Identifying biomarkers of treatment response in gestational trophoblastic disease through quantitative proteomics analysis of patient samples

妊娠滋养细胞疾病(GTD)是一组因卵子受精异常而引起的妊娠相关疾病。GTD在英国每年影响约1800名妇女，是一系列疾病，包括恶性葡萄胎(HM)到恶性绒毛膜癌。尽管GTD的临床治疗已经取得了很大进展，但仍有几个问题有待解决。特别是，由于缺乏足够的预测生物标记物，对最合适的治疗策略的知情选择受到了损害。确实是这样。虽然基于甲氨蝶呤(MTX)的单一疗法可以治愈绝大多数病例，但我们目前的分层评分系统(FIGO评分)对很大一部分患者进行了错误的分类，导致了治疗性联合化疗的不必要延误。此外，对联合药物治疗耐药的患者可能需要进一步的不同联合化疗，包括大剂量治疗和/或免疫检查点治疗，以实现持续缓解，但缺乏在这些治疗方法之间选择患者的生物标志物。因此，患者可能会经历多轮不同的治疗。如果我们有一个适当的分层生物标记物，这些以及相关的毒性可能会被避免。最后，尽管大多数患者最终被治愈，但一些人继续失败现有的所有治疗，并将受益于新疗法的新靶点的确定。在这里，我们打算使用临床注释的患者组织样本的蛋白质组学分析来识别更好的生物标记物，将GTD患者分层到现有的治疗方法中，并发现靶点(已知的或新的)以实现新的治疗策略。我们之前对单个MTX敏感/耐药绒毛癌细胞系的蛋白质组学分析显示，DNA修复和细胞周期检查点是解决MTX耐药性的潜在新治疗靶点。然而，在GTD组织样本中对这些发现的后续验证表明，这些发现只与一小部分患者相关。因此，需要一种不那么专注的方法来发现与单一化疗药物耐药与多药耐药相关的更多变化。具体地说，该项目的目标是：1.使用基于定量质谱学(MS)的GTD组织样本蛋白质组学分析来识别与不同的治疗耐药性特征相关的分子通路2.使用GTD细胞系在体外和体内确认所识别的通路在相关治疗方案的耐药性中的作用3.在大量组织样本中验证上述发现的临床相关性，以统计地评估这些生物标记物和治疗反应之间的关联强度

Gestational trophoblastic disease (GTD) is a group of pregnancy related disorders which arise after abnormal fertilization of the egg. GTD affects about 1800 women/yr in the UK and is a spectrum of disorders including the pre-malignant hydatidiform moles (HM) through to the malignant choriocarcinoma. Although much progress has been done in the clinical management of GTD, several problems remain to be addressed. In particular, informed choice of the most appropriate therapeutic strategy is impaired by the lack of adequate predictive biomarkers. Indeed. While methotrexate (MTX)-based monotherapy cures the vast majority of cases, our current stratification scoring system (FIGO score) misclassifies a large proportion of patients, leading to unnecessary delays in administration of curative combination chemotherapy. In addition, patients resistant to combination agent treatment may require further different combination chemotherapy including high dose treatment and/or immune-checkpoint therapy to achieve a sustained remission but biomarkers to select patients between these therapies are lacking. Thus, patients may go through multiple rounds of different treatments. These could potentially have been avoided along with the associated toxicities if we had an appropriate stratification biomarker. Finally, although most patient eventually get cured, some continue to fail all existing treatments and would benefit from the identification of novel targets for new therapies. Here, we intend to use proteomics profiling of clinically annotated patient tissue samples to identify better biomarkers for stratification of GTD patients into existing treatments and discover targets (known or novel) to enable new therapeutic strategies. Our previous proteomics profiling of a single MTX-sensitive/resistant choriocarcinoma cell line pair revealed DNA repair and cell cycle checkpoints as potential new therapeutic targets to tackle MTX resistance. However, follow-on validation of these findings in GTD tissue samples showed these to be only relevant to a subset of patients. Therefore, a less focused approach is needed to discover more changes associated with resistance to single versus multi-chemotherapeutic agents. Specifically, the aims of this project are: 1. Identification of molecular pathways associated with distinct therapy resistance profiles using quantitative mass-spectrometry (MS)-based proteomics analysis of GTD tissue samples 2. Confirmation of the role of the identified pathways in the resistance to the relevant therapeutic regimen in vitro and in vivo using GTD cell lines 3. Validation of the clinical relevance of the above findings in a large number of tissue samples to statistically assess the strength of association between these biomarkers and therapeutic response