Pancreatic Cyst Fluid miRNOME for Biomarkers of Pancreatic Cancer

 描述（由申请人提供）：胰腺导管腺癌（PDAC）是一种高致死性癌症，平均5年生存率<6%，因为大多数患者在诊断时存在局部或远端转移，而治愈性治疗选择极其有限。在超过2%的一般患者人群中通过腹部成像检测到的无症状胰腺囊肿为开发PDAC的早期检测生物标志物提供了独特的机会，因为分泌粘蛋白的胰腺囊肿，例如更常见的导管内乳头状粘液性肿瘤（IPMN）和不太常见的粘液性囊性肿瘤（MCN）是PDAC的真正前驱病变。此外，来自粘液性囊性病变的浸润性癌症的生物标志物预计将解决可靠区分惰性与侵袭性囊性病变的关键未满足的临床需求，以帮助惰性疾病患者避免手术相关的发病率和死亡率的风险。在最近的一项有限样本量队列研究中，我们通过NextGen测序（NGS）鉴定了来自高级别IPMN浸润性胰腺癌患者与低级别IPMN患者的差异丰富的囊液miR（cf-miR），证明了开发cf-miR标记作为早期检测生物标志物的可行性，用于将倾向于从低风险惰性囊肿进展为浸润性PDAC的高风险IPMN分层（Wang等人Cancer Letters 2015; 356：404-409）。在该项目中，我们提出对来自具有良性或低度异型增生的IPMN患者和具有高度异型增生-浸润性癌症的IPMN患者的NGS衍生的囊液miRNome（cf-miR）进行详细分析，以开发可以区分侵袭性IPMN与良性和惰性病变的cf-miR特征。此外，我们建议研究与PDAC相关的IPMN患者的cf-miR和血浆miR特征之间的重叠程度。本研究将利用我的合作者Kimberly柯克伍德博士和Pamela巴黎博士在UCSF Helen Diller Family Comprehensive Cancer Center进行的诊断性囊性取样（EUS）进行IPMN诊断和IPMN相关组织学组织和生物标本库（请参见随附信函）。我们提出以下具体目标： 目的1：利用NGS和qRT-PCR开发用于IPMN风险分层的cf-miRNA生物标志物签名。 目标二：研究在切除的组织样品中和在表达非致瘤性人胰腺导管上皮（HPDE）细胞的突变K-RasG 12 V的体外细胞系模型中差异丰度的cf-miR的功能意义。 目的3：评价胰腺囊肿患者血浆中cf-miRNA生物标志物特征。

 DESCRIPTION (provided by applicant): Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal cancer with an average 5 year survival rate of <6%, since most patients present with local or distal metastasis at the time of diagnosis when curative treatment options are extremely limited. Asymptomatic pancreatic cysts detected in more than 2% of the general patient population with abdominal imaging presents a unique opportunity for developing early detection biomarkers of PDAC since mucin-secreting pancreatic cysts, such as the more frequently detected intraductal papillary mucinous neoplasms (IPMNs) and the less frequent mucinous cystic neoplasms (MCNs) are bona fide precursor lesions of PDAC. Furthermore, biomarkers of invasive cancer from mucinous cystic lesions is expected to address the critical unmet clinical need of reliably distinguishing indolent from aggressive cystic lesions to help patients with indolent disease avoid the risk of surgery associated morbidity and mortality. In a recent limited sample size cohort study, we identified by NextGen sequencing (NGS) differentially abundant cyst fluid miRs (cf-miR) from patients with high grade IPMN-invasive pancreatic cancer compared with those with low grade IPMN demonstrating the feasibility of developing cf-miR signatures as early detection biomarkers for stratifying high risk IPMN predisposed to progress to invasive PDAC from low risk indolent cysts (Wang et al. Cancer Letters 2015; 356: 404-409). In this project, we are proposing to undertake detailed analyses of NGS derived cyst fluid miRNome (cf-miR) from IPMN patients with benign or low grade dysplasia and those with high grade dysplasia- invasive cancer to develop cf-miR signature that can differentiate aggressive IPMN from benign and indolent lesions. Additionally, we propose to investigate extent of overlap between the cf-miR and the plasma miR signatures from IPMN patients associated with PDAC. The study will take advantage of diagnostic cystic sampling with endoscopic ultrasound (EUS) for IPMN diagnosis and IPMN related histological tissue and bio-specimen banking at the Helen Diller Family Comprehensive Cancer Center at UCSF by my collaborators, Dr. Kimberly Kirkwood and Dr. Pamela Paris respectively (Please see the letters attached). We propose the following specific aims: Aim 1: Develop cf-miRNA biomarker signature for IPMN risk stratification utilizing NGS and qRT-PCR. Aim 2: Investigate functional significance of the differentially abundant cf-miRs in resected tissue samples and in an in vitro cell line model of mutant K-RasG12V expressing non-tumorigenic human pancreatic ductal epithelial (HPDE) cells. Aim 3: Evaluate cf-miRNA biomarker signatures in blood plasma from patients with pancreatic cysts.