Identification and validation of circulating sncRNAs causally associated with gallbladder cancer and development of a multifactorial risk prediction score

每年约有57,000人死于胆囊癌(GBC)。早期症状模糊，确诊五年后只有5%的GBC患者还活着。胆囊癌的手术切除提供了治愈局限于粘膜的胆囊癌的可能性。不幸的是，目前还没有可靠的测试来预测或检测早期胆汁淤积症。这项研究的目的是：1)揭示GBC发生的原因机制；2)在临床表现之前，开发和验证GBC预测的风险评分，最终彻底改变这种被忽视的疾病的预防。该项目头三年的具体目标是：1)根据“胆结石和炎症-异型增生-GBC”序列所代表的多阶段致癌模型中胆囊壁组织的表达变化，预先选择候选的SncRNAs；2)识别和验证与GBC风险相关的循环中的SncRNAs；3)测试因果关系，并通过孟德尔随机化方法估计相关SncRNAs的因果效应；以及4)建立风险评分，将已建立的GBC风险因素和新发现的与GBC风险相关的SNcRNAs结合起来。全面的临床和环境暴露信息，以及来自90个福尔马林固定石蜡包埋(FFPE)组织样本(30个来自胆结石，30个来自异型增生，30个来自GBC智利患者)的SNcRNA序列数据。这一数据集将补充临床和环境数据，以及·另外180份来自秘鲁和阿根廷患者的FFPE样本(60名胆结石患者，60名不典型增生患者和60名GBC患者)，以及·375名胆结石和375名GBC患者的独立血清/DNA样本(智利250名，秘鲁250名，阿根廷250名)。GBC相对罕见--它每年在德国造成约1500人死亡(krebsdaten.de)。这种肿瘤侵袭性很强，但有足够的机会预防胆囊癌：肿瘤的发展需要10-20年，对胆囊癌高危人群可以预防性切除。对GBC形成的因果机制的新见解将提高目前对已建立的风险因素和GBC之间生物学联系的理解，最终将相关发现转化为一般公共卫生政策，特别是GBC预防。根据制定的风险评分对胆囊癌风险的个人估计将指导监测和个人决定预防性胆囊癌切除术对高危人群(例如胆囊癌患者的一级亲属和严重肥胖妇女)可能带来的好处。这将大大改善工业化国家和欠发达国家的GBC预防工作。从GBC发病率较高的区域获得独特的数据集和样本收集，以及使用创新的实验室技术和先进的统计方法，有力地支持了本提议。

Around 57,000 persons die every year due to gallbladder cancer (GBC). Early symptoms are vague and only 5% of GBC patients are alive five years after diagnosis. Surgical removal of the gallbladder(cholecystectomy) offers the potential for cure of GBC confined to the mucosa. Unfortunately, a reliable test to predict or detect early GBC is not available yet. This research proposal aims at 1) unravelling the causal mechanisms underlying GBC development, and 2) developing and validating a risk score for GBC prediction before clinical manifestation, eventually revolutionizing the prevention of this neglected disease. Specific objectives for the first three years of the project are: 1) to pre-select candidate sncRNAs based on expression changes in gallbladder tissue along the multistage model of carcinogenesis represented by the sequence “Gallstones and Inflammation – Dysplasia – GBC”, 2) to identify and validate circulating sncRNAs associated to GBC risk, 3) to test the causality, and to estimate the causal effects of associated sncRNAs by Mendelian randomization, and 4) to build a risk score which combines established GBC risk factors and newly identified sncRNAsassociated with GBC risk. Comprehensive clinical and environmental exposure information, and sncRNA sequence data from 90 formalin-fixed paraffin-embedded (FFPE) tissue samples (30 from gallstone, 30 from dysplasia and 30 from GBC Chilean patients) are available. This dataset will be complemented with clinical and environmental data, and with • 180 additional FFPE samples from Peruvian and Argentinian patients (60 with gallstones, 60 withdysplasia and 60 affected by GBC), and • independent serum/DNA samples from 375 gallstone and 375 GBC patients (250 from Chile, 250 from Peru and 250 from Argentina). GBC is relatively rare – it causes around 1500 deaths in Germany every year (krebsdaten.de). The tumor is very aggressive but there is ample opportunity for GBC prevention: 10-20 years are needed for tumour development and prophylactic cholecystectomy can be offered to individuals at high GBC risk. New insights into the causal mechanisms that underlie GBC development will improve current understanding of the biological links between established risk factors and GBC, eventually translating into relevant findings to public health policies in general, and to GBC prevention in particular. Individual estimates of GBC risk based on the developed risk score would guide surveillance and personal decisions on the possible benefit of prophylactic cholecystectomy for persons at high risk (e.g., first-degree relatives of GBC patients and severely obese women). This would significantly improve GBC prevention in both industrialized and less developed countries. The access to unique datasets and sample collections from regions with a high GBC incidence, and the use of innovative laboratory techniques and advanced statistical methods strongly endorse the present proposal.