Impact of preanalytic procurement and processing variables on the detection of HCC DNA in urine

分析前采购和加工变量对尿液中肝细胞癌DNA检测的影响 肝细胞癌是世界上第二大癌症相关死亡原因，也是 美国增长最快的癌症；85%的患者在5年内死亡，主要是由于发现较晚，有限 治疗选择多，复发率高。目前，正在监测肝细胞癌复发和治疗反应。 通过血清甲胎蛋白和系列影像检查。不幸的是，血清甲胎蛋白只能用于约50%的肝细胞癌患者 低敏感度。虽然MRI/CT成像是诊断的黄金标准，但它很昂贵，而且更难获得。 它在检测小肿瘤(&lt；2厘米)方面的效用也是有限的，而且在存在 发育不良的结节。用目前可用的方法，早期发现复发的肝细胞癌是无效的。遗传 液体活组织检查已经在临床上被常规用于几乎所有类型的癌症，作为国家 综合癌症网络(NCCN)指南。然而，它在肝癌中的使用是有限的，尽管它的 承诺在文献中得到了很好的记录，主要是关于血液，也有一些是关于尿液的。我们的假设 是通过建立以证据为基础的尿液收集、储存和跨肾生物检疫实践 DNA(TrDNA)分离、尿中肝细胞癌DNA分析和特制灵敏的分析方法可为肝细胞癌遗传学提供 与目前的护理标准相比，更频繁地进行肿瘤监测和治疗反应评估 用来判断预后。作为发展尿道癌液体活检的先驱，我们已经建立了标准 结肠癌早期检测研究网络尿液采集操作规程 验证研究(方案ID：320)和其他非尿路癌症。我们已经确定了分析前 肝细胞癌尿DNA制备过程中影响尿肝癌DNA检测性能的变量 肝细胞癌液体活组织检查。在这项应用中，我们建议使用肝脏特异性trDNA标志物，乙肝 从慢性乙肝患者的尿液中发现的病毒(乙肝病毒)DNA，以验证和缓解 这些分析前的变量。目标1和目标2将识别、验证和缓解分析前因素，相关 通过尿液的采集、储存和加工分离trDNA，并表征trDNA的大小， 影响临床化验结果和重复性。因此，一种循证的尿液生物检验剂 可以建立实践来帮助支持尿液中肝细胞癌DNA检测的开发、对肝细胞癌基因的验证 液体活组织检查和治疗反应评估。在目标3中，我们将演示可重复性 以及临床验证尿液中肝细胞癌DNA在评估肝细胞癌治疗反应中的作用。发展中 用于样本收集、处理和长期存储的可靠的循证尿液生物检验法 将导致临床试验中用于评估肝细胞癌治疗的尿液测试的有效性。

Impact of pre-analytic procurement and processing variables on the detection of HCC DNA in urine Hepatocellular carcinoma (HCC) is the world’s 2nd leading cause of cancer-related death and one of the fastest-growing cancers in the US; 85% of patients die within 5 years, mainly due to late detection, limited treatment options, and high recurrence. Currently, HCC recurrence and treatment responses are monitored by serum AFP and serial imaging. Unfortunately, serum AFP can only be used for ~50% of HCC cases due to low sensitivity. While MRI/CT imaging is the gold standard for diagnosis, it is expensive and less accessible. It also has limited utility in the detection of small tumors (< 2 cm) and is challenging in the presence of dysplastic nodules. Early detection of recurrent HCC is ineffective with the current available methods. Genetic liquid biopsies have been routinely used in clinics for almost all cancer types as part of the National Comprehensive Cancer Network (NCCN) guideline. However, its usage in HCC is limited even though its promises have been well documented in the literature, mostly for blood and some for urine. Our hypothesis is that by establishing an evidence-based biospecimen practice for urine collection, storage, and transrenal DNA (trDNA) isolation, urine HCC DNA analysis with tailored sensitive assays can provide HCC genetics with more frequent tumor surveillance and treatment response assessment compared to current standard of care for prognosis. As a pioneer in developing urine cancer liquid biopsies, we have established the standard operating procedure (SOP) for urine collection for the Early Detection Research Network colon cancer validation study (protocol ID: 320) and other non-urinary tract cancers. We have identified pre-analytic variables that influence urine HCC DNA assay performance in the process of developing an HCC urine DNA test for HCC liquid biopsies. In this application, we propose to use a liver-specific trDNA marker, hepatitis B virus (HBV) DNA, which is found in urine, from patients with chronic HBV infection to validate and mitigate these pre-analytic variables. Aims 1 and 2, will identify, validate, and mitigate pre-analytic factors, associated with urine collection, storage and processing for trDNA isolation, and characterize of the size of trDNA that influences clinical assay results and reproducibility. As a result, an evidence-based urine biospecimen practice can be established to help support urine HCC DNA assay development, validation for HCC genetic liquid biopsy and assessment of treatment response. In Aim 3, we will then demonstrate the reproducibility and clinical validation of the utility of urine HCC DNA in assessing responses to HCC treatment. Developing a robust evidence-based urine biospecimen practice for sample collection, processing, and long-term storage will lead to the validation of urine tests for assessing HCC treatment in clinical trials.