Identifying Novel Radiation Sensitizers in Cervical Cancer

项目摘要 宫颈癌仍然是全球妇女死亡的重要原因，40%患有宫颈癌的妇女 即使接受放化疗也会复发和死亡。迫切需要确定预后和 化疗放射反应的预测生物标志物。我们已经开发和优化了一种新型的、非侵入性的 以拭子为基础的活检方法，在化疗前、中和之后收集组织，而不会使患者感到不适 或连续活检的风险，如出血或瘘管。尽管肿瘤纯度较低，但我们也发展了一种习俗 优化突变调用并识别克隆扩展突变的计算流水线 放化疗。加深对CRT术中和术后基因组改变的理解 将有助于快速跟踪靶向治疗的临床翻译。为此，我们利用我们的渠道进行了一项初步研究 在70名患者中重建CRT期间突变的演变并确定增殖优势 驱动基因突变和路径。该方案旨在对多个候选司机突变列表进行验证 测量并开发患者衍生的有机类药物平台，以测试潜在的靶向治疗。首先，我们将 在一个更大的种群中验证初步鉴定的克隆扩展的基因和途径 收集的但尚未测序的样本。其次，我们将执行以下内容的CRISPR/Cas9库筛选 初步确定了可用的宫颈癌细胞系中的驱动因素。同时，我们将开发一种宫颈 癌症有机体生物库在CRT前后进行单细胞RNA测序并验证 初步确定了基因。该平台将在未来用于测试目标代理。当我们完成时 R21的目标，我们将验证理想的目标，开发测试平台，并为 在未来的R01中测试临床有效的治疗方法。

Project Summary Cervical cancer remains a significant global cause of death in women, and 40% of women with cervical cancer will relapse and die despite chemoradiation treatment. There is an urgent need to identify prognostic and predictive biomarkers for chemoradiation response. We have developed and optimized a novel, non-invasive swab-based biopsy approach to collect tissue before, during, and after chemoradiation without patient discomfort or risks of serial biopsies such as bleeding or fistula. Despite low tumor purity, we have also developed a custom computational pipeline to optimize mutation calling and identify clonally expanded mutations during chemoradiation. Developing a deep understanding of genomic alterations during and after CRT to these findings will help fast-track clinical translation of targeted therapies. To this end, we have used our pipeline in a pilot study of 70 patients to reconstruct the evolution of mutations during CRT and identify proliferatively advantageous driver mutations and pathways. This proposal aims to validate the candidate driver mutation lists in multiple dimensions and develop a patient-derived organoid platform to test potential targeted therapies. First, we will validate the preliminary identified clonally expanded genes and pathways in a larger population of already collected but yet-to-be-sequenced samples. Second, we will perform a CRISPR/Cas9 library screen of these preliminarily identified drivers in available cervical cancer cell lines. Simultaneously, we will develop a cervical cancer organoid biobank to perform single-cell RNA sequencing before and after CRT and validate these preliminarily identified genes. This platform will be used in the future to test targeted agents. When we complete the aims of this R21, we will have validated ideal targets, developed testing platforms, and set the stage for testing clinically impactful therapies in a future R01.