The Role of PAQR8 in Breast Cancer Metastasis and Therapy Resistance

PAQR8 在乳腺癌转移和治疗耐药中的作用

基本信息

批准号：
10251898
负责人：
Saisai Chen
金额：
$ 3.34万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10251898
关键词：
Address Animals Antiestrogen Therapy Apoptosis Bilateral Binding Biological Assay Breast Cancer Cell Breast Cancer Patient Breast Cancer cell line Breast cancer metastasis CRISPR/Cas technology Cancer Etiology Cause of Death Cells Cessation of life Code Complement Data Diagnosis Dose ERBB2 gene Early treatment Estrogen Antagonists Estrogen Receptors Exons Family Fatty acid glycerol esters Frequencies Genes Genetically Engineered Mouse Hormonal Hormone Receptor Hormones Human Image Immunocompromised Host Immunofluorescence Immunologic In Vitro Interleukin 2 Receptor Knock-out Label Laboratories Lesion Light Luciferases MAP Kinase Gene MAPK8 gene MYC gene Mediating Mediator of activation protein Membrane Metastatic to Metastatic/Recurrent Monitor Mus Mutation Neoplasm Metastasis Nuclear Oncogenes Pathway interactions Patients Play Primary Lesion Primary Neoplasm Progesterone Progesterone Receptors Prognosis Proto-Oncogene Proteins c-akt RNA Recurrence Recurrent disease Recurrent tumor Residual Tumors Residual state Resistance Role Signal Pathway Signal Transduction Site Stains Time Transplantation Woman breast cancer progression cancer recurrence cancer therapy cell type cellular transduction chemotherapy early screening exome sequencing experimental study hormone therapy in vivo insight malignant breast neoplasm mammary member mortality new therapeutic target novel overexpression p38 Mitogen Activated Protein Kinase receptor targeted treatment tumor tumor growth tumorigenesis vector control

项目摘要

Project Summary Despite recent advances in diagnosis and treatment, breast cancer remains the most common cause of cancer-related deaths among women globally. Most deaths from breast cancer are due to metastatic and recurrent disease that occurs years after primary tumor regression from treatment. Consequently, there is a pressing need to better understand the mechanisms that underly metastasis and therapy resistance. Recently, our laboratory performed whole-exome sequencing of both matched metastatic and primary tumors and unmatched metastatic tumors from breast cancer patients. We identified copy number (CN) gain of a 77.5kbp region encompassing the full exon sequence of PAQR8 that was enriched in the metastases compared to primary tumors. PAQR8 encodes a membrane progesterone receptor (mPR) that has never before been implicated in breast cancer metastasis or recurrence. Moreover, CN gain of this region was mutually exclusive of mutations in estrogen receptor (ESR1) and progesterone receptor (PGR) among patients who received anti- estrogen therapy. But intriguingly, it was found in equal frequency among patients who were treated with non- hormonal therapies as those with hormonal therapy. Plus, both CN gain and increased RNA levels of Paqr8 were identified in recurrent compared to primary tumors of several genetically engineered mouse (GEM) models of oncogene-driven breast cancers, none of which express hormone receptors. Given these findings, I hypothesize that PAQR8 gain promotes breast cancer metastasis and therapy resistance. Aim 1 addresses whether PAQR8 gain promotes metastasis. I will orthotopically transplant an equal mixture of PAQR8- overexpressing and control cells into immunocompromised mice and track the formation of metastatic lesions by in vivo luciferase imaging. Comparing the ratio of the two different cell types in metastatic and primary lesions, I expect to find an enrichment for PAQR8-overexpressing cells in the metastases compared to primary tumors. Next, I will use immunofluorescence to detect active mediators predicted to be downstream of PAQR8 signaling in both primary and metastatic lesions. Aim 2 addresses whether PAQR8 gain promotes therapy resistance. I will perform a recurrence free survival assay to determine whether mice injected with PAQR8- overexpressing cells recur earlier following treatment than those injected with control cells. To investigate the composition of residual and recurrent tumors, I will orthotopically inject a mixture of PAQR8-overexpressing and control cells as in aim 1. Primary tumors will be treated with targeted therapy or chemotherapy, and both residual lesions and recurrent tumors will be assayed for the ratio of PAQR8-overexpressing cells to controls. I expect PAQR8-overexpressing cells to be enriched in both the residual and recurrent tumors. Finally, an in vitro cell competition assay will determine whether PAQR8 overexpression mediates therapy resistance in a progesterone dose-dependent manner. Results from the proposed experiments will yield novel insight into the role of PAQR8 membrane progesterone receptor in breast cancer metastasis and therapy resistance.

项目摘要尽管诊断和治疗最近取得了进步，但乳腺癌仍然是全球女性与癌症有关的死亡。大多数乳腺癌死亡都是由于转移性和原发性肿瘤在治疗后几年发生的复发性疾病。因此，有一个迫切需要更好地了解基本转移和耐药性的机制。最近，我们的实验室对匹配的转移性和原发性肿瘤进行了全外观测序，来自乳腺癌患者的无与伦比的转移性肿瘤。我们确定了77.5kbp的副本号（CN）增益与与转移中富集的PAQR8的完整外显子序列相比，原发性肿瘤。 PAQR8编码从未有过的膜孕酮受体（MPR）与乳腺癌转移或复发有关。而且，该地区的CN增益是相互排斥的接受抗抗雌激素受体（ESR1）和孕激素受体（PGR）的突变雌激素疗法。但是有趣的是，在接受非 - 激素疗法作为荷尔蒙治疗的疗法。另外，CN增益和RNA升高PAQR8 与几种基因工程小鼠（GEM）的原发性肿瘤相比，在复发中鉴定出来致癌基因驱动的乳腺癌的模型，均未表达激素受体。有了这些发现，我假设PAQR8增益会促进乳腺癌转移和耐药性。 AIM 1地址 PAQR8收益是否促进转移。我将在原始移植物上均等的PAQR8-的混合物过表达和控制细胞进入免疫功能低下的小鼠，并跟踪转移性病变的形成通过体内荧光素酶成像。比较转移性和主要的两种不同细胞类型的比率病变，我希望与原发性相比，在转移中发现PAQR8过表达细胞的富集肿瘤。接下来，我将使用免疫荧光来检测被预测为PAQR8下游的活动介质原发性病变和转移性病变中的信号传导。 AIM 2解决PAQR8增益是否促进治疗反抗。我将执行无复发的生存分析，以确定是否注入了PAQR8-的小鼠治疗后比对照细胞注射的细胞更早地复发。调查残留和复发性肿瘤的组成，我将原位注入PAQR8过表达的混合物与AIM 1一样的对照细胞。原发性肿瘤将通过靶向治疗或化学疗法进行治疗，两者都将将分析残留病变和复发性肿瘤，以表达PAQR8过表达的细胞与对照的比率。我期望在残留肿瘤和复发性肿瘤中富含过表达PAQR8的细胞。最后，一个体外细胞竞争分析将确定PAQR8过表达是否介导了A中的治疗性孕酮剂量依赖性方式。提出的实验的结果将使人们对 PAQR8膜孕酮受体在乳腺癌转移和耐药性中的作用。