The Potential Role of Firboblast Activation Protein as a Natural Killer Cell Immune Checkpoint in Pancreatic Cancer

成纤维细胞激活蛋白作为自然杀伤细胞免疫检查点在胰腺癌中的潜在作用

基本信息

批准号：
10373963
负责人：
Allison O'Connell
金额：
$ 5.18万
依托单位：
GEORGETOWN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10373963
关键词：
Activated Natural Killer Cell Attenuated Biological Biology Cell Communication Cell Line Cell Surface Receptors Cells Characteristics Cicatrix Clinical Clinical Trials Coculture Techniques Data Desmoplastic Dipeptidyl Peptidases Disease Endopeptidases Exposure to Extracellular Matrix Failure Fibrosis Flow Cytometry Goals Human Immune Immune response Immunosuppression Immunotherapy In Vitro Infiltration Knock-out Knowledge Lesion Life Malignant Neoplasms Malignant neoplasm of pancreas Mediating Modeling Monitor Mus NK Cell Activation Natural Killer Cells Outcome Pancreas Pancreatic Ductal Adenocarcinoma Pattern Pharmacology Phenotype Population Protein Inhibition Public Domains Radiation therapy Receptor Cell Role Sampling Serine Protease Signaling Molecule Survival Rate System Testing Thick Time Tissues Tumor Immunity Work angiogenesis anti-PD1 therapy anti-cancer anti-tumor immune response antitumor effect base cell type chemotherapy clinical application clinically relevant cytokine cytotoxicity fibroblast activation protein alpha fibroblast-activating factor human data imaging system immune cell checkpoints immune clearance immunogenic improved in vitro activity in vivo liquid crystal polymer live cell imaging migration mouse model nano-string novel novel strategies overexpression pancreatic cancer patients pancreatic ductal adenocarcinoma model pancreatic neoplasm pancreatic stellate cell pathology imaging protein activation protein biomarkers protein expression protein function response therapeutic target tool tumor tumor growth tumor-immune system interactions

项目摘要

PROJECT SUMMARY Immunotherapy has been largely ineffective in pancreatic cancer, partially due to the surrounding dense stromal fibrosis which creates an immunosuppressive microenvironment. The main cellular component of this fibrosis, pancreatic stellate cells (PSCs), are marked by elevated expression of fibroblast activation protein (FAP). FAP is a type II transmembrane serine protease that is minimally expressed in normal pancreas, however, in pancreatic cancer FAP is overexpressed in 90% of lesions and is associated with worse clinical outcomes. Here we use a novel in vitro co-culturing system that utilizes primary donor-derived PSCs and a human natural killer (NK) cell line, NK92, to assess the relationship between PSCs and NK cells. We tested the ability of NK cells to kill PSCs and monitored for FAP expression and markers of activation. We also assessed the effect of FAP inhibition on NK cell activity in vitro and pancreatic tumor clearance in vivo. We found that NK cells are activated by and kill PSCs, potentially via NK cell surface receptor NKG2D recognition of MICA/B on PSCs. Upon direct contact with PSCs, PSCs downregulate FAP, however, NK cells upregulate FAP. This is the first-time NK cells have been shown to produce FAP and that induction of FAP is mediated by cell-to-cell contact. Furthermore, FAP expression by NK cells is associated with an inactivate phenotype. FAP inhibition enhanced NK killing of PSCs in vitro and enhanced pancreatic tumor clearance in vivo. The anti-tumor activity of FAP inhibition was enhanced by addition of anti-PD-1 therapy. Based on these findings, I hypothesize that FAP functions as an NK cell immune checkpoint. FAP is expressed in NK cells after activation to attenuate cytotoxicity and can be inhibited to enhance anti-tumor immunity. To test this hypothesis, I aim to determine mechanisms of FAP induction in NK cells (Aim 1), assess the clinical relevance of these findings (Aim 1A and 1D) and manipulate FAP activity to enhance in vitro and in vivo anti-tumor immune responses (Aim 2). Successful completion of these aims will identify factors that regulate FAP expression and further our understanding of how FAP regulates the immune response. These findings will fill the gap in knowledge surrounding regulators of FAP expression and provide new approaches to enhance anti-tumor immune activity.

项目摘要免疫疗法在很大程度上在胰腺癌中无效，部分原因是周围的密集基质纤维化会产生免疫抑制的微环境。这种纤维化的主要细胞成分，胰腺星状细胞（PSC）以成纤维细胞激活蛋白（FAP）的表达升高为标志。 FAP 是一种在正常胰腺中最小表达的II型跨膜丝氨酸蛋白酶胰腺癌FAP在90％的病变中过表达，并且与临床结果较差有关。这里我们使用一种小说的体外共培养系统，该系统利用主要的供体衍生的PSC和人类的自然杀手（NK）细胞系NK92，以评估PSC和NK细胞之间的关系。我们测试了NK细胞的能力杀死PSC并监测以表达FAP和激活标记。我们还评估了FAP的影响体外对NK细胞活性的抑制作用。我们发现NK细胞被激活通过并杀死PSC，可能通过NK细胞表面受体NKG2D识别PSC上的云母。直接与PSC接触，PSC下调FAP，但是，NK细胞上调FAP。这是首次NK细胞已显示出产生FAP，并且FAP的诱导是通过细胞对细胞接触介导的。此外， NK细胞的FAP表达与失活表型有关。 FAP抑制增强了NK杀戮 PSC在体内体外和增强的胰腺肿瘤清除率。 FAP抑制的抗肿瘤活性为通过添加抗PD-1治疗增强。基于这些发现，我假设FAP充当NK 细胞免疫检查点。激活后在NK细胞中表达FAP以减弱细胞毒性，可以是被抑制以增强抗肿瘤免疫力。为了检验这一假设，我旨在确定FAP的机制 NK细胞的诱导（AIM 1），评估这些发现的临床相关性（AIM 1A和1D）并操纵 FAP活性以增强体外和体内抗肿瘤免疫反应（AIM 2）。成功完成这些目标将确定调节FAP表达的因素，并进一步了解FAP如何调节FAP 免疫反应。这些发现将填补围绕FAP表达的调节器的知识的空白并提供新的方法来增强抗肿瘤免疫活性。