Consequences and mechanism of aberrant splicing in African American prostate cancer disparities

非裔美国人前列腺癌差异中异常剪接的后果和机制

基本信息

批准号：
9884534
负责人：
Mariano A. Garcia-Blanco
金额：
$ 35.74万
依托单位：
GEORGE WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-03-01 至 2022-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9884534
关键词：
1-Phosphatidylinositol 3-Kinase Address African American Alternative Splicing American Automobile Driving Back Binding Biological Assay CRISPR/Cas technology Catalytic Domain Cell Line Cell Proliferation Cell-Free System Cells Characteristics Clip Code Complement Complementary DNA Computer Analysis DNA Methylation DNA cassette Data Development Disease Elements Enhancers European Event Exhibits Exons FGFR3 gene Fibroblast Growth Factor Receptors Future Generations Genes Health Histones Human In Vitro Incidence Individual Integrins Knock-out Malignant Neoplasms Malignant neoplasm of prostate Mediating Messenger RNA MicroRNAs Molecular Mus Nature Neoplasm Metastasis Oncogenic Patients Pattern Phenotype Phosphatidylinositol 4,5-Diphosphate Phosphotransferases Population Production Protein Isoforms Proteins Publishing RNA Sequences RNA Splicing Race Receptor Protein-Tyrosine Kinases Recombinants Regulation Regulator Genes Resistance Role SRSF2 gene Signal Pathway Signal Transduction Site Specimen System Techniques Technology Testing Transcriptional Activation Transplantation Tumor Suppressor Genes Ubiquitination Up-Regulation Variant Vascular Endothelial Growth Factors Xenograft Model Xenograft procedure cancer health disparity cancer survival cellular imaging differential expression druggable target epithelial to mesenchymal transition exon skipping experimental study fibroblast growth factor-14 gain of function genetically modified cells health disparity high risk imaging approach in vitro Assay knock-down mimetics mortality mouse model outcome forecast overexpression prostate cancer cell prostate cancer cell line prostate cancer risk racial disparity small hairpin RNA small molecule small molecule inhibitor stable cell line statistics survival outcome targeted treatment transcription factor tumor tumor growth

项目摘要

Summary There are striking population (race) disparities in prostate cancer (PCa) risk and survival outcome borne out of current health statistics. This is particularly evident between African American (AA) patients and their European American (EA) counterparts, where AAs exhibit a 1.5 to 2 fold higher risk of PCa incidence and mortality. We demonstrate that differential alternative RNA splicing takes place for oncogenes and tumor suppressor genes in PCa specimens of AA compared to EA patients. The differential splicing events result in the enrichment and in many instances generation of AA-specific splice variants not observed in EA cancers. We have cloned the AA and EA splice variant cDNAs for both phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit delta (PIK3CD) and fibroblast growth factor receptor 3 (FGFR3), and ectopically introduced the clones individually into PCa cell lines. Compared to the EA variant-expressing lines, the AA variant-expressing lines are more oncogenic in in vitro cell proliferation and invasion assays, and exhibit greater tumor growth/metastasis and resistance to small molecule targeted therapy in xenograft mouse models. These findings may explain, in part, the observed cancer health disparities in the AA population relative to other racial groups. The mechanism responsible for the formation of AA-specific or -enriched mRNA splice variants are hypothesized to be due to the dysregulated expression of 6 key splicing factor s in AA PCa specimens. We refer to this dysregulation and associated production of AA-specific splice variants as an “AA splicing factor code in PCa disparities.” Notwithstanding these developments, a number of questions remain unresolved. First, what is the mechanism(s) of dysregulated expression of the 6 splicing factors? Second, can the production of AA-specific splice variants via exon skipping be directly attributed to the 6 dysregulated splicing factors? Lastly, how do the encoded proteins of the AA-specific short variants of PIK3CD and FGFR3 mediate a more oncogenic phenotype in PCa cells? The objectives of this application are to address these questions, and to provide a mechanistic framework of population-related differences in alternative RNA splicing, which is anticipated to facilitate future identification of druggable targets (e.g. splicing factors and/or resulting splice variant proteins) of AA tumor phenotype and disparities.

概括前列腺癌 (PCa) 风险和生存结果存在显着的人群（种族）差异当前的健康统计数据在非裔美国人 (AA) 患者和欧洲患者之间尤其明显。美国 (EA) 艾滋病，其中 AA 的 PCa 发病率和死亡率风险高出 1.5 至 2 倍。证明癌基因和抑癌基因发生不同的选择性 RNA 剪接与 EA 患者相比，AA 患者的 PCa 样本中差异剪接事件导致富集和在许多情况下，我们克隆了在 EA 癌症中未观察到的 AA 特异性剪接变体。磷脂酰肌醇-4,5-二磷酸 3-激酶催化亚基 Delta 的 AA 和 EA 剪接变体 cDNA (PIK3CD) 和成纤维细胞生长因子受体 3 (FGFR3)，并分别异位引入克隆与 EA 变体表达细胞系相比，AA 变体表达细胞系更多。致癌的体外细胞增殖和侵袭测定，并表现出更大的肿瘤生长/转移和这些发现可以部分解释：观察到 AA 人群相对于其他种族群体的癌症健康差异。负责形成 AA 特异性或富集 mRNA 剪接变体的原因被追踪为 AA PCa 标本中 6 个关键剪接因子的表达失调我们将这种失调称为 AA 特异性剪接变体的相关生产作为“PCa 差异中的 AA 剪接因子代码”。尽管取得了这些进展，但仍有一些问题尚未解决，首先，什么是。 6 种剪接因子表达失调的机制外显子跳跃导致的剪接变异直接归因于 6 个失调的剪接因子？ PIK3CD 和 FGFR3 AA 特异性短变体的编码蛋白介导更具致癌性 PCa 细胞的表型？本应用的目的是解决这些问题，并提供一个替代RNA剪接中群体相关差异的机制框架，预计将促进未来可药物靶标的识别（例如剪接因子和/或产生的剪接变体蛋白） AA 肿瘤表型和差异。