Reprogramming the Tumor Microenvironment in Ovarian Cancer

重新编程卵巢癌的肿瘤微环境

基本信息

批准号：
10457939
负责人：
David D Schlaepfer
金额：
$ 40.48万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-03 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10457939
关键词：
Antibodies Antigens Ascites Automobile Driving Bioinformatics CCL2 gene CD8-Positive T-Lymphocytes CD8B1 gene Cancer Etiology Cancer Model Cancer Patient Cause of Death Cells Cessation of life Chemoresistance Chromosomes Combination immunotherapy Complement Disease Endothelial Cells Etiology Exhibits Focal Adhesion Kinase 1 Gene Amplification Genes Genetic Goals Growth ITIM Immune Immune Evasion Immune signaling Immunoassay Immunosuppression Immunotherapy In Vitro Infiltration KRAS2 gene Knock-out Knockout Mice Knowledge Leukocytes Link LoxP-flanked allele Malignant - descriptor Malignant Female Reproductive System Neoplasm Malignant Neoplasms Malignant neoplasm of ovary Mediating Membrane Proteins Messenger RNA Modeling Molecular Molecular Analysis Molecular Conformation Mus Myeloid-derived suppressor cells Nuclear Oncogenes Oncogenic Operative Surgical Procedures Ovarian Ovarian Carcinoma PTK2 gene PTPRC gene Pancreas Pathway interactions Patients Peritoneal Pharmacology Phenotype Phosphotransferases Protein Tyrosine Kinase Resistance Role Serous Signal Transduction Squamous cell carcinoma Supporting Cell System T-Lymphocyte TNF gene TP53 gene Testing Tumor Escape Tumor-Derived Woman cancer recurrence cell motility chemokine chemotherapy combinatorial cyclooxygenase 2 cytokine effector T cell genetic regulatory protein immune checkpoint in vivo inducible gene expression insight kinase inhibitor loss of function mouse model nano-string nectin neoplastic cell ovarian neoplasm poliovirus receptor prevent prognostic significance programmed cell death ligand 1 protein expression receptor expression recruit therapy resistant transcriptome sequencing tumor tumor growth tumor hypoxia tumor microenvironment tumor progression

项目摘要

PROJECT SUMMARY High-grade serous ovarian cancer (HGSOC) is the leading cause of death from gynecologic malignancies. Most patients initially respond to chemotherapy after surgery, yet ~80% will recur with disease that becomes resistant to treatment. Immune therapies have shown great promise, but with limited efficacy in HGSOC. The HGSOC tumor microenvironment (TME) is highly immuno-suppressive and this is hypothesized to promote tumor immune evasion. We have developed two new implantable syngeneic mouse ovarian tumor models that will allow for the molecular analysis of tumor- and host-specific signals driving immune evasion. By selecting for aggressive growth in mice, we have extensively characterized KMF cells (gains in genes for KRas, Myc, and FAK) that exhibit many phenotypic similarities to HGSOC; intrinsic chemo-resistance and potent immune suppression. We will focus on FAK (focal adhesion kinase), a tyrosine kinase canonically supporting cell motility signaling. FAK is the fifth highest amplified gene in HGSOC and greater than 65% of patients exhibit elevated FAK mRNA with poor prognostic significance. Using pharmacological FAK inhibitors, FAK knockout, FAK re-expression, complementation, and bioinformatic analyses of KMF cells in tumor-bearing mice, we find that FAK drives the expression of a select group of cytokines and tumor-associated surface proteins involved in regulating tumor growth and immune evasion. Inhibiting FAK results in decreased myeloid-derived suppressor cell (MDSC) recruitment, increased CD4 and CD8 T cell tumor infiltration, and decreased expression of PD-L1, CD112, and CD155 checkpoint regulatory proteins on KMF cells in vivo. These changes are consistent with a normalization or reprogramming of the ovarian TME by FAK inhibition in a tumor-intrinsic manner. FAK inhibition also prevents bloody ascites formation in the KMF model. A second newly-developed T antigen-driven FAK floxed mouse ovarian carcinoma model (MOVCAR) revealed that FAK loss prevents tumor growth in syngeneic low-T mice. FAK-null MOVCAR tumors were infiltrated by CD45+ leukocytes, and when evaluated in immune-deficient mice, orthotopic FAK-null MOVCAR tumor growth was enhanced. This proposal will test the hypothesis that tumor-intrinsic FAK activation facilitates immune-suppressive related changes to the TME. Aim-1 will use a new inducible FAK expression system to evaluate FAK nuclear localization- and kinase-dependent signals driving malignancy, chemokine expression, and MDSC recruitment. Aim-2 will test the role CD112/CD155 immune checkpoint protein expression and whether FAK inhibition may combine with antibodies to TIGIT (T cell immunoreceptor with Ig and ITIM domains) to limit tumor growth via effects on T cells. Aim-3 will use an inducible knockout of FAK, of the related Pyk2 kinase (new model), or inducible expression of kinase-inactive FAK in mouse endothelial cells, with the KMF implantable tumor model, to test stromal FAK and Pyk2 signaling on the TME. These mouse studies, with analyses of patient tumors, will provide important insights on the role of FAK inhibition to enhance immunotherapy efficacy for HGSOC.

