Identifying determinants of ADAR-dependency in triple-negative breast cancer

确定三阴性乳腺癌 ADAR 依赖性的决定因素

• 批准号: 10532305
• 负责人: Kyle Cottrell
• 金额: $ 0.81万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10532305
• 关键词: Address; Adenosine; Affect; African; African American; BRCA mutations; Biological; Biological Assay; Black race; Breast Cancer Cell; Breast Cancer Model; Breast Cancer cell line; Cell Line; Cell Proliferation; Cells; Classification; Clinical Data; DNA; Data; Data Correlations; Deaminase; Deamination; Dependence; Development; Disease; Disparity; Double-Stranded RNA; Enzymes; Exhibits; Gene Expression; Genes; Genetic Transcription; Grant; Growth; Inosine; Interferons; Lead; Left; Mammary Neoplasms; Modeling; Monitor; Mutation; Nucleotides; Oranges; Pathogenicity; Pathway interactions; Patient-derived xenograft models of breast cancer; Patients; Phenotype; Poly(ADP-ribose) Polymerase Inhibitor; Process; Proliferating; Proteins; RNA; RNA Folding; Research Personnel; Signal Transduction; Structure; Survival Rate; Testing; Therapeutic; Therapeutic Intervention; Training; Woman; Work; bench to bedside; career development; chemotherapy; disparity reduction; dsRNA adenosine deaminase; experience; genomic data; high throughput screening; immunogenic; implantation; inhibitor; knock-down; malignant breast neoplasm; mortality; mouse model; new therapeutic target; outcome disparities; patient derived xenograft model; patient population; performance tests; racial disparity; research and development; side effect; small molecule; small molecule inhibitor; small molecule libraries; targeted treatment; therapeutic target; treatment strategy; triple-negative invasive breast carcinoma; tumor; tumor growth; tumorigenesis

PROJECT SUMMARY Triple-negative breast cancer (TNBC), the deadliest form of breast cancer, affects Black/African American women at twice the rate of white women. Additionally, the survival rate for TNBC is lower in Black/African women. To address the disparities associated with TNBC we need broadly applicable targeted therapies. Without targeted therapies, clinicians are left with chemotherapy, which has many negative side- effects and in many cases is ultimately ineffective in the treatment of TNBC. We have observed that a subset of TNBC cell lines are dependent on the expression of Adenosine Deaminase Acting on RNA (ADAR). ADAR is an enzyme that converts adenosine nucleotides in RNA to inosine in a process known as A-to-I editing. Loss of ADAR inhibits cellular proliferation and tumor formation for a subset of TNBC cell lines. Because ADAR is required for the growth of some TNBC cell lines it serves as a valuable therapeutic target for the treatment of TNBC. It has been observed that ADAR-dependent cell lines have elevated interferon signaling, potentially making it possible to classify ADAR-dependent tumors. Interferon signaling is higher in Black/African American breast tumors than tumors in white patients, which may make therapies targeting ADAR more effective for Black/African American patients. There are aspects of ADAR-dependence that we do not understand. For instance, we do not understand why some cells are dependent on ADAR expression while others are not. Here we will explore the mechanism of ADAR-dependency and develop a strategy for treatment of TNBC based on ADAR inhibition. In AIM 1 we will identify the factors that are required for ADAR-dependence, including identification of the immunogenic RNAs that contribute to the phenotype caused by ADAR depletion. In AIM 2 we will develop and assess a classification model to predict which tumors will be sensitive to ADAR inhibition. The accuracy of the classification model will be evaluated by knockdown of ADAR in patient derived xenograft models of TNBC. Importantly this AIM will provide the PI with training in mouse models of breast cancer, including tumor implantation and monitoring. Finally, in AIM 3, we will develop a high-throughput A-to-I editing assay and use it to identify a small molecule inhibitor of ADAR. In addition to the potential identification of a small molecule inhibitor of ADAR, this AIM will provide the PI with experience developing a high-throughput screen and the use of DNA-encoded chemical libraries. This work will advance our understanding of ADAR- dependency such that we can accurately classify ADAR-dependent TNBC, thus opening the door to treating this deadly form of breast cancer with the small molecules identified in AIM 3. Developing an effective targeted therapy for TNBC is essential to reducing the disparate effects of this disease on Black/African American women. Finally, the research and career development training included in this grant will facilitate the PIs transition into an independent investigator.

项目摘要 三阴性乳腺癌（TNBC）是最致命的乳腺癌形式，影响了黑人/非洲 美国妇女的妇女率是白人妇女的两倍。此外，TNBC的生存率较低 黑人/非洲妇女。为了解决与TNBC相关的差异，我们需要广泛适用的目标 疗法。没有靶向疗法，临床医生将接受化学疗法，其中有许多负面的侧面 影响和在许多情况下最终在治疗TNBC方面无效。我们已经观察到一个子集 TNBC细胞系的依赖于作用于RNA（ADAR）的腺苷脱氨酶的表达。阿达 是一种将RNA中腺苷核苷酸转化为肌苷的酶，在称为A到I编辑的过程中。损失 ADAR抑制TNBC细胞系的细胞增殖和肿瘤形成。因为Adar是 某些TNBC细胞系生长所必需的，它是治疗的有价值的治疗靶标 TNBC。已经观察到ADAR依赖性细胞系具有升高的干扰素信号传导，可能 可以对ADAR依赖性肿瘤进行分类。干扰素信号在黑人/非裔美国人中更高 乳腺肿瘤比白人患者的肿瘤，这可能会使针对ADAR的疗法更有效 黑人/非裔美国人患者。我们不了解ADAR依赖性的某些方面。为了 实例，我们不明白为什么有些单元格依赖于ADAR表达而而其他细胞则不依赖。这里 我们将探讨ADAR依赖性的机制，并制定基于TNBC治疗的策略 ADAR抑制。在AIM 1中，我们将确定ADAR依赖性所需的因素，包括 鉴定有助于ADAR耗竭引起的表型的免疫原性RNA。在目标2中 我们将开发和评估分类模型，以预测哪些肿瘤将对ADAR抑制敏感。 分类模型的准确性将通过在患者衍生的异种移植物中敲除ADAR来评估 TNBC的模型。重要的是，此目标将为PI提供乳腺癌模型的培训， 包括肿瘤植入和监测。最后，在AIM 3中，我们将开发一个高通量A到I编辑 测定并使用它来识别ADAR的小分子抑制剂。除了潜在的识别 ADAR的小分子抑制剂，此目标将为PI提供高通量的经验 屏幕和使用DNA编码的化学文库的使用。这项工作将促进我们对Adar的理解 依赖性使我们可以准确地对ADAR依赖性TNBC进行分类，从而打开了治疗的大门 这种致命的乳腺癌形式，在AIM 3中鉴定出小分子。开发有效的靶向目标 TNBC的治疗对于降低这种疾病对黑人/非裔美国人的不同影响至关重要 女性。最后，本赠款中包括的研究和职业发展培训将有助于PIS 过渡到独立研究者。