A novel diagnostic biomarker for hepatocellular carcinoma and cholangiocarcinoma

肝细胞癌和胆管癌的新型诊断生物标志物

• 批准号: 8312069
• 负责人: Xuanyong Lu
• 金额: $ 29.25万
• 依托单位: IMCARE BIOTECH, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8312069
• 关键词: AFP gene; Amino Acids; Antibodies; Apoptosis Inhibitor; Binding; Biological Assay; Biological Markers; Blinded; Cancerous; Cells; Cholangiocarcinoma; Cirrhosis; Clinical; Clinical Research; Clinical Trials; Cultured Cells; Data; Detection; Diagnosis; Diagnostic; Disease; Early Diagnosis; Enzyme-Linked Immunosorbent Assay; Excision; FDA approved; Feasibility Studies; Foundations; Goals; Hepatitis; Hepatitis B Virus; Hepatitis C virus; Hepatocyte; Imaging technology; Implant; Individual; Intrahepatic Cholangiocarcinoma; Liver; Liver Cirrhosis; Liver diseases; Malignant Neoplasms; Malignant neoplasm of liver; Marketing; Medical; Medical Device; Messenger RNA; Methods; Molecular; Monoclonal Antibodies; Mus; Operative Surgical Procedures; Pancreas; Pancreatitis; Patients; Performance; Phase; Primary carcinoma of the liver cells; Procedures; Proteins; Protocols documentation; Reagent; Recurrence; Reference Standards; Sales; Sampling; Screening procedure; Sensitivity and Specificity; Serine Proteinase Inhibitors; Serum; Specificity; Specimen; Staging; Survival Rate; System; Technology; Testing; Transcript; Ultrasonography; Use Effectiveness; base; cancer cell; cancer therapy; high risk; intrahepatic; meetings; milliliter; novel; novel diagnostics; outcome forecast; overexpression; phase 1 study; phase 2 study; prototype; research clinical testing; success; tumor; tumor progression

DESCRIPTION (provided by applicant): Hepatocellular carcinoma (HCC) and intrahepatic cholagiocarcinoma (ICC) usually go undetected until its late stages, with a 5-year survival rate less than 10%. The survival rate can, however, be as high as 40% if cancer is detected early. Unfortunately, early detection of HCC/ICC is not possible by current available screening tests such as ultrasound and image technologies. AFP is used as a diagnostic marker for HCC, however, low specificity and sensitivity limit its use. Additionally, no biomarker currently exists to reliably detect ICC. Therefore, a method of detecting HCC/ICC more effectively than the current methods would bring forward the management. Background and Objective: Recent studies have suggested that levels of serine protease inhibitor Kazal (SPIK) is highly related to HCC/ICC progression and over- expression of SPIK is correlated to short survival and early recurrence of HCC/ICC. While all data suggests that SPIK may be used as a more effective biomarker for detecting HCC/ICC, this had been complicated by other disease such as pancreatitis also led to higher serum SPIK levels. However, we have been able to demonstrate that SPIK secreted by liver cancer (LC-SPIK) can be distinguished from that secreted by pancreas as: (1) the quantity of LC-SPIK is significantly greater than that secreted by pancreatic cells, and (2) a molecular structural difference in the form of an additional 9- amino-acid fragment in the N-terminus is present in the LC-SPIK. Based on this discovery, we would be able to develop a specific antibody and diagnostic ELISA test kit that could use LC-SPIK as a more effective and specific biomarker for screening HCC/ICC than available currently. Approach: In phase I, Aim1: We will develop a prototype test system for quantifying LC-SPIK by constructing a specific monoclonal antibody against the 9 amino-acid segment in LC-SPIK with high sensitivity (at nanogram level) and specificity. Aim2. We will assess the performance of LC-SPIK in 160 patients with HCC and 90 patients with ICC. 200 serum samples from normal people, patients with pancreatitis and other liver disease (hepatitis and cirrhosis) will be used as controls. After completion of these studies we would have proved our concept and obtain a test system with high sensitivity and specificity to distinguish HCC/ICC from the subjects with pancreatitis and other liver diseases or normal liver. In phase II studies, Am3: we will standardize our technology and develop a test kit (HepatoDetect(R)) for screening HCC/ICC. This ELISA kit would be reliable, easy to operate and inexpensive. In Aim4: Using our kit, we will systematically determine the performance of LC-SPIK in a blinded sample sets including 300 specimens in each clinical group from HCC, ICC, normal liver, pancreatitis and other liver diseases such as hepatitis and cirrhosis to determine the specificity for HCC/ICC. Finally we will commercialize this kit after approved by FDA. Market: Because nearly 15 million people in US and 2 billions people in worldwide are infected by HBV or HCV, 30-40% of them have a high risk to develop HCC. For these people, routine examination of cancer is necessary. In addition, ICC is the second most common primary liver cancer after HCC and currently no reliable biomarker exists for detecting it, therefore, the market of detection HCC/ICC is huge. Competition: Currently, only AFP is used as a diagnostic marker for HCC but is less effective, there is no similar product in the market. PUBLIC HEALTH RELEVANCE: Currently, there is no effective biomarker to detect early occurrence of liver cancer (HCC and ICC) and its recurrence after surgical resection. However, overexpression of serine protease inhibitor Kazal (SPIK) was found in HCC and ICC and the level of SPIK protein in the serum of patients has been demonstrated to correlate with the progress of the cancers. In this proposal, we will develop an easy and quick diagnostic kit to specifically quantify serum SPIK and examine its effectiveness using serum samples from mice implanted with HCC tumors and determine whether SPIK can be a biomarker for diagnosis of HCC/ICC.

