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）是女性与癌症相关死亡的主要原因。抗治疗是主要的 限制因素。因此，该赠款的长期目标是阐明新型分子机制 基础疗法抗性。 BC干细胞（CSC）可以通过改变 肿瘤微环境（TME）促进免疫进化。 p27起双重作用：作为肿瘤抑制剂， 抑制细胞周期；作为一种癌基因，当PI3K激活磷酸化时，它会促进转移 T157和T198（p27ptpt）的C末端的激酶。 P27PTPT充当CJUN的转录调节器， 驱动EMT/pro-metanclastic基因谱。我的论文工作表明它也使STAT3共同激活以更新 CSC和癌症促进炎症。我的总体假设是p27ptpt共同激活STAT3 基因谱以扩大癌症干细胞并促进免疫进化。我的论文在aim1.1中的工作 p27ptpt上调的新型观察到包括球体形成和胚胎的干细胞特性 通过共激活STAT3，干细胞转录因子（ES-TFS，MYC，OCT4，NANOG和KLF4）。我的芯片 - SEQ/RNA-SEQ显示了P27招募STAT3到基因启动子，以诱导P27/STAT3共同目标的广泛概况， 包括MYC和JAG1，以增加体内肿瘤发射干细胞（TISC）。我显示了P27PTPT驱动的CSC 扩展部分是通过新型的P27/STAT3共同目标基因Angptl4介导的。在F99中，AIM1.2， 提出以下假设：P27PTPT/STAT3通过重新塑造TME促进肿瘤免疫进化。我会 将我的研究扩展到癌症免疫学上，这为我提供了出色的培训机会。我的 显示P27PTPT的ATAC-SEQ可提高对共核P27/STAT3的染色质的可及性。 致癌基因（IL-6，NF-κB（RERA），VEGFA）和免疫抗性介体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驱动器是否驱动 促进癌症的炎症并将TME塑造为体内肿瘤更为验证状态。首先，我会的 确定P27PTPT如何调节单一调节肿瘤细胞和免疫细胞特征和细胞因子表达 BC的细胞RNA-seq和Luminex在体内肿瘤的外周血中细胞因子的细胞因子测定。我会 还测试P27PTPT如何通过调节CD4，CD8 T细胞和NK细胞浸润来驱动免疫驱逐 TME并测试P27PTPT是否募集MDSC，Tregs并增加TME中M2/M1巨噬细胞的比率 IHC和流式细胞仪。在AIM2中，我的K00解决了较高的患病率和 肥胖女性乳腺癌的死亡率。我的K00会调查P27PTPT是否促进了慢性 通过与雌激素受体（ER）合作与活化的NFB/STAT3合作，肥胖的炎症以促进 免疫进化和内分泌疗法抗性。我将进一步调查是否具有ER封锁双重治疗 检查点抑制剂可以在肥胖宿主的乳腺癌中逆转内分泌耐药性。 F99/K00将 帮助我从肠道前研究到博士后研究，并最终转变为独立的研究者。