpoint inhibitor; Immunophenotyping; Immunosuppression; Immunotherapeutic agent; Immunotherapy; Impairment; Infiltration; Inflammation; Interleukin-6; KRAS2 gene; Knock-out; LIF gene; Laboratories; Macrophage; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Mediator; Modeling; Molecular; Mus; Myelogenous; Myeloid-derived suppressor cells; Nature; Neoplasm Metastasis; Neoplasms; Oncogenic; Organoids; Pancreas; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Patients; Phenotype; Primary Neoplasm; Production; Prognosis; Receptor Signaling; Regulation; Regulatory T-Lymphocyte; Resistance; Role; STAT3 gene; Safety; Sampling; Shapes; Signal Transduction; T-Cell Activation; T-Lymphocyte; Therapeutic; Time; Tissues; Tumor Burden; Tumor Promotion; Tumor-associated macrophages; Up-Regulation; Work; adaptive immune response; anti-tumor immune response; attenuation; cell growth; checkpoint inhibition; checkpoint therapy; chemotherapeutic agent; clinically relevant; cytokine; effective therapy; gene regulatory network; genome editing; humanized mouse; immunomodulatory therapies; immunoregulation; improved; in vivo; inhibitor; insight; leukemia inhibitory factor receptor; mortality; mouse model; neoplastic cell; novel therapeutic intervention; pancreatic ductal adenocarcinoma model; pancreatic neoplasm; paracrine; patient derived xenograft model; pharmacologic; pre-clinical; preclinical study; promoter; recruit; release factor; response; synergism; therapy resistant; transcription factor; transcription regulatory network; treatment strategy; tumor; tumor growth; tumor microenvironment; tumor-immune system interactions; tumorigenesis

PROJECT SUMMARY Pancreatic ductal adenocarcinoma (PDAC) is characterized by resistance to current therapies, a high degree of desmoplasia, and an immunosuppressive tumor microenvironment (TME). This scientific proposal focuses on cyclic AMP Response Element Binding protein 1 (CREB) as a transcriptional factor downstream of KRAS that promotes disease aggressiveness and poor survival. Genetic loss or pharmacological inhibition of CREB leads to significant attenuation of PDAC tumor burden and improved survival in multiple murine PDAC models. Utilizing high throughput sequencing approaches, we have identified tumor cell-derived leukemia inhibitory factor (LIF) as a key CREB-regulated immunomodulatory cytokine, serving as a possible paracrine mediator of tumor- macrophage crosstalk in the TME. We hypothesize that targeting the CREB-LIF signaling axis can remodel the immunosuppressive TME by significantly impacting tumor-associated macrophages (TAM)s and reinvigorating the T cell-based adaptive anti-tumor immune response to improve the efficacy of checkpoint immunotherapy. Based on our preliminary data, the critical role of the CREB regulated LIF signaling will be investigated through three specific aims. 1) Elucidate the molecular mechanism and impact of CREB regulated LIF in PDAC oncogenesis. Here, we will assess the CREB-mediated regulation and functional roles of tumor cell-derived LIF by exploring the nucleo-cytoplasmic localization, interaction with transcriptional complexes, and gene regulatory networks. We identify the CREB-LIF axis requirement in the tumor cell growth using CRISPR/Cas9 genome editing with CREB on or off in vivo pancreas tissues and evaluate the expression of LIF relative to CREB in patient PDAC tissues, patient-derived xenografts, and primary tumor derivative organoids. 2) Determine how CREB-dependent LIFR signaling in macrophages impacts the tumor immune landscape. We hypothesize that the CREB modulates LIF release and promotes TAM infiltration/polarization towards a tumor-promoting M2-like phenotype via activating STAT3 in macrophages. We will determine the effect of CREB-regulated LIF release on TAM activity using CREB on or off with LIF knockout tumor cells and assess the impact of CREB-regulated LIF signaling on the adaptive immune response and TME during PDAC progression in myeloid-specific knockout models of Lifr and Stat3, and 3) Determine if CREB inhibition and conventional immunotherapeutic approaches reduce tumor growth and improve overall survival. We hypothesize that the tumor cell-intrinsic CREB-LIF activation is a pivotal switch that drives immune suppression in the pancreatic TME, which can synergize with immune checkpoint inhibition. We will use CREB inhibitor with checkpoint inhibitor to establish the safety of the therapies and study tumor growth and overall survival in genetic mouse models, patient-derived xenografts, and organoids to further complement these preclinical studies. Together, these results will provide new therapeutic strategies to reduce mortality from this disease. Furthermore, these studies can be broadly applicable to the myriad of other malignancies where CREB-LIF signaling is altered.

项目摘要 胰腺导管腺癌（PDAC）的特征是对当前疗法的抗性，高度 脱木质和免疫抑制肿瘤微环境（TME）。该科学提议着重于 环状AMP响应元件结合蛋白1（CREB）作为KRAS下游的转录因子 促进疾病的侵略性和生存率不佳。遗传丧失或CREB铅的药理抑制 在多种鼠PDAC模型中，PDAC肿瘤负担的显着衰减并改善了生存率。利用 高通量测序方法，我们已经确定了肿瘤细胞衍生的白血病抑制因子（LIF） 作为关键CREB调节的免疫调节细胞因子，是肿瘤的旁分泌介质 TME中的巨噬细胞串扰。我们假设针对Creb-lif信号轴可以重塑 通过显着影响肿瘤相关的巨噬细胞（TAM）和重新活化，免疫抑制TME 基于T细胞的适应性抗肿瘤免疫反应，以提高检查点免疫疗法的功效。 根据我们的初步数据，将通过CREB调节的LIF信号的关键作用 三个具体目标。 1）阐明了PDAC中CREB调节的LIF的分子机制和影响 肿瘤发生。在这里，我们将评估CREB介导的调节和肿瘤细胞衍生LIF的功能作用 通过探索核总质定位，与转录复合物的相互作用和基因调节 网络。我们使用CRISPR/CAS9基因组确定肿瘤细胞生长中的CREB-LIF轴需求 用CREB打开或关闭体内胰腺组织的编辑，并评估LIF相对于CREB的表达 患者PDAC组织，患者衍生的异种移植物和原发性肿瘤衍生物器官。 2）确定如何 巨噬细胞中依赖CREB的LIFR信号会影响肿瘤免疫景观。我们假设这一点 CREB调节LIF释放并促进TAM浸润/极化向促进肿瘤的M2样 通过激活STAT3在巨噬细胞中的表型。我们将确定CREB调节的LIF释放的效果 使用CREB与LIF基因敲除肿瘤细胞打开或关闭TAM活性，并评估CREB调节的影响 髓样特异性敲除PDAC进展过程中自适应免疫反应和TME的LIF信号传导 LIFR和STAT3的模型，以及3）确定CREB抑制和常规免疫治疗方法是否 减少肿瘤生长并改善总体生存率。我们假设肿瘤细胞中的Creb-lif 激活是一个关键开关，可在胰腺TME中驱动免疫抑制作用，可以与 免疫检查点抑制。我们将使用带有检查点抑制剂的CREB抑制剂来确定 疗法和研究肿瘤的生长以及遗传小鼠模型，患者衍生异种移植物和 器官进一步补充这些临床前研究。这些结果将共同提供新的治疗 降低这种疾病死亡率的策略。此外，这些研究可以广泛适用于 其他的恶性肿瘤发生了改变。