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

目前的癌症分期方法在预测肿瘤预后和治疗反应方面并不广泛有效，这推动了对特定肿瘤表征方法的需求。癌症的异质性和及时发现转移的困难是影响预测肿瘤预后和分配适当治疗的两个方面。循环肿瘤细胞（ctc）的分析是解决这些问题的一种很有前途的方法。由于ctc从原发肿瘤本身脱落，这些罕见的细胞可以在治疗期间提供一种重新表征肿瘤的方法，而不需要连续的重新活检。当评估CTC表型以了解疾病侵袭性或CTC基因分型用于精准医学应用时，这一点尤其强大。到目前为止，针对CTC开发的技术主要集中在捕获和枚举这些罕见细胞，并依赖于芯片外检测来评估CTC群体表型和基因型。因此，单一的解决方案无法用于患者血液样本中ctc的表征，目的是开发“液体活检”来为治疗决策提供信息。**为了响应同时捕获和分子表征CTC群体的技术需求，我们开发了一种新技术-磁性排列细胞术(MagRC) -能够在蛋白质和mRNA表达水平上分析CTC。这种方法使用一种新型的微流控装置，根据结合的纳米颗粒的数量将细胞分类到不同的捕获区，并且可以通过装置内细胞的位置计算来测量表达水平。拟议的项目旨在严格验证该技术，评估分析耗材的可制造性，并开发分析自动化所需的配套技术。* * * *

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. ****