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)： 肝细胞癌和肝内胆管细胞癌通常直到晚期才被发现，5年生存率不到10%。存活率可以， 然而，如果癌症被及早发现，比例将高达40%。不幸的是，通过目前可用的筛查试验，如超声和成像技术，不可能及早发现肝癌/ICC。AFP可作为肝细胞癌的诊断标记物，但其特异性和敏感性较低，限制了其应用。此外，目前还不存在可靠地检测ICC的生物标志物。因此，一种比现有方法更有效地检测肝细胞癌/肝细胞癌的方法将会对治疗提出建议。背景与目的：最近的研究表明，丝氨酸蛋白酶抑制剂Kazal(SPIK1)的水平与肝细胞癌/肝细胞癌的进展密切相关，其过度表达与肝细胞癌/肝细胞癌的短生存期和早期复发密切相关。虽然所有数据表明，SPIK可能被用作检测肝细胞癌/ICC的更有效的生物标志物，但其他疾病如胰腺炎也会导致血清SPIK水平升高。然而，我们已经证明，肝癌分泌的Spik(LC-Spik)与胰腺分泌的Spik有以下区别：(1)LC-Spik的数量明显多于胰腺细胞分泌的量，以及(2)LC-Spik中存在分子结构上的差异，在N端增加了9个氨基酸片段。基于这一发现，我们将能够开发出一种特异性抗体和诊断ELISA检测试剂盒，该试剂盒可以使用LC-Spik作为筛查肝癌/ICC的更有效和更特异的生物标志物。方法：在第一阶段，Aim1：我们将通过构建针对LC-Spik中9个氨基酸片段的特异性单抗来开发一个原型测试系统来定量LC-Spik，该抗体具有高灵敏度(在纳克级)和特异性。AIM2.我们将评估160例肝细胞癌患者和90例肝细胞癌患者的LC-SPIK表现。以正常人、胰腺炎和其他肝病(肝炎和肝硬变)患者的200份血清样本作为对照。在这些研究完成后，我们将验证我们的概念，并获得一个具有高灵敏度和特异性的测试系统，以区分肝细胞癌/肝细胞癌与胰腺炎和其他肝脏疾病或正常肝脏的受试者。在第二阶段研究中，AM3：我们将使我们的技术标准化，并开发一种用于筛查肝细胞癌/肝细胞癌的检测试剂盒(HepatoDetect(R))。该试剂盒可靠、易操作、价格低廉。在Aim4：使用我们的试剂盒，我们将系统地检测来自肝癌、ICC、正常肝脏、胰腺炎以及其他肝脏疾病(如肝炎和肝硬变)的每个临床组的300例样本的LC-SPIK的性能，以确定对肝细胞癌/ICC的特异性。最后，我们将在FDA批准后将该试剂盒商业化。市场：由于美国近1500万人和全球20亿人感染了乙肝病毒或丙型肝炎病毒，其中30-40%的人有患肝癌的高风险。对于这些人来说，常规的癌症检查是必要的。此外，肝细胞癌是仅次于肝癌的第二常见的原发性肝癌，目前尚无可靠的生物标志物用于检测，因此肝细胞癌/肝细胞癌的检测市场是巨大的。竞争：目前，只有AFP被用作肝细胞癌的诊断标记物，但效果较差，市场上还没有类似的产品。 公共卫生相关性： 目前，还没有有效的生物标志物来检测肝癌的早期发生(肝细胞癌和肝细胞癌)及其术后复发。然而，在肝细胞癌和肝细胞癌中发现了丝氨酸蛋白酶抑制因子Kazal(Spik)的高表达，患者血清中的Spik蛋白水平已被证明与肿瘤的进展有关。在这项计划中，我们将开发一种简单快速的诊断试剂盒来特异性地定量血清SPIK，并使用移植了肝癌肿瘤的小鼠的血清样本来检验其有效性，并确定SPIK是否可以作为诊断肝癌/ICC的生物标记物。