项目摘要高级浆液卵巢癌（HGSOC）是妇科恶性肿瘤死亡的主要原因。大多数患者最初在手术后对化学疗法做出反应，但约有80％会因疾病而复发抗治疗。免疫疗法表现出了很大的希望，但在HGSOC中的疗效有限。这 HGSOC肿瘤微环境（TME）是高度免疫抑制的，这是可以促进的肿瘤免疫逃避。我们已经开发了两个新的可植入式合成小鼠卵巢肿瘤模型将允许对驱动免疫逃避的肿瘤和宿主特异性信号进行分子分析。通过选择对于小鼠的侵略性生长，我们已经广泛地表征了KMF细胞（Kras，Myc，Myc，Myc，Myc的增长 FAK）与HGSOC表现出许多表型相似；固有的化学抗性和有效免疫抑制。我们将专注于FAK（局灶性粘附激酶），酪氨酸激酶在典型的细胞中运动信号传导。 FAK是HGSOC中第五大扩增基因，大于65％的患者表现出升高的FAK mRNA，预后意义较差。使用药理学FAK抑制剂，FAK敲除， FAK重新表达，互补和对肿瘤小鼠中KMF细胞的生物信息分析，我们发现 FAK驱动了一组细胞因子和与肿瘤相关的表面蛋白的表达在调节肿瘤生长和免疫逃避方面。抑制FAK导致髓样衍生的降低抑制细胞（MDSC）募集，CD4和CD8 T细胞肿瘤浸润增加，并减少在体内KMF细胞上PD-L1，CD112和CD155检查点调节蛋白的表达。这些变化与通过肿瘤中的FAK抑制对卵巢TME的归一化或重新编程一致方式。 FAK抑制作用还可以防止KMF模型中的血腥腹水形成。第二个新发达的T 抗原驱动的FAK Floxed小鼠卵巢癌模型（MOVCAR）表明，FAK损失可防止肿瘤合成性低T小鼠的生长。 FAK-NULL MOVCAR肿瘤被CD45+白细胞浸润，当在免疫缺陷型小鼠中评估，原位FAK-NULL MOVCAR肿瘤的生长得到了增强。这个建议将检验以下假设：肿瘤内FAK激活有助于免疫抑制的相关变化 TME。 AIM-1将使用新的诱导FAK表达系统来评估FAK核定位 - 和激酶依赖性信号驱动恶性肿瘤，趋化因子表达和MDSC募集。 AIM-2将测试 CD112/CD155免疫检查点蛋白表达的作用以及FAK抑制是否可能与 TIGIT的抗体（具有Ig和ITIM结构域的T细胞免疫受体），以通过对T的影响限制肿瘤生长细胞。 AIM-3将使用相关PYK2激酶（新模型）或可诱导的FAK的诱导型敲除与KMF植入肿瘤模型的小鼠内皮细胞中激酶无活性FAK的表达 TME上的基质FAK和PYK2信号传导。这些小鼠研究，分析患者肿瘤，将就FAK抑制作用增强HGSOC的免疫疗法功效的作用提供重要的见解。