A microRNA based prognostic service that predicts melanoma metastasis likelihood

摘要 黑色素瘤是一种高度流行的癌症，通过转移和破坏远端器官杀死患者。到 迄今为止，没有有效的分子预后服务可用于对高风险患者进行分层 用于辅助治疗中的转移性复发。通过将患者分为高风险和低风险， 可以为那些能够从治疗中受益的患者量身定制，同时保留那些不会受益的患者 复发，不需要昂贵和潜在有害的治疗。最近的研究发现了三种miRNAs （miR-1908、miR-199a5p和miR-199a3p），其在转移性黑素瘤细胞中过表达并抑制 载脂蛋白A（ApoE）的表达，一种转移抑制因子在黑色素瘤中。癌细胞分泌的ApoE 通过与黑色素瘤细胞LRP 1结合防止细胞侵袭和转移性内皮募集， 内皮细胞LRP 8受体。重要的是，在原发性肝癌中， 肿瘤样本在将黑色素瘤患者分为高可能性的患者和低可能性的患者方面表现稳健。 转移复发的可能性很低在这里，Rgenix建议开发一种预测服务， 预测黑色素瘤转移复发的可能性，基于基因表达分析， 原发性肿瘤载玻片中的miRNA/ApoE基因签名。长期目标是提供一种服务， 全世界诊断为黑色素瘤的患者进行筛查，并分析其发展风险 转移这些信息将指导临床医生做出关键决策，以准确治疗每种疾病。 病人为此，我们建立了一种通过实时荧光定量分析这三种miRNA的联合表达的方法。 将开发PCR和ApoE免疫组化，以将患者分为高风险和低风险患者 阳性预测值> 80%。此外，Rgenix将确定灵敏度 和特异性，并为每种miRNA和ApoE的强度建立最佳阈值 这将允许对高风险与低风险复发者进行分层。A> 80%灵敏度， 特异性将证明该方法的可行性，并允许进行第二阶段。在第二阶段， I期确定的生物标志物组合将在大型回顾性患者队列中进行验证， 提高，以达到> 85%的特异性和灵敏度。该阈值将用于启动前瞻性 与Memorial Sloan Kettering癌症中心合作进行的临床试验。此外，CLIA-CAP认证实验室 将建立和Rgenix将促进产品，并获得第三方报销。Rgenix预后 服务将被证明是无价的社会，这是经历越来越多的黑色素瘤发病率， 临床社区，其受到经济力量的限制，以限制转移预防治疗， 转移复发风险最高的患者。

ABSTRACT Melanoma is a highly prevalent cancer that kills patients by metastasizing and destroying distal organs. To date, there are no effective molecular prognostic services available for stratifying patients that are at high risk for metastatic relapse in the adjuvant setting. By stratifying patients into high-risk versus low-risk, therapies could be tailored to those patients that stand to benefit from treatment while sparing those patients who will not relapse and not require costly and potentially harmful therapies. Recent research identified three miRNAs (miR-1908, miR-199a5p, and miR-199a3p) that are overexpressed in metastatic melanoma cells and inhibit expression of apolipoprotein A (ApoE), a metastasis suppressor in melanoma. Cancer-cell secreted ApoE prevents cellular invasion and metastatic endothelial recruitment through engaging melanoma cell LRP1 and endothelial cell LRP8 receptors, respectively. Importantly, elevated expression levels of miRNAs in primary tumor samples performs robustly in stratifying melanoma patients into those with high likelihood from those with very low likelihood for metastatic relapse. Here, Rgenix proposes to develop a prognostic service that predicts the likelihood of melanoma metastasis relapse, based on gene expression analysis of the miRNA/ApoE gene signature in primary tumor slides. The long-term goal is to provide a service that allows all patients diagnosed worldwide with melanoma to be screened and profiled for their risk of developing metastasis. This information will guide clinicians in the critical decision-making for accurate treatment of each patient. To that aim, a protocol for combined expression analysis of the three miRNA by quantitative real-time PCR and ApoE by immunohistochemistry will be developed to stratify patients into those at high versus low risk for metastasis relapse with a >80% positive predictive value. Furthermore, Rgenix will determine the sensitivity and specificity of the assay and establish an optimal threshold for the intensity of each miRNA and ApoE expression that will allow for stratification of high-risk versus low-risk relapsers. A >80% sensitivity and specificity will proof feasibility of the approach and allow proceeding to Phase II. In Phase II the threshold and biomarker combination determined in Phase I will be validated in a large retrospective patient cohort, and improved in order to reach >85% specificity and sensitivity. This threshold will be used to initiate prospective trials in collaboration with Memorial Sloan Kettering Cancer Center. In addition, a CLIA-CAP certified laboratory will be set up and Rgenix will promote the product, and obtain 3rd party reimbursement. Rgenix' prognostic service would prove invaluable for society, which is experiencing increasing incidence of melanoma, and for the clinical community, which is constrained by economic forces to restrict metastasis prevention therapies to patients at highest risk for metastatic relapse.