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

• 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
• 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

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）风险和生存结果中存在惊人的人口（种族）差异 当前的健康统计。这是非裔美国人（AA）患者及其欧洲人之间的尤其证据 American（EA）对应物，AAS表现出1.5至2倍的PCA事件和死亡率的风险更高。我们 证明对癌基因和肿瘤抑制基因进行差异替代RNA剪接 与EA患者相比，在AA的PCA标本中。差异剪接事件导致富集和 在许多情况下，在EA癌中未观察到AA特异性剪接变体。我们克隆了 磷脂酰肌醇-4,5-双磷酸3-激酶催化亚基三角洲的AA和EA剪接变体cDNA （PIK3CD）和成纤维细胞生长因子受体3（FGFR3），并分别引入克隆 进入PCA细胞系。与表达EA变异线相比，表达AA变体的线更多 体外细胞增殖和侵袭测定中的致癌性，暴露于更大的肿瘤生长/转移和 在Xenographotic小鼠模型中对小分子靶向治疗的抗性。这些发现可能部分解释了 相对于其他种族群体，AA人群中观察到的癌症健康差异。机制 负责AA特异性或富集的mRNA剪接变体的形成是由于 AA PCA标本中6个关键剪接因子S的表达失调。我们指的是这种失调和 相关的AA特异性剪接变体作为“ PCA分布中的AA剪接因子代码”的生产。 尽管有这些发展，但许多问题仍未解决。首先，什么是 6个剪接因子表达失调的机制？其次，可以生产AA特定的 通过跳过外显子跳的剪接变体直接归因于6个失调的剪接因子？最后，如何 PIK3CD和FGFR3的AA特异性短变体的编码蛋白质介导了更致癌的 PCA细胞中的表型？此应用程序的目标是解决这些问题，并提供 替代RNA剪接中与人口相关差异的机理框架，预计将 促进未来对可药靶标的识别（例如，剪接因子和/或由此产生的剪接变异蛋白） AA肿瘤表型和差异。