APPLICATION OF A MULTIPLEXED IMMUNOASSAY FOR THE DETECTION OF BLADDER CANCER

项目摘要/摘要 背景非侵袭性诊断膀胱癌患者以血尿最常见 膀胱癌的临床表现，或有膀胱癌病史的受试者肿瘤监测 仍然是一个挑战，因此，这些患者需要进行侵入性检测。目前以尿液为基础的分析不是 因此不能用来“排除”不需要这些侵入性检测的患者。 此前，我们a)鉴定了一个与膀胱癌相关的诊断蛋白“指纹”，它由10个蛋白质组成。 生物标记物，b)开发了一种多重免疫分析来询问排尿样本中的这10个生物标记物，以及 C)对多重免疫分析进行了分析验证。使用多重免疫分析，我们有 在326名受试者(46例癌症)中产生了令人鼓舞的初步数据，指出敏感性和特异性为93%。 因此，我们第一次拥有了一种准确的检测方法，可以用来排除那些不需要的患者 侵入性测试。此外，早期发现(即在临床表现之前进行检测)是一个重要目标。 适用于有患膀胱癌风险的患者。在报告中，超过70%的膀胱癌病例是非 肌肉浸润性膀胱癌(NMIBC)，其余30%为肌肉浸润性膀胱癌 (MIBC)或转移性。早期发现(即NMIBC)，5年存活率约为94%， 相比之下，当该疾病被检测为肌肉浸润性膀胱癌时，5年生存率为~50% (MIBC)，当疾病是转移性时&lt；20%。然而，到目前为止，还没有可用的早期检测方法。在一个 来自我们正在进行的R01研究的20名膀胱癌患者和20名匹配对照的小队列，我们有 早在18个月前就注意到与膀胱癌相关的诊断蛋白“指纹”的增加。 对膀胱癌的临床诊断和对所有膀胱癌的实际阳性多重免疫检测 患者在临床诊断膀胱癌前12个月就诊。假设：与膀胱癌相关的疾病 诊断蛋白质“指纹”的存在可以被用来指示非膀胱癌患者的存在。 不仅在诊断时，而且在临床表现和 膀胱癌的诊断。具体目的：1)验证膀胱癌多重免疫检测方法的有效性 在一项大型嵌套病例对照研究中的检测(n=800)和2)评价早期多重免疫分析 一项大型嵌套病例对照研究中的检测(n=600)。这项研究的意义将为 改进非侵入性检测膀胱癌的方法，将有显著的意义 对病人护理的影响。方法论之前，我们小组已经发现并进行了早期验证 与膀胱癌相关的诊断蛋白质“指纹”，结果非常令人鼓舞。在当前 建议，我们现在寻求在两个大型嵌套病例对照研究中测试多重免疫分析，这两个研究 将允许我们开发和锁定体外诊断多变量指数分析(IVDMIA)算法 AIMS中列出了两个明显的迹象。这样的算法可以在未来的前瞻性研究中部署。 预期结果存在尚未满足的临床需求，需要可靠的生物标记物a)“排除”哪些患者 有血尿或对膀胱癌的监测不需要进一步评估和b)早期发现 当膀胱癌更容易治疗时，存活率也会提高。实现这样一种稳健的分析方法 可能会产生深远的影响，从而改善医疗保健并降低医疗成本。

Project Summary/Abstract Background The non-invasive diagnosing of bladder cancer in patients with hematuria, the most common clinical presentation of bladder cancer, or in subjects with a history of bladder cancer on tumor surveillance remains a challenge, and as such, these patients require invasive testing. Current urine-based assays are not robust and therefore cannot be used to ‘rule out’ patients who do not require these invasive testing. Previously, we a) identified a bladder cancer-associated diagnostic protein “fingerprint” comprised of 10 biomarkers, b) developed a multiplex immunoassay to query these 10 biomarkers in voided urine samples and c) performed analytical validation of the multiplex immunoassay. Using the multiplex immunoassay, we have generated encouraging preliminary data in 326 subjects (46 cancer) noting a sensitivity and specificity of 93%. Thus, for the first time, we possess an accurate assay that can be used to ‘rule out’ patients who do not require invasive testing. Furthermore, early detection (i.e., detection prior to clinical manifestation) is an important goal for patients at risk for bladder cancer. At presentation, more than 70% of bladder cancer cases are non- muscle invasive bladder cancer (NMIBC), whilst the remaining 30% are muscle invasive bladder cancer (MIBC) or metastatic. When detected early (i.e., NMIBC), the 5-year survival rate is approximately 94%, compared to a 5-year survival rate of ~50% when the disease is detected as muscle invasive bladder cancer (MIBC), and <20% when the disease is metastatic. However, to date, no early detection assay is available. In a small cohort of 20 bladder cancer patients and 20 matched controls from our ongoing R01 studies, we have noted an elevation in our bladder cancer-associated diagnostic protein “fingerprint” as early as 18 months prior to the clinical diagnosis of bladder cancer and an actual positive multiplex immunoassay in all bladder cancer patients 12 months prior to the clinical diagnosis of bladder cancer. Hypothesis: A bladder cancer-associated diagnostic protein “fingerprint” exists that can be leveraged to indicate the presence of bladder cancer in non- invasively obtained urine samples not only at the time of diagnosis, but prior to the clinical presentation and diagnosis of bladder cancer. Specific Aims: 1) To validate the multiplex immunoassay for bladder cancer detection in a large, nested case-control study (n=800) and 2) To evaluate the multiplex immunoassay for early detection in a large, nested case-control study (n=600). Significance This research will open the door for improving on the non-invasive methods for detecting bladder cancer and as such it will have a marked impact on patient care. Methodology Previously, our group has discovered and performed early validation of bladder cancer-associated diagnostic protein” fingerprint’ with extremely encouraging results. In the current proposal, we now seek to test the multiplex immunoassay in two large, nested case control studies, which would allow us to develop and lockdown In Vitro Diagnostic Multivariate Index Assay (IVDMIA) algorithms for two distinct indications listed in the aims. Such algorithms can be deployed in future prospective studies. Expected Results There exists an unmet clinical need for reliable biomarkers a) to ‘rule out’ which patients with hematuria or on bladder cancer surveillance do not require further evaluation and b) to early detect bladder cancer when its more treatable with improved survival rates. Implementation of such a robust assay could have a profound impact leading to improved care and reduced healthcare costs.