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.

项目总结 胰腺导管腺癌(PDAC)的特点是对目前的治疗方法耐药，高度 结缔组织增生和免疫抑制的肿瘤微环境(TME)。这项科学提案的重点是 作为KRAS下游转录因子的环磷酸腺苷反应元件结合蛋白1(CREB) 促进疾病侵袭性和低存活率。CREB导联的遗传丢失或药物抑制 在多个小鼠PDAC模型中显著减轻PDAC肿瘤负担并提高存活率。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CREB调节的关键免疫调节细胞因子，可能是肿瘤的旁分泌介质。 巨噬细胞在TME中的串扰。我们假设，靶向CREB-LIF信号轴可以重塑 明显冲击肿瘤相关巨噬细胞免疫抑制TME()S 以T细胞为基础的适应性抗肿瘤免疫反应，提高检查点免疫治疗的疗效。 根据我们的初步数据，CREB调控的LIF信号的关键作用将通过 三个具体目标。1)阐明CREB调控PDAC中LIF的分子机制及其影响 致癌作用。在这里，我们将评估CREB介导的肿瘤细胞来源的LIF的调节和功能作用 通过探索核质定位、与转录复合体的相互作用和基因调控 网络。我们利用CRISPR/Cas9基因组鉴定了肿瘤细胞生长中CREB-LIF轴的需求 用CREB编辑活体胰腺组织并评价LIF相对CREB的表达 患者PDAC组织、患者来源的异种移植物和原发肿瘤衍生有机物。2)确定如何 巨噬细胞中依赖CREB的LIFR信号影响肿瘤免疫格局。我们假设 CREB调节LIF释放并促进向促肿瘤M2样细胞的渗透/极化 巨噬细胞中激活STAT3的表型。我们将确定CREB调节的LIF释放的效果 用CREB开启或关闭对LIF基因敲除肿瘤细胞的作用及评价CREB调控的影响 LIF信号在髓系特异性基因敲除PDAC进展中的获得性免疫反应和TME中的作用 LIFR和STAT3的模型，以及3)CREB抑制和传统免疫治疗方法 减少肿瘤生长，提高总体存活率。我们假设肿瘤细胞固有的CREB-LIF 激活是推动胰腺TME免疫抑制的关键开关，免疫抑制可以协同 免疫检查点抑制。我们将使用CREB抑制剂和检查点抑制剂来确定 治疗和研究遗传小鼠模型、患者来源的异种移植和 有机化合物以进一步补充这些临床前研究。总而言之，这些结果将提供新的治疗方法 减少这种疾病死亡率的策略。此外，这些研究可广泛适用于 CREB-LIF信号发生改变的多种其他恶性肿瘤。