Glycoproteins and Glycan-Binding IgGs: Biomarkers for Cancer and Inflammatory Di

描述（由申请方提供）：结构不同的聚糖对炎症和癌症进展显示出出乎意料的强作用。聚糖结合蛋白对聚糖的识别与生理和疾病相关的细胞粘附或细胞信号传导有关。肿瘤组织分泌糖蛋白，其诱导自身抗体产生。癌症中的糖蛋白聚糖水平和结构不同于良性或炎性疾病中发现的那些。最近，一些癌症特异性聚糖生物标志物被确定通过筛选微阵列聚糖与免疫球蛋白（IG）馏分分离自乳腺癌和经典霍奇金淋巴瘤（cHL）患者。对于这两项研究，随后采用较大样本量的直接ELISA验证了聚糖生物标志物。这些研究表明，免疫球蛋白识别的聚糖是疾病特异性和可靠的生物标志物。前列腺特异性抗原（PSA）核心蛋白水平的增加已被广泛用于检测，分期和监测前列腺癌。然而，PSA水平有时不能区分癌症与良性或炎性疾病。在最近的一项研究中，来自无癌症个体的250份血浆样本中约有20份的PSA水平高于4 ng/ml（PSA假阳性），可能是由于BPH或前列腺炎。此外，PSA水平在4 - 10 ng/ml之间的个体的活检样本中约75%为肿瘤阴性。因此，我们建议确定前列腺癌的生物标志物，以弥补PSA测试的特异性的缺点。高通量技术（糖蛋白和聚糖微阵列）和分离技术的组合依赖于聚糖结构和IgG或IgM（与癌症有关的2种主要Ig）的聚糖结合特异性，显示出获得区分前列腺癌和BPH的分子特征的希望。在I期，我们将（a）通过凝集素/抗体微阵列分析用26种糖蛋白生物标志物候选物鉴定前列腺癌特异性血浆糖蛋白（核心蛋白和聚糖）生物标志物，例如，糖基化PSA，（B）使用选自9种不同长度和疏水性的内部合成接头的聚糖接头开发前列腺癌特异性70聚糖微阵列，和（c）检测IgG或IgM的聚糖结合谱，以通过聚糖微阵列获得前列腺癌特异性生物标志物。将使用统计学和生物信息学工具鉴定糖蛋白或聚糖IG对的单个或特征性前列腺癌生物标志物。将使用假PSA阳性血浆样本进一步研究生物标志物的特异性。在II期期间，将基于I期研究产生用于广泛血浆和组织筛查的临床直接ELISA和聚糖子阵列。将研究前列腺癌风险和侵袭性增加与寡糖生物合成途径多态性的关系。在第一阶段和第二阶段开发的新概念和技术可以应用于其他癌症或炎症性疾病的生物标志物的一般搜索与各种生物液体和组织。对自身抗体识别的前列腺癌特异性聚糖的鉴定将导致开发基于聚糖的治疗剂。 公共卫生相关性：由于美国预期寿命的增加，患有慢性炎症性疾病和癌症的人口比例每年都会增加。癌症生物标志物有时无法区分早期癌症与良性或炎性疾病。前列腺癌是美国男性最常见的恶性肿瘤。目前，血清PSA的测量用于癌症诊断，但PSA测定的特异性不令人满意。PSA水平在4 - 10 ng/ml之间的个体的活检样本中，约75%为肿瘤阴性。因此，我们建议寻找寡糖相关的前列腺癌生物标志物，用于早期前列腺癌诊断，以弥补PSA检测特异性的不足。为前列腺癌诊断开发的新概念和技术可用于检测其他癌症和炎性疾病。

DESCRIPTION (provided by applicant): Structurally varied glycans displayed unexpectedly strong efects on inflammation and cancer progression. Glycan recognition by glycan-binding proteins is involved with physiological and disease-related cell adhesion or cell signaling. Tumor tissues secrete glycoproteins, which induce autoantibody production. Glycoprotein glycan levels and structures in cancer differ from those found in benign or inflammatory diseases. Recently, a few cancer-specific glycan biomarkers were identified by screening microarrayed glycans with immunoglobulin (Ig) fractions isolated from breast cancer and classical Hodgkin's lymphoma (cHL) patients. For both studies, subsequent direct ELISAs with larger sample sizes verified the glycan biomarkers. These studies demonstrated that glycans recognized by Igs are disease-specific and reliable biomarkers. Increased prostate-specific antigen (PSA) coreprotein levels have been widely used to detect, stage and monitor prostate cancer. However, PSA level sometimes cannot distinguish cancer from benign or inflammatory diseases. In a recent study, ~20 out of 250 plasma samples from cancer-free individuals had PSA levels higher than 4 ng/ml (false PSA positives) probably due to BPH or prostatitis. Moreover, ~75% of biopsy samples of individuals with PSA levels between 4 - 10 ng/ml were tumor negative. Thus, we propose to identify prostate cancer biomarkers to compensate for shortcomings in specificity of the PSA test. Combination of high-throughput technologies (glycoprotein and glycan microarrays) and fractionation technologies dependent on glycan structures and glycan-binding specificity of the IgG or IgM (2 major Igs involved with cancer) show promise of obtaining molecular signatures to distinguish prostate cancer from BPH. During Phase I, we will (a) identify prostate cancer-specific plasma glycoprotein (coreprotein and glycan) biomarkers by lectin/antibody microarray analyses with 26 glycoprotein biomarker candidates, e.g., glycosylated PSA, (b) develop prostate cancer-specific 70 glycan microarrays using glycan linkers selected among 9 in-house synthesized linkers of various lengths and hydrophobicity and (c) detect glycan-binding profiles of IgGs or IgMs to obtain prostate cancer-specific biomakers by the glycan microarray. Single or signature prostate cancer biomarkers of glycoprotein or glycan Ig pair will be identified using Statistics and Bioinformatics tools. Specificity of the biomarkers will be further investigated wit the false PSA positive plasma samples. During Phase II, clinical direct ELISAs and glycan subarrays for extensive plasma and tissue screening will be produced based on the Phase I study. Increased prostate cancer risk and aggressiveness will be studied in relation to oligosacharide biosynthesis pathway polymorphisms. New concepts and technologies developed during Phases I and II can be applied for general searches of biomarkers for other cancers or inflammatory diseases with various biological fluids and tissues. Identification of prostate cancer- specific glycans recognized by autoantibodies will lead to development of glycan-based therapeutics. PUBLIC HEALTH RELEVANCE: Due to increased life expectancy in the US, the percentage of the population suffering from chronic inflammatory diseases and cancers will increase each year. Cancer biomarkers sometimes fail to distinguish early stage cancer from benign or inflammatory diseases. Prostate cancer is the most common malignancy in American men. Currently, the measurement of serum PSA is used for cancer diagnosis but specificity of the PSA assay is not satisfactory. Approximately 75% of biopsy samples of individuals with PSA levels between 4 and 10 ng/ml were tumor negative. Thus, we propose to search oligosaccharide-related prostate cancer biomarkers for early prostate cancer diagnosis to compensate for shortcomings in specificity of the PSA test. New concepts and technologies developed for the prostate cancer diagnostics can be utilized for detection of other cancers and inflammatory diseases.