描述（由申请人提供）： 肝细胞癌（HCC）和肝内胆管癌（ICC）通常一直未被发现到其晚期，5年生存率小于10％。生存率可以， 但是，如果早期检测到癌症，高达40％。不幸的是，通过当前可用的筛选测试（例如超声和图像技术）无法尽早检测HCC/ICC。 AFP用作HCC的诊断标记，但是，特异性低，敏感性限制了其使用。此外，目前尚无生物标志物可靠地检测ICC。因此，一种比当前方法提出管理的方法更有效地检测HCC/ICC的方法。背景和目标：最近的研究表明，丝氨酸蛋白酶抑制剂Kazal（SPIK）的水平与HCC/ICC进展高度相关，而SPIK的过度表达与HCC/ICC的短期生存和早期复发有关。尽管所有数据都表明SPIK可以用作检测HCC/ICC的更有效的生物标志物，但其他疾病（例如胰腺炎）也使这一疾病变得复杂，这也导致血清SPIK水平较高。但是，我们已经能够证明肝癌分泌的SPIK（LC-SPIK）可以与胰腺分泌的SPIK区分开为：（1）LC-SPIK的数量明显大于胰腺细胞分泌的LC-SPIK的数量，并且（ 2）在LC-Spik中存在N末端中额外9-氨基酸片段形式的分子结构差。基于这一发现，我们将能够开发一种特定的抗体和诊断ELISA测试套件，该试剂盒可以将LC-Spik用作比目前可用的更有效，更特定的生物标志物来筛选HCC/ICC。方法：在第一阶段，AIM1：我们将开发一个原型测试系统，用于通过构建针对LC-SPIK中9个氨基酸段的特定单克隆抗体，具有高灵敏度（纳米图水平）和特异性。 AIM2。我们将评估160例HCC患者和90例ICC患者的LC-SPIK的性能。来自正常人，胰腺炎和其他肝病患者（肝炎和肝硬化）的200个血清样本将被用作对照。完成这些研究后，我们将证明我们的概念，并获得具有高灵敏度和特异性的测试系统，以将HCC/ICC与胰腺炎和其他肝病或正常肝脏的受试者区分开。在第二阶段研究中，AM3：我们将标准化我们的技术并开发用于筛查HCC/ICC的测试套件（Hepatodetect（R））。该ELISA套件将是可靠的，易于操作和便宜的。在AIM4中：使用我们的套件，我们将系统地确定在盲人样本组中LC-SPIK的性能HCC/ICC的特异性。最后，我们将在FDA批准后将该套件商业化。市场：由于美国的近1500万人和全世界的20亿人感染了HBV或HCV，因此其中30-40％的人患HCC的风险很高。对于这些人，需要常规检查癌症。此外，ICC是仅次于HCC的第二大原发性肝癌，目前尚无可靠的生物标志物来检测它，因此，检测HCC/ICC的市场很大。竞争：目前，只有AFP用作HCC的诊断标记，但效率较低，市场上没有类似的产品。 公共卫生相关性： 目前，尚无有效的生物标志物来检测肝癌（HCC和ICC）的早期发生及其在手术切除后的复发。但是，在HCC和ICC中发现了丝氨酸蛋白酶抑制剂Kazal（SPIK）的过表达，并且已经证明患者血清中的SPIK蛋白水平与癌症的进展相关。在此提案中，我们将开发一个简单快捷的诊断套件，以专门量化血清SPIK并使用植入HCC肿瘤的小鼠的血清样品来检查其有效性，并确定SPIK是否可以成为诊断HCC/ICC的生物标志物。