Non-invasive Differentiation of Benign Lesions from Aggressive Pancreatic Cancer

良性病变与侵袭性胰腺癌的无创鉴别

• 批准号: 8823694
• 负责人: Charles Scott Craik
• 金额: $ 17.21万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-01-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8823694
• 关键词: Address; Anatomy; Animal Model; Benign; Binding; Biochemical; Biodistribution; Biological Assay; Cancer cell line; Cathepsins; Cells; Cellular Membrane; Characteristics; Cleaved cell; Clinic; Clinical; Data; Detection; Disease; Dissection; Doctor of Medicine; Early Diagnosis; Evaluation; Excision; Exhibits; Figs - dietary; Genetically Engineered Mouse; Goals; Harvest; Human; Image; Imaging Techniques; Immunofluorescence Immunologic; Intraepithelial Neoplasia; Investigation; Knockout Mice; Label; Lasers; Lead; Lesion; Link; Malignant Neoplasms; Malignant neoplasm of pancreas; Masks; Mass Spectrum Analysis; Membrane; Methods; Modeling; Mucinous Neoplasm; Mus; Noise; Nuclear; Operative Surgical Procedures; Pancreas; Pancreatic Diseases; Pancreatic Ductal Adenocarcinoma; Papillary; Pathology; Patients; Peptide Hydrolases; Peptides; Pharmacodynamics; Positron-Emission Tomography; Proteolysis; Radioisotopes; Radionuclide Imaging; Risk; Signal Transduction; Stratification; Substrate Specificity; Surface; Surgeon; Testing; Tissues; Transgenic Mice; Transgenic Model; Translations; Work; acute pancreatitis; antimicrobial peptide; base; cancer cell; chronic pancreatitis; design; fluorescence imaging; imaging agent; imaging probe; improved; in vivo; laser capture microdissection; mouse model; new technology; non-invasive imaging; optical imaging; pancreatic cancer cells; pancreatic neoplasm; prevent; protein aminoacid sequence; prototype; public health relevance; tumor; tumor progression

DESCRIPTION (provided by applicant): At present, we cannot differentiate benign lesions of the pancreas from aggressive tumors without surgical resection and subsequent histological evaluation. The long-term goal of this proposal is to develop a positron emission tomography (PET) imaging agent for clinical use to improve risk stratification among pancreatic neoplasms. The goal of this R21 project is to establish proof of feasibility by demonstrating successful imaging of pre-invasive and invasive tumors in an animal model using our protease activity probes with optical and radionuclide imaging techniques. Recent work has shown cathepsin-like protease activity correlates with pancreatic intraepithelial neoplasm (PanIN) grade. These data raise the possibility that such activity could also be used to differentiate benign lesions from aggressive cancer. We have developed new technology that sensitively and non-invasively detects protease activities. Preliminary studies have shown promise for non-invasively detecting, differentiating, and grading pancreatic diseases including acute and chronic pancreatitis, intraductal papillary mucinous neoplasm (IPMN), PanIN, and pancreatic ductal adenocarcinoma (PDAC). These data support pursuing the following aims to test our hypothesis that we can use proteolysis to activate a membrane binding imaging agent for accurate and quantitative non-invasive imaging of pancreatic cancer to provide functional and dynamic information about tumor status. By using optical imaging to optimize the probe and radionuclide imaging we intend to show proof of principle for ultimately developing a PET imaging agent for clinical use. Aim 1. Optimize and Validate Lead Agents for the Detection of Pancreatic Cancer. The probes will be optimized to increase selectivity and signal to noise and to verify that they work in a variety of cancer cell lines. Probes will be ranked by fluorescence imaging assays in panels of mouse and human pancreatic cancer cell lines. Aim 2. Optimize Probes using Non-Invasive Imaging of Pancreatic Cancer Progression in a Transgenic Model of Pancreatic Cancer. The best probes from Aim 1 will be evaluated in mice. We will use a genetically-engineered mouse model of pancreatic cancer where pancreas-directed expression of constitutively active Kras drives PanIN formation. Probe accumulation, as detected with optical imaging and in sectioned tissue will be correlated to histological grade and oriented by immunofluorescence to cell specific markers. Laser capture microdissection will be used to harvest regions of high proteolytic activity and determine which protease(s) are present with MS. These data will directly link specific protease activities to pancreatic cancer and their precursor lesions in vivo. Based on results from optical imaging, a small number of selected probes will be labeled with radionuclides and PET imaging will be conducted. Pharmacodynamics and biodistribution studies will be carried out to demonstrate translatability to the clinic. These aims will determine whether pericellular proteolysis can be used to identify pancreatic disease and whether different pathologies can be distinguished non-invasively.

描述（申请人提供）：目前，我们无法区分胰腺良性病变与侵袭性肿瘤，除非手术切除并进行组织学评估。本提案的长期目标是开发一种用于临床的正电子发射断层扫描（PET）显像剂，以改善胰腺肿瘤的风险分层。这个R21项目的目标是通过展示我们的蛋白酶活性探针与光学和放射性核素成像技术在动物模型中成功成像侵袭前和侵袭性肿瘤，从而建立可行性证明。最近的研究表明，组织蛋白酶样蛋白酶活性与胰腺上皮内肿瘤（PanIN）分级相关。这些数据提高了这种活性也可用于区分良性病变和侵袭性癌症的可能性。我们开发了一种新技术，可以灵敏地、无创地检测蛋白酶的活性。初步研究显示，该技术有望用于无创检测、鉴别和分级胰腺疾病，包括急性和慢性胰腺炎、导管内乳头状粘液瘤（IPMN）、PanIN和胰腺导管腺癌（PDAC）。这些数据支持我们追求以下目标，以验证我们的假设，即我们可以使用蛋白水解激活膜结合显像剂，对胰腺癌进行准确定量的无创成像，以提供肿瘤状态的功能和动态信息。通过使用光学成像来优化探针和放射性核素成像，我们打算证明最终开发用于临床使用的PET显像剂的原理。目的1。优化和验证胰腺癌先导药物的检测。这些探针将被优化，以提高选择性和信号噪声，并验证它们在各种癌细胞系中起作用。探针将在小鼠和人类胰腺癌细胞系面板上进行荧光成像测定。目标2。在胰腺癌转基因模型中使用无创成像技术优化胰腺癌进展探针。来自Aim 1的最佳探针将在小鼠中进行评估。我们将使用基因工程小鼠胰腺癌模型，其中胰腺定向表达组成活性Kras驱动PanIN的形成。通过光学成像和切片组织检测到的探针积累将与组织学分级相关，并通过免疫荧光定向到细胞特异性标记物。激光捕获显微解剖将用于获取高蛋白水解活性区域，并确定ms中存在哪些蛋白酶。这些数据将直接将特定蛋白酶活性与胰腺癌及其体内前体病变联系起来。根据光学成像的结果，少量选定的探针将被放射性核素标记，并进行PET成像。药效学和生物分布研究将进行，以证明转化到临床。这些目的将决定细胞外蛋白水解是否可以用于识别胰腺疾病，以及是否可以无创地区分不同的病理。