Development and clinical validation of a multi-type HPV E6/E7 oncoprotein test for cervical cancer screening and triage in low- and middle-income countries

 描述（由申请人提供）：宫颈癌仍然是大多数低收入和中等收入国家（LMIC）癌症发病率和死亡率的主要原因。尽管在发达国家，基于细胞学的筛查计划已将宫颈癌死亡率降低至80%，但这些计划在发展中国家经常失败，主要是由于需要多次访问以确定需要治疗的风险妇女。宫颈细胞学检查的敏感性较低（约50%），因此需要通过终身筛查频繁重复检查;此外，阴道镜活检的补充诊断要求妇女在短时间内多次就诊，以接受适当的治疗。最近，FDA开发并批准了检测致癌HPV的高灵敏度和可重复性实验室技术，作为宫颈癌的主要筛查试验。随机临床试验表明，HPV检测比细胞学检测宫颈癌前体更敏感（约90%对50%），并且HPV检测在早期阶段检测到更多疾病，并导致宫颈癌死亡率大幅降低。这些技术正在改变墨西哥、阿根廷、哥伦比亚和萨尔瓦多等中低收入国家的筛查做法。尽管有其优点，HPV检测具有较低的阳性预测值;因此，它们需要额外的诊断程序（分诊），以避免阴道镜诊所的负担过重或过度治疗，如果没有进行确诊（见和治疗计划）。目前，对阳性妇女的分类是用细胞学进行的，由于敏感性低，细胞学再次产生了高召回率，这对低资源环境下的筛查计划提出了挑战。目前，没有一个正在调查的分类测试是适合筛选程序在LMIC由于诱导高召回率或高物流或基础设施的要求，其实施。以友好/手动格式开发高度准确的测试，以个人为基础产生结果，将有助于改善低资源环境中筛查计划的结果。我们将利用拉丁美洲国家已经建立的宫颈癌研究网络的平台：（目标1）开发和改进基于E6/E7癌蛋白活性的HPV检测，用于与宫颈癌相关的8种最常见的HPV类型;（目的2）评估8-HPV E6/E7型的潜在临床性能，通过对800份样本的整个疾病组织学诊断谱进行挑战，（目的3）评估ESTAMPA研究中4,500名女性HPV阳性女性（分诊）中癌症前体检测测试的灵敏度和特异性;（目的4）使用额外1，000名HPV阴性女性的样本，评估该测试作为子宫颈癌筛查的独立测试的潜力;（目的5）比较（阳性/阴性）的8-HPV E6/E7型为基础的OncoProtein Cervical Test与HPV持续存在和发展的宫颈病变后18个月的妇女随访;（目标6）通过在拉加经委会三个不同实验室处理4，500个样本，确定不同环境下检测性能的差异;（目标7）确定扩大基于8-HPV E6/E7型癌蛋白宫颈检测的宫颈癌筛查计划的操作要求，在5个不同的LAC中真实的500份样本;（目的8）描述了包含HPV 16和18癌蛋白的origina OncoE 6 TM宫颈检测试剂盒与基于新的8-HPV E6/E7型癌蛋白宫颈检测试剂盒之间的灵敏度和特异性差异。

 DESCRIPTION (provided by applicant): Cervical cancer remains a leading cause of cancer incidence and mortality in most low and middle income countries (LMIC). Despite cytology based screening programs have reduce cervical cancer mortality up to 80% in developed nations, these programs have frequently failed success in developing countries mainly due to the requirement of multiple visits to identify women at risk who require treatment. The low sensitivity of cervical cytology (around 50%) induces the frequent repetition of the test through lifetime screening; in addition, the complementary diagnosis with colposcopy-biopsy requires women to attend multiple visits in the short time to undergo proper treatment. Recently, highly sensitive and reproducible laboratory techniques to detect carcinogenic HPV have been developed and approved by FDA as primary cervical cancer screening tests. Randomized clinical trials have demonstrated that HPV testing is more sensitive than cytology to detect cervical cancer precursors (around 90% vs 50%) and it has also been shown that HPV testing detects more disease at an earlier stage and induces a greater reduction in cervical cancer mortality. These technologies are changing screening practices in low and middle income countries such as Mexico, Argentina, Colombia, and El Salvador. Despite its advantages, HPV tests have a low positive predictive value; thus, they require additional diagnostic procedures (triage) to avoid overburden of colposcopy clinics or overtreatment if no confirmatory diagnosis is performed (see-and-treat programs). Currently, the triage for positive women is done with cytology, which again, due to the low sensitivity produces a high recall rate challenging screening programs in low resource settings. At the moment none of the triage tests under investigation is suitable for screening programs in LMIC due to the induction of high recall rates or high logistic or infrastructure requirements for its implementation. The development of a highly accurate test in a friendly/manual format producing results on individual basis will help improve outcomes of screening programs in low resource settings. We will use the platform of a cervical cancer research network already in place in Latin American countries (LAC) to: (Aim 1) developed and improved HPV test based on E6/E7 oncoprotein activity for the 8 most common HPV types associated with cervical cancer; (Aim 2) assess the potential clinical performance of the 8-HPV type E6/E7-based OncoProtein Cervical Test by challenging it against the whole spectrum of disease's histological diagnosis in 800 samples; (Aim 3) asses the sensitivity and specificity of the test for the detection of cancer precursors among HPV positive women (triage) in 4,500 women from the ESTAMPA study; (Aim 4) assess the potential of the test as standalone test for cervical cancer screening by using a sample of additional 1,000 HPV negative women; (Aim 5) to compare the results (positive/negative) of the 8-HPV type E6/E7-based OncoProtein Cervical Test with HPV persistence and development of cervical lesions after 18 months of women follow-up; (Aim 6) identify differences in the performance of test in different settings by processing 4,500 samples in three different labs in LAC; (Aim 7) identify operational requirements for scaling up cervical cancer screening programs based on the 8-HPV type E6/E7-based OncoProtein Cervical Test by performing 1,500 samples in real time in five different LAC; (Aim 8) describe differences in sensitivity and specificity between the origina OncoE6TM Cervical Test that included oncoproteins for HPV 16 and 18 and the new 8-HPV type E6/E7-based OncoProtein Cervical Test.