Identifying Novel Radiation Sensitizers in Cervical Cancer

鉴定宫颈癌的新型放射增敏剂

• 批准号: 10577052
• 负责人: Lauren Elizabeth Colbert
• 金额: $ 22.72万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-07 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10577052
• 关键词: Address; Aftercare; Biological Models; Biopsy; Blood Banks; CRISPR library; Cancer Patient; Cancer cell line; Candidate Disease Gene; Cause of Death; Cell Line; Cervical; Chemotherapy and/or radiation; Clinic; Clinical; Clonal Expansion; Custom; DNA Library; DNA Sequence Alteration; Data; Databases; Dimensions; Drug Screening; Evolution; Fistula; Freezing; Fresh Tissue; Future; Gene Pool; Genes; Hemorrhage; Immune; In Vitro; Institution; Laboratories; Libraries; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Methods; Modeling; Mutate; Mutation; Organoids; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacotherapy; Pilot Projects; Population; Precision therapeutics; Prognostic Marker; Publishing; Qualifying; Radiation-Sensitizing Agents; Radiosensitization; Recording of previous events; Relapse; Resistance; Risk; Sampling; Swab; Techniques; Testing; Tissue Banks; Tissues; Translating; Treatment outcome; Woman; biobank; candidate validation; cell bank; chemoradiation; clinical translation; cohort; computational pipelines; driver mutation; exome sequencing; experience; genetic testing; high risk; human tissue; improved; in vitro testing; large-scale database; microbiome; new therapeutic target; novel; patient population; predictive marker; radiation resistance; radiation response; research clinical testing; response; risk variant; screening; single-cell RNA sequencing; standard of care; targeted agent; targeted treatment; therapeutically effective; tumor; tumorigenesis

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.

项目摘要 宫颈癌仍然是全球女性死亡的一个重要原因，40%的女性患有宫颈癌， 会复发并在放化疗后死亡迫切需要确定预后和 放化疗反应的预测性生物标志物。我们开发并优化了一种新型的非侵入性 基于拭子的活检方法在放化疗之前、期间和之后收集组织，而没有患者不适 或连续活检的风险，如出血或瘘管。尽管肿瘤纯度很低，我们还开发了一种定制的 计算流水线，以优化突变调用并识别克隆扩增的突变， 放化疗根据这些发现，深入了解CRT期间和之后的基因组改变 将有助于靶向治疗的快速临床转化。为此，我们已利用我们的管道进行了一项试验研究 重建CRT期间突变的演变，并确定增殖优势 驱动突变和途径。该提案旨在验证多个候选驱动程序突变列表 尺寸和开发一个患者源性类器官平台，以测试潜在的靶向治疗。一是 验证初步确定的克隆扩增的基因和途径，在更大的人口已经 已经收集但尚未测序的样本第二，我们将对这些基因进行CRISPR/Cas9文库筛选， 在可用的宫颈癌细胞系中初步鉴定了驱动因子。与此同时，我们将开发一个颈椎 癌症类器官生物库在CRT前后进行单细胞RNA测序，并验证这些 初步鉴定基因。该平台将在未来用于测试目标代理。当我们完成 为了实现R21的目标，我们将验证理想目标，开发测试平台，并为 在未来的R 01中测试具有临床影响力的疗法。