I2I phase I: assessment of genotypic markers for profiling of circulating tumor cells

目前的癌症分期方法在应用于预测肿瘤预后和治疗反应时并不广泛有效，这推动了对特定肿瘤特征方法的需求。两个方面混淆了预测肿瘤预后和分配适当治疗的能力，这两个方面是癌症的异质性和难以及时识别转移。一个可能有助于解决这些问题的有希望的方法是对循环肿瘤细胞(CTCs)的分析。由于CTC是从原发肿瘤本身脱落的，这些稀有细胞可以提供一种在治疗过程中重新确定肿瘤特征的方法，而不需要持续的再次活检。这在评估CTC表型以了解疾病侵袭性或在精密医学中应用CTC基因分型时尤其有效。到目前为止，正在开发的四氯化碳技术侧重于捕获和计数这些稀有细胞，并依赖于芯片外分析来评估四氯化碳种群表型和基因类型。因此，不能用单一的解决方案来确定患者血液样本中CTC的特征，目的是开发“液体活组织检查”来为治疗决策提供信息。**为了响应对同时捕获CTC种群并对其进行分子表征的技术需求，我们开发了一种新技术--磁性排名细胞术(MagRC)--它能够在蛋白质和mRNA表达水平上分析CTC。这种方法使用了一种新型的微流控设备，根据结合的纳米颗粒的数量将细胞分类到不同的捕获区，并可以测量根据细胞在设备中的位置计算出的表达水平。拟议的项目旨在严格验证这项技术，评估化验耗材的可制造性，并开发必要的配套技术以实现化验自动化。****

Current cancer staging methods are not broadly effective when applied to predicting tumor prognosis and response to therapy, driving the need for specific tumor characterization approaches. Two aspects which confound the ability to predict tumor prognosis and assign appropriate therapies are the heterogeneous nature of cancers and the difficulty of identifying metastases in a timely manner. A promising approach that may serve to address these problems is the analysis of Circulating Tumour Cells (CTCs). As the CTCs are shed from the primary tumour itself, these rare cells can provide a means to re-characterize a tumour during treatment without the need for continual re-biopsy. This can be particularly powerful when assessing CTC phenotype to understand disease aggressiveness, or CTC genotyping for applications in precision medicine. To this date, technologies being developed for CTCs focus on capture and enumeration of these rare cells and rely on off-chip assays for assessment of CTC population phenotypes and genotypes. Thus, a single solution is not available for characterization of CTCs within a patient's blood sample with the goal of developing a "liquid biopsy" to inform treatment decisions. **In response to a need for technology to simultaneously capture and molecularly characterize a CTC population, we have developed a new technology - Magnetic Ranking Cytometry (MagRC) - that enables CTC profiling at the level of protein and mRNA expression. This approach uses a new type of microfluidic device that sorts the cells into different capture zones according to the number of nanoparticles bound, and can measure expression levels calculated by the location of the cells within the device. The proposed project aims to rigorously validate this technology, assess manufacturability of assay consumables, and develop the necessary companion technologies for assay automation. ****