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。治疗耐药是一个主要原因。 因此，这项资助的长期目标是阐明新的分子机制。 BC 干细胞 (CSC) 可能部分通过改变治疗抵抗来介导治疗抵抗。 肿瘤微环境（TME）促进p27发挥双重作用：作为肿瘤抑制因子。 抑制细胞周期；并且作为癌基因，当被 P​​I3K 激活磷酸化时会促进转移 其 C 末端 T157 和 T198 处的激酶 (p27pTpT) 充当 cJun 的转录调节因子， 我的论文工作表明它还共同激活 STAT3 来上调。 CSC 和癌症促进炎症。我的总体假设是 p27pTpT 共同激活 STAT3。 我的论文工作是在 AIM1.1 中进行的。 p27pTpT 上调干细胞特性（包括球体形成和胚胎）的新观察 干细胞转录因子（ES-TF、MYC、OCT4、NANOG 和 KLF4），通过共同激活 My ChIP-。 seq/RNA-seq 显示 p27 将 STAT3 募集至基因启动子以诱导广泛的 p27/STAT3 共同靶标， 包括 MYC 和 JAG1，以增加体内肿瘤起始干细胞 (TISC)，我发现 p27pTpT 驱动的 CSC。 在 F99、AIM1.2 中，扩增部分是通过新的 p27/STAT3 共同靶基因 ANGPTL4 介导的。 我将提出 p27pTpT/STAT3 通过重塑 TME 促进肿瘤免疫逃避的假设。 将我的研究扩展到癌症免疫学，这为我提供了绝佳的培训机会。 ATAC-seq 显示 p27pTpT 增加了在启动子处共同招募 p27/STAT3 的染色质可及性 致癌、促炎基因（IL-6、NF-κB (RELA)、VEGFA）和免疫逃避介质 PDL1。 p27pTpT 还增加了 NF-κB (p65)、IL6 蛋白和分泌的 CCL2 接下来，我将测试 p27pTpT 是否驱动。 促进癌症的炎症并使 TME 在体内朝着更容易发生肿瘤的状态发展。 确定 p27pTpT 如何通过单一调节肿瘤细胞和免疫细胞特征以及细胞因子表达 我将在体内对荷瘤小鼠外周血中的细胞因子进行 BC 细胞 RNA 测序和 Luminex 测定。 还测试了 p27pTpT 如何通过调节 CD4、CD8 T 细胞和 NK 细胞浸润来驱动免疫逃避 TME 并测试 p27pTpT 是否招募 MDSC、Treg 并增加 TME 中的 M2/M1 巨噬细胞比率 IHC 和流式细胞术在 AIM2 中，我的 K00 解决了更大流行率和潜在的机制。 My K00 将调查 p27pTpT 是否会促进慢性乳腺癌的发生。 通过与雌激素受体（ER）配合激活NFκB/STAT3促进肥胖炎症 免疫逃避和内分泌治疗抵抗我将进一步研究是否与 ER 阻断双重治疗。 检查点抑制剂可以逆转肥胖宿主乳腺癌的内分泌抵抗。 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研究者。