MicroSNARE: Automated Enrichment Of Circulating Tumour DNA For Improved Cancer Treatment

目前，为了选择最好的治疗方案，大多数癌症患者需要进行一次不愉快且昂贵的外科手术，从肿瘤中取出组织样本，即活组织检查。然而，所有人的血液中都有少量自由循环的DNA（不在他们的血细胞中），而在癌症患者中，一些循环的自由DNA来自他们的肿瘤。这种肿瘤DNA与健康的循环DNA不同，因为它携带了将这些细胞从健康细胞转变为癌细胞的突变。因此，这种循环肿瘤DNA （ctDNA）有可能揭示患者癌症的细节，并显示哪种治疗方法最有可能成功。研究ctDNA的方法已经开发出来，但都受到血液样本中ctDNA丰度低和存在大量背景健康DNA的限制。我们的建议是开发处理血液样本的方法，降解健康的DNA序列，并通过丰富信息丰富的肿瘤DNA，使其更容易检测和表征。我们开发了一种新方法（SNARE），我们希望对现有的有前途的富集方法（NaME-PrO）进行测试。这些方法的广泛应用将使患者受益，需要它们的自动化，我们将在机器人台式和微流体平台上开发。最后，我们将使用一组乳腺癌患者的血液样本定量评估最佳方法，这些方法应该能够更灵敏地检测这些样本中的ctDNA，并且需要更便宜的DNA测序。总之，该项目应该有助于使癌症患者血液样本的成功分析更便宜、更可靠，从而减少所需的活检手术次数，并使更多的循证癌症治疗成为可能。

Currently, in order to select their best treatment options, most cancer patients need to have an unpleasant and expensive surgical operation to remove a sample of tissue from their tumour, called a biopsy. However, all people have small amounts of DNA circulating freely in their blood (which is not inside their blood cells) and in cancer patients some of this circulating free DNA has come from their tumour. This tumour DNA is different from their healthy circulating DNA as it carries the mutations which have transformed these cells from healthy cells to cancer cells. Therefore this circulating tumour DNA (ctDNA) has the potential to reveal details about the patient's cancer and show which treatments are most likely to be successful. Methods to study ctDNA have been developed but all are limited by the low abundance of ctDNA and the presence of larger amounts of background healthy DNA in blood samples. Our proposal is to develop methods to process blood samples, degrading healthy DNA sequences and by enriching the informative tumour DNA, making it easier to detect and characterise. We have developed a new method (SNARE) which we wish to test against an existing promising enrichment method (NaME-PrO). The widespread use of these methods to benefit patients will require their automation, which we will develop in both robotic benchtop and microfluidic platforms. Finally, we will quantitatively assess the best performing methods using a set of blood samples from breast cancer patients, which should give more sensitive detection of the ctDNA in these samples and require less expensive DNA sequencing. In summary, the project should help make successful analysis of cancer patient's blood samples cheaper and more reliable, so reducing the number of biopsy operations that are required and enabling more evidence-based cancer therapy.