p27pTpT drives cancer-promoting inflammation and shapes the tumor microenvironment (TME) toward a more tumor-permissive state in vivo

p27pTpT 驱动促癌炎症，并将肿瘤微环境 (TME) 塑造为更适合体内肿瘤生长的状态

• 批准号: 10744370
• 负责人: SeyedehFatemeh Razavipour
• 金额: $ 3.65万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-17 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10744370
• 关键词: ANGPTL4 gene; ATAC-seq; Address; Adipocytes; Affect; Automobile Driving; Biological Assay; Biological Response Modifiers; Blood; Breast Cancer Risk Factor; Breast cancer metastasis; C-terminal; CCL2 gene; CD8-Positive T-Lymphocytes; Cancer Etiology; Cancer Research Project; Cell Cycle; Cell Cycle Inhibition; Cells; Cessation of life; ChIP-seq; Chromatin; Chronic; Combined Modality Therapy; Development; Doctor of Philosophy; Drug resistance; Endocrine; Environment; Estrogen Receptors; Flow Cytometry; Gene Expression; Genes; Genetic Transcription; Goals; Grant; High Fat Diet; Human; IL6 gene; Immune; Immune Evasion; Immune checkpoint inhibitor; Immunologist; Immunology; Immunotherapeutic agent; Infiltration; Inflammation; Inflammation Mediators; Inflammatory; Interleukin-6; Investigation; Link; Macrophage; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Methods; Molecular; Molecular Biology; Mus; Natural Killer Cells; Neoplasm Metastasis; Obese Mice; Obesity; Oncogenes; Oncogenic; PIK3CG gene; Paper; Pathway interactions; Persons; Phase; Phosphorylation; Phosphotransferases; Play; Postdoctoral Fellow; Prevalence; Property; RELA gene; Recurrent Malignant Neoplasm; Regulatory T-Lymphocyte; Research; Research Methodology; Research Personnel; Research Project Grants; Resistance; Role; STAT3 gene; Shapes; Testing; Thinness; Training; Tumor Escape; Tumor Promotion; Tumor Suppressor Proteins; VEGFA gene; Woman; Work; anti-PD-1; cancer cell; cancer recurrence; cancer stem cell; cancer therapy; chemotherapy; cytokine; diet-induced obesity; effective therapy; embryonic stem cell; hormone therapy; immune cell infiltrate; immunoregulation; in vivo; inhibitor; knock-down; malignant breast neoplasm; mimetics; mortality; neoplastic cell; novel; obese person; p65; peripheral blood; permissiveness; pre-doctoral; prevent; programmed cell death ligand 1; programs; promoter; recruit; self-renewal; single-cell RNA sequencing; stem cell expansion; stem cells; targeted treatment; therapy resistant; training opportunity; transcription factor; transcriptome sequencing; tumor; tumor immunology; tumor initiation; tumor microenvironment

Breast Cancer (BC) is the leading cause of cancer-related death in women. Therapy resistance is a major limiting factor. Therefore, the long-term goal of this grant is to elucidate novel molecular mechanisms underlying therapy resistance. BC stem cells (CSCs) may mediate treatment resistance in part by altering the tumor microenvironment (TME) to promote immune evasion. p27 plays a dual role: as a tumor suppressor, it inhibits cell cycle; and as an oncogene, it promotes metastasis when phosphorylated by PI3K activated kinases on its C-terminal at T157 and T198 (p27pTpT). p27pTpT acts as a transcriptional regulator of cJun, driving EMT/pro-metastatic gene profiles. My thesis work showed it also coactivates STAT3 to upregulate CSCs and cancer promoting-inflammation. My overarching hypothesis is that p27pTpT co-activates STAT3 gene profiles to expand cancer stem cells and promote immune evasion. My thesis work in AIM1.1 made the novel observation that p27pTpT upregulates stem cell properties including sphere formation and embryonic stem cell transcription factors (ES-TFs, MYC, OCT4, NANOG, and KLF4), by co-activating STAT3. My ChIP- seq/RNA-seq showed p27 recruits STAT3 to gene promoters to induce broad profiles of p27/STAT3 co-targets, including MYC and JAG1, to increase tumor-initiating stem cells (TISC) in vivo. I showed p27pTpT-driven CSC expansion is in part mediated through the novel p27/STAT3 co-target gene, ANGPTL4. In the F99, AIM1.2, pursues the hypothesis that p27pTpT/STAT3 promotes tumor immune evasion by re-shaping the TME. I will expand my research into cancer immunology which provides me with an outstanding training opportunity. My ATAC-seq showed p27pTpT increases chromatin accessibility to co-recruit p27/STAT3 at promoters of oncogenic, proinflammatory genes (IL-6, NF-κB (RELA), VEGFA) and the mediator of immune evasion, PDL1. p27pTpT also increased NF-κB (p65), IL6 proteins and secreted CCL2. Next, I will test if p27pTpT drives cancer-promoting inflammation and shapes the TME toward a more tumor-permissive state in vivo. First, I will identify how p27pTpT modulates both tumor cell and immune cell signatures and cytokine expression by single cell RNA-seq of BC and Luminex assays of cytokines in peripheral blood of tumor bearing mice in vivo. I will also test how p27pTpT might drive immune evasion by modulating CD4, CD8 T cell and NK cell infiltration in TME and test if p27pTpT recruits MDSCs, Tregs and increases the M2/M1 macrophage ratio in the TME by IHC and by flow cytometry. In AIM2, my K00 addresses mechanisms underlying the greater prevalence and mortality of breast cancer in obese women. My K00 will investigate if p27pTpT promotes the chronic inflammation of obesity by cooperating with the estrogen receptor (ER) to activated NFB/STAT3 to promote immune evasion and endocrine therapy resistance. I will further investigate if dual therapy with ER blockade and checkpoint inhibitors can reverse endocrine resistance in breast cancers in obese hosts. The F99/K00 will help my transition from predoctoral to postdoctoral research and, ultimately, into an independent investigator.

