Collaborative Research: Functionalized Nano-textured Surfaces to Isolate and Identify Bladder Cancer Cells

侵袭性和转移性癌症患者的高死亡率在过去几十年中几乎没有改变。 早期发现对于更好地治疗癌症和提高存活率至关重要。 传统的诊断方法无法在非常早期的阶段检测到某些类型的癌症。 膀胱癌通常在早期阶段是无症状的，导致在晚期诊断并且通常是不可治愈的阶段。有时，良性疾病和膀胱癌的症状是非特异性的，非常相似。 在大约85%的癌症患者中，尿液中可能存在红细胞。 但同样的症状可能是尿路（肾脏，膀胱，前列腺）感染或炎症的结果。 该提案旨在提供一种非侵入性，低成本，高灵敏度，和超特异性的方法，使用固态微器件和芯片表面的生物化学功能化。 功能化将靶向在肿瘤细胞上过表达的分子。 将完成数据采集和分析的自动化，以确定肿瘤和正常细胞之间的物理行为差异和生物标志物表达水平。 选择用于靶向检测肿瘤细胞的分子将增加作为治疗结果的指标和预测因子的显著价值。这项提案将改变癌症，特别是膀胱癌的诊断方式。 实验将首先用培养的细胞进行，然后用从携带人膀胱癌异种移植物的动物的血液中分离的细胞进行，最后用真实的患者临床样品进行。多学科团队将整合新技术，为学生提供整个K-Grad连续体的教育机会。研究生和本科生将通过开发细胞-纳米界面交叉列出的课程模块，参与并介绍细胞生物学，生物化学和固态制造的令人兴奋的新维度。研究成果还将用于开发工程学院夏令营的参与性模块，以吸引未来的成年人从事STEM职业。 研究和教育工作的结果、数据和成果不仅将通过同行评审的文章和会议传播，而且还将通过社交和公共媒体传播。该提案的新颖性在于三个要素：首先，将使用一类特殊的探针分子，称为适体。 适体可以可逆地变性，因此这些将提供肿瘤细胞在其天然状态下可逆释放的机制。第二，纳米织构的基底将提供仿生环境以检查细胞行为。第三，肿瘤细胞的动态行为将用于测量单细胞或亚细胞尺度上细胞-细胞相互作用在数天内的时间演变。这种新的肿瘤检测器将用作非侵入性细胞类型报告器（不对细胞进行"染色"），并且作为细胞培养基质，其具有针对一定水平的生物标志物过表达的细胞的规定行为。目前还没有这样的平台。因此，本提案中描述的技术整合可以提供早期识别缓解或转移的手段，甚至可以深入了解给定样本中可能存在多少和什么类型的转移。

The high mortality rate of patients with invasive and metastatic cancer has changed little over the past few decades. Early detection is crucial for better treatment of cancer and to increase survival rates. Traditional diagnostic approaches fail to detect some types of cancers at very early stages. Bladder cancer is usually silent in early stages resulting in diagnosis at late and often incurable stages. Occasionally, the symptoms of both benign conditions and bladder cancer are nonspecific and very similar. In about 85% of cancer patients, there may be red blood cells present in the urine. But the same symptoms may be the result of an infection or inflammation in the urinary tract (kidney, bladder, prostate). This proposal aims at providing a non-invasive, low-cost, highly sensitivity, and ultra-specific approach using solid-state microdevices and biochemical functionalization of chip surfaces. The functionalization will target the molecules that are overexpressed on tumor cells. Automation of the data acquisition and analysis will be done to determine physical behavioral differences and biomarker expression levels between tumor and normal cells. The molecules chosen for targeted detection of tumor cells will add significant value as indicators and predictors of the outcomes of the treatment. This proposal will change how cancers, and especially bladder cancer, are diagnosed. The experiments will be done first with cultured cells, then with cells isolated from the blood of animals bearing human bladder cancer xenografts, and finally with real patient clinical samples. The multi-disciplinary team will integrate the novel technology that will provide educational opportunities for students on the whole continuum of K-Grad. Graduate and undergraduate students will be engaged and introduced to exciting new dimensions of cell biology, biochemistry and solid-state fabrication through development of a cross-listed course module on the cell-nano interface. The research outcomes will be also used to develop participatory modules at College of Engineering summer camps to attract future adults to STEM careers. The results, data and outcomes of the research and education endeavors will be disseminated not only through peer-reviewed articles and conferences, but also through social and public media. The novelty of the proposal lies in three elements: first, a special class of probe molecules called aptamers will be used. Aptamers can be reversibly denatured, and these will therefore provide a mechanism for the reversible release of tumor cells in their native states. Second, the nanotextured substrates will provide biomimetic environment to examine the cell behavior. Third, dynamic behavior of tumor cells will be used to measure the temporal evolution of cell-cell interactions over days at single-cell or sub-cellular scales. This new tumor detector will serve as a non-invasive cell type reporter (without "staining" the cell), and as a cell culture substrate with prescribed behavior of cells for certain level of biomarker overexpression. Currently such a platform does not exist. The technological integration described in this proposal may therefore provide early means of identifying remission or metastases, and even give some insights into how many and what types of metastases may be present in a given sample.