Biomarker for Hepatocellular Carcinoma

描述（由申请人提供）：人肝细胞癌（HCC）是慢性肝病和肝硬化最严重的并发症之一。更令人不安的是，从诊断之时起，平均预期寿命约为8至10个月。在美国，HCC的发病率已经开始增加，主要是由于慢性丙型肝炎（HCV）感染的贡献。此外，随着肥胖症的流行，包括II型糖尿病和胰岛素抵抗，非酒精性脂肪性肝炎（NASH）的出现已经开始在这种毁灭性疾病的发病机制中发挥更大的作用，这是由于肝硬化的发展，肝硬化是HCC的危险因素。最近，卫生保健专业人员普遍关注HCC的增加以及缺乏导致或有助于早期诊断和治疗的最佳筛查技术，以期改善这种几乎一致致命的疾病的预后。因此，该建议的目的是验证一种新的标记物，可能有助于HCC的早期诊断。在这方面，我们已经确定了单克隆抗体（mAb）的结合和表达克隆，细胞表面蛋白标记，是在肝癌肿瘤增强。编码这种蛋白质的基因已被克隆并分子鉴定为天冬酰胺基（天冬酰胺基）β-羟化酶（HAAH），其似乎在大多数（>90%）B肝炎（HBV）和HCV相关的HCC中高度表达。此外，这种蛋白质似乎存在于这些患者的血清中。因此，我们已经建立了免疫测定，以检测C和N端片段以及全长HAAH血清中使用重组蛋白作为标准。由于这种酶在细胞表面表达，我们最近开发了针对该分子不同区域的人scFv和IgG抗体。这些试剂将用于生物分布研究，以靶向人HCC小鼠模型中的肿瘤。由于HAAH基因的表达与HCC的发病机制密切相关，因此HAAH基因是HCC的一种新的生物标志物。我们计划采取以下行动。具体目标1。HAAH作为HCC的生物标志物。具体目标2。进一步表征人scFv片段和IgG抗AAH抗体。这些新的基于mAb的测定法可能用于HCC的早期检测，并且人抗HAAH抗体可用作靶向试剂用于肝中HCC的特异性检测。

DESCRIPTION (provided by applicant): Human hepatocellular carcinoma (HCC) is one of the most serious complications of chronic liver disease and cirrhosis. Even more disturbing is the observation that the average life expectancy from the time of diagnosis is approximately eight to ten months. The incidence of HCC has begun to increase in the United States principally due to the contribution of chronic hepatitis C (HCV) infection. In addition, with the obesity epidemic including Type II diabetes and insulin resistance, the emergence of non-alcoholic steatohepatitis (NASH) has begun to assume a larger role in the pathogenesis of this devastating disease due to the development of cirrhosis, a risk factor for HCC. More recently, there is widespread concern among health care professionals regarding the increase in HCC and the lack of optimal screening techniques that lead to or contribute to early diagnosis and treatment in the hope of improving the prognosis of this almost uniformly fatal disease. Thus, the purpose of this proposal is to validate a new marker that may be useful in the early diagnosis of HCC. In this regard, we have identified by monoclonal antibody (mAb) binding and expression cloning, a cell surface protein marker that is enhanced in HCC tumors. The gene encoding for this protein has been cloned and molecularly identified as aspartyl (asparaginyl) (¿-hydroxylase (HAAH) which appears to be highly expressed in most (>90%) hepatitis B (HBV) and HCV related HCC. Furthermore, this protein appears to be present in serum of such patients. Thus, we have established immunoassays to detect C and N terminal fragments as well as full length HAAH in serum using recombinant proteins as standards. Because this enzyme is expressed on the cell surface, we have recently developed human scFv and IgG antibodies directed against the various region of the molecule. These reagents will be used in biodistribution studies to target tumors in murine models of human HCC. Evidence will be presented to support the hypothesis that HAAH gene is a novel biomarker for HCC since expression appears to be highly associated with the pathogenesis of the disease. We plan to do the following. Specific Aim 1. Validate HAAH as a biomarker for HCC. Specific Aim 2. Further characterize human scFv fragments and IgG anti-AAH antibodies. It is likely that these new mAb based assays will be useful for the early detection of HCC and human anti-HAAH antibodies may be employed as targeting reagents for the specific detection of HCC in the liver.