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）的特征在于对当前疗法的抗性，高度的肿瘤坏死，以及高水平的肿瘤转移。 结缔组织增生和免疫抑制性肿瘤微环境（TME）。这项科学建议的重点是 环AMP反应元件结合蛋白1（CREB）作为KRAS下游的转录因子， 促进了疾病的侵袭性和生存率的下降。CREB导联的遗传丢失或药物抑制 在多种鼠PDAC模型中显著减轻PDAC肿瘤负荷并提高存活率。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） 作为一个关键的CREB调节的免疫调节细胞因子，作为一个可能的旁分泌介导的肿瘤， TME中的巨噬细胞串扰。我们假设，靶向CREB-LIF信号传导轴可以重塑 免疫抑制性TME通过显著影响肿瘤相关巨噬细胞（TAM）和重新激活 基于T细胞的适应性抗肿瘤免疫应答，以提高检查点免疫疗法的疗效。 基于我们的初步数据，CREB调节的LIF信号转导的关键作用将通过以下方式进行研究： 三个具体目标。1)阐明CREB调控LIF在PDAC中的分子机制及其影响 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的LIF的功能作用， 通过探索核质定位，与转录复合物的相互作用，以及基因调控， 网络.我们使用CRISPR/Cas9基因组鉴定了肿瘤细胞生长中CREB-LIF轴的需求 用CREB在体内胰腺组织上或在体内胰腺组织上进行编辑，并评估LIF相对于CREB在胰腺组织中的表达。 患者PDAC组织、患者来源的异种移植物和原发性肿瘤衍生类器官。2)确定如何 巨噬细胞中的CREB依赖性LIFR信号传导影响肿瘤免疫景观。我们假设 CREB调节LIF释放并促进TAM向促肿瘤M2样细胞浸润/极化， 表型通过激活巨噬细胞中的STAT 3来表达。我们将确定CREB调节的LIF释放的影响， 用LIF敲除肿瘤细胞使用CREB开启或关闭对TAM活性的影响，并评估CREB调节的TAM活性的影响。 骨髓特异性基因敲除PDAC进展过程中LIF信号对适应性免疫应答和TME的影响 Lifr和Stat 3的模型，以及3）确定CREB抑制和常规免疫抑制方法是否 减少肿瘤生长并提高总体存活率。我们假设肿瘤细胞内源性CREB-LIF 激活是驱动胰腺TME中免疫抑制的关键开关，其可以与 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 在遗传小鼠模型、患者来源的异种移植物中研究肿瘤生长和总生存期， 类器官以进一步补充这些临床前研究。总之，这些结果将提供新的治疗方法， 降低这种疾病死亡率的战略。此外，这些研究可广泛适用于 CREB-LIF信号传导被改变的无数其他恶性肿瘤。