A Multiplex Protein Biomarker-Based Immunoassay for the Early Detection of Bladder Cancer and its Implications in Tumor Biology

项目总结/摘要 到2021年，超过83，000名美国人将被诊断患有膀胱癌，超过17，000人死于膀胱癌。 这一时期的疾病。不幸的是，膀胱癌的绝对病例数和死亡人数 自2000年以来，癌症分别增加了57%和41%。当早期检测到（即，NMIBC或阶段1）， 5年生存率> 90%，相比之下，如果疾病被注意到是MIBC，则生存率显著降低 （2期; 50%5年存活率）或转移性（3期和4期; <20%5年存活率）。因此，普遍的看法是， 在高危个体中膀胱癌的早期检测（即，暴露于某些致癌物质的个体）将 可能是解决晚期膀胱癌悲惨结局的最佳方式。目前，对 处于风险中的个人仍然是一个挑战，因此，没有任何方法可以有效地筛查这一点。 高危人群。先前，我们已经a）确定了膀胱癌相关的诊断“特征” 由10种生物标志物组成，B）开发了多重免疫测定以查询排泄尿中的“特征 样品和c）进行多重免疫测定的分析验证。使用多重免疫测定， 我们已经从362名受试者（46种癌症）的队列中获得了令人鼓舞的初步数据（AUC 0.95; 敏感性0.93，特异性0.93，阳性预测值0.65，阴性预测值0.99）。因此，对于 第一次，我们拥有了一个强大的分析，可用于非侵入性检测膀胱癌。利用该 在我们正在进行的前瞻性研究中，我们对有膀胱癌病史的患者进行了监测，我们注意到 早在癌症临床诊断前18个月，我们的“签名”就已升高， 在癌症临床诊断前12个月，在所有癌症患者中进行多重免疫测定。此外，委员会认为， 我们有证据表明，我们的10个生物标志物中的9个在“签名”中表达于相关的致癌物诱导的癌症中。 小鼠膀胱癌模型。假设：存在膀胱癌相关特征，可以利用 以在临床检查前数月至数年从单个排泄尿液样品指示膀胱癌的存在。 膀胱癌的临床表现和诊断。具体目标：1）进行试点研究，以评估 多重免疫测定早期检测膀胱癌的能力和2）使用相关致癌物诱导 小鼠膀胱癌模型，以确定肿瘤微环境内的早期变化， 用于在人类样品中测试的生物标志物候选物。这项研究将打开大门， 改善膀胱癌的早期检测的非侵入性方法，因此，它将具有 对患者生存率有显著影响。方法我们将进行一项前瞻性试点研究（n=150）， 证明了识别、跟踪和检测膀胱癌高危人群的可行性。然后 利用相关致癌物诱导的小鼠膀胱癌模型，我们将研究空间和时间 关联我们的“签名”，并将其与肿瘤微环境中的关键变化联系起来， 生物标志物用于未来的临床开发。预期结果存在未满足的临床需求， 早期发现膀胱癌的生物标志物，当它更可治疗时，提高生存率。

Project Summary/Abstract Background Over 83,000 Americans will be diagnosed with bladder cancer in 2021 with over 17,000 dying of the disease during this period. Unfortunately, both the absolute numbers of cases and deaths from bladder cancer have increased by 57 and 41%, respectively, since 2000. When detected early (i.e., NMIBC or stage 1), the 5-yr survival rate is >90%, compared to a significant reduction in survival if the disease is noted to be MIBC (stage 2; 50% 5-yr survival) or metastatic (stages 3 and 4; <20% 5-yr survival). Thus, the prevailing idea is that early detection of bladder cancer in high risk individuals (i.e., individuals exposed to certain carcinogens) will likely be the best modality to address advanced bladder cancer’s dismal outcomes. Currently, the evaluation of at risk individuals remains a challenge, and as such, there are no modalities available to effectively screen this high risk population. Previously, we have a) identified a bladder cancer-associated diagnostic “signature” comprised of 10 biomarkers, b) developed a multiplex immunoassay to query the “signature” in voided urine samples and c) performed analytical validation of the multiplex immunoassay. Using the multiplex immunoassay, we have generated encouraging preliminary data from a cohort of 362 subjects (46 cancers) (AUC 0.95; sensitivity 0.93, specificity 0.93, positive predictive value 0.65 and negative predictive value 0.99). Thus, for the first time, we possess a robust assay that can be used to non-invasively detect bladder cancer. Utilizing this assay in our ongoing prospective study surveilling patients with a history of bladder cancer, we have noted an elevation of our ‘signature’ as early as 18 months prior to the clinical diagnosis of cancer and an actual positive multiplex immunoassay in all cancer patients 12 months prior to the clinical diagnosis of cancer. Furthermore, we have evidence that 9 of our 10 biomarkers within the “signature” are expressed in relevant carcinogen induced mouse bladder cancer model. Hypothesis: A bladder cancer-associated signature exists that can be leveraged to indicate the presence of bladder cancer from a single voided urine sample months to years prior to the clinical presentation and diagnosis of bladder cancer. Specific Aims: 1) To perform a pilot study to evaluate the multiplex immunoassay’s ability to early detect bladder cancer and 2) To use a relevant carcinogen induced mouse bladder cancer model to identify early changes within the tumor microenvironment which could serve as biomarkers candidates for testing in human samples. Significance This research will open the door for improving on the non-invasive methods for the early detection of bladder cancer, and as such, it will have a marked impact on patient survival. Methodology We will conduct a prospective pilot study (n=150) to demonstrate the feasibility of identifying, following and testing high risk individuals for bladder cancer. Then utilizing a relevant carcinogen induced mouse bladder cancer model, we will study the spatial and temporal association of our “signature” and link it to key changes within the tumor microenvironment, identifying novel biomarkers for future clinical development. Expected Results There exists an unmet clinical need for reliable biomarkers to early detect bladder cancer when its more treatable with improved survival rates.