乳腺癌（BC）是女性癌症相关死亡的主要原因。治疗抵抗是一个主要的 限制因素因此，该基金的长期目标是阐明新的分子机制 潜在的治疗抵抗BC干细胞（CSC）可以部分地通过改变细胞的增殖来介导治疗抗性。 肿瘤微环境（TME），以促进免疫逃避。p27具有双重作用：作为肿瘤抑制因子， 抑制细胞周期;并且作为癌基因，当被PI 3 K激活磷酸化时促进转移 在其C-末端T157和T198（p27 pTpT）处存在激酶。p27 pTpT作为cJun的转录调节因子， 驱动EMT/促转移基因谱。我的论文工作表明，它也协同激活STAT 3， CSC和癌症促进炎症。我的总体假设是p27 pTpT共激活STAT 3 基因图谱来扩增癌症干细胞和促进免疫逃避。我的论文工作在AIM1.1中， p27 pTpT上调干细胞特性新观察，包括球体形成和胚胎 干细胞转录因子（ES-TF，MYC，OCT 4，NANOG和KLF 4），通过共激活STAT 3。我的芯片- seq/RNA-seq显示p27将STAT 3募集到基因启动子以诱导p27/STAT 3共靶点的广泛分布， 包括MYC和JAG 1，以增加体内肿瘤起始干细胞（TISC）。我展示了p27 pTpT驱动的CSC 扩增部分通过新的p27/STAT 3共靶基因ANGPTL 4介导。在F99，AIM1.2， p27 pTpT/STAT 3通过重塑TME促进肿瘤免疫逃避。我会 扩大我对癌症免疫学的研究，这为我提供了一个出色的培训机会。我 ATAC-seq显示，p27 pTpT增加了染色质可及性，以在p27/STAT 3的启动子处共募集p27/STAT 3。 致癌、促炎基因（IL-6、NF-κB（RELA）、VEGFA）和免疫逃避介导因子PDL 1。 p27 pTpT还增加NF-κB（p65）、IL 6蛋白和CCL 2的分泌。接下来，我将测试p27 pTpT是否驱动 促进癌症的炎症，并使TME在体内形成更允许肿瘤的状态。首先我会 鉴定p27 pTpT如何通过单一的细胞因子调节肿瘤细胞和免疫细胞特征以及细胞因子表达 BC的cell RNA-seq和体内荷瘤小鼠外周血中细胞因子的Luminex测定。我会 还测试了p27 pTpT如何通过调节CD 4、CD 8 T细胞和NK细胞浸润来驱动免疫逃避。 TME和测试p27 pTpT是否通过以下方式募集MDSC、TcM并增加TME中的M2/M1巨噬细胞比率： 通过IHC和流式细胞术。在AIM 2中，我的K 00解决了更高患病率的潜在机制， 肥胖妇女乳腺癌的死亡率。我的K 00将研究p27 pTpT是否促进慢性 肥胖炎症通过协同雌激素受体（ER）激活NF κ B B/STAT 3促进 免疫逃避和内分泌治疗抗性。我将进一步研究ER阻断的双重治疗是否 检查点抑制剂可以逆转肥胖宿主乳腺癌的内分泌抵抗。F99/K 00将 帮助我从博士前研究过渡到博士后研究，并最终成为一名独立的研究者。