Aberrant Splice Variants as Potential Precision Biomarkers for Aggressive Prostate Cancers in African American Patients

异常剪接变异作为非洲裔美国患者侵袭性前列腺癌的潜在精确生物标志物

• 批准号: 10004669
• 负责人: Bi-Dar Wang
• 金额: $ 38.75万
• 依托单位: UNIVERSITY OF MARYLAND EASTERN SHORE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10004669
• 关键词: African American; Alternative Splicing; American; Automobile Driving; Biologic Characteristic; Biological; Biological Assay; Biological Markers; Biopsy Specimen; Breast; Cancer Etiology; Cancer cell line; Carcinoma; Catalytic Domain; Cell Line; Cell Proliferation; Cells; Cessation of life; Classification; Clinical; Colon; DU145; Data; Development; Discrimination; Disease; Disease Progression; Drug resistance; Environmental Risk Factor; European; Evaluation; Event; Exhibits; Expression Profiling; FGFR3 gene; Fluorescent in Situ Hybridization; Foundations; Future; Genes; Genomics; Goals; Grant; Hematologic Neoplasms; Human; Immunohistochemistry; In Vitro; Incidence; Individual; LNCaP; Lead; Length; Link; Malignant Neoplasms; Malignant neoplasm of prostate; Messenger RNA; MicroRNAs; Modeling; Molecular; Molecular Profiling; Neoplasm Metastasis; Neuroblastoma; Oligonucleotides; Oncogenic; Oncoproteins; PC3 cell line; Patients; Pharmaceutical Preparations; Phenotype; Phosphatidylinositol 4,5-Diphosphate; Phosphotransferases; Pilot Projects; Play; Population; Property; Prostate; Protein Isoforms; Proteins; RNA Splicing; Race; Recurrence; Research; Reverse Transcriptase Polymerase Chain Reaction; Role; Sampling; Sensitivity and Specificity; Series; Small Interfering RNA; Socioeconomic Factors; Socioeconomic Status; Solid; Solid Neoplasm; Specimen; Spliced Genes; Structure; TSC2 gene; Testing; Therapeutic; Time; Transcription Process; Transfection; Treatment Efficacy; Tumor Cell Line; United States; United States National Institutes of Health; VCaP; Validation; Variant; Western Blotting; advanced prostate cancer; cancer biomarkers; cancer diagnosis; cancer health disparity; cytotoxicity; design; docetaxel; ethnic difference; genetic element; genome-wide; inhibitor/antagonist; leukemia; leukemia/lymphoma; mRNA Precursor; men; migration; mortality; neoplastic cell; novel; novel marker; novel therapeutics; outcome forecast; overexpression; potential biomarker; precision medicine; prostate cancer cell; prostate cancer cell line; racial and ethnic; responders and non-responders; small molecule inhibitor; success; therapeutic target; tumor

PROJECT SUMMARY/ABSTRACT Prostate cancer (PCa) is now the most frequently diagnosed cancer and the second most common cause of cancer deaths in men in United States. Notably, African Americans (AAs) exhibited 1.6-fold higher incidence and 2.4-fold higher mortality rates compared to European Americans (EAs). Despite multiple socioeconomic factors postulated to explain the observed PCa disparities, higher recurrence and mortality rates remained even after adjustment of socioeconomic status in AAs, suggesting that intrinsic biological differences account for, at least, part of PCa disparities. Our previous genomic studies have revealed that intrinsic genomic differences do exist between AA and EA PCa. These genetic elements, including population-specific and -enriched microRNAs, mRNAs and alternative splicing variants, have been identified and were hypothesized to contribute to the differential PCa properties between AA and EA. In the pilot study, our preliminary results revealed ~2,500 differential splicing (DS) events occurring in AA and EA PCa. Among these DS genes, >70% of the genes were functionally linked to cancer diseases. These results lead to our hypothesis that aberrant mRNA splicing may play a critical but largely unknown role for driving the PCa disparities. Towards this hypothesis, we have performed RT-PCR validations and functional analyses of AA-enriched splice variants (such as PIK3CD, FGFR3, TSC2 and RASGRP2 splice variants) in PCa samples. Our preliminary results confirmed that these AA-enriched variants were highly expressed in AA PCa and contributed to more aggressive cancer phenotypes. In this proposal, we will focus on investigating the expression profiles of PIK3CD long and short splice variants (PIK3CD-L and PIK3CD-S) in a panel of PCa and other solid/hematologic tumor cell lines, and elucidating functional impacts of these splice variants in PCa aggressiveness and drug resistance. Guided by strong preliminary data, we propose to pursue three Specific Aims to character molecular functions of the PIK3CD splice variants underlying PCa aggressiveness: 1) Validate the expression profiles of PIK3CD splice variants in PCa specimens and cell lines derived from AA and EA patients; 2) Determine the functional roles of PIK3CD-S expression in disease aggressiveness and drug resistance in PCa and evaluate the feasibility of PIK3CD- S/PIK3CD-L ratios as potential biomarker; and 3) Screen for the effective therapeutic molecules (small molecule inhibitors, siRNAs and/or splice switching oligos) to reduce the cancer phenotypes in PIK3CD-S overexpressing cells. Collectively, our proposed research will broadly contribute to the field of cancer health disparities by characterizing the molecular signatures of aberrant splice variants in cancers (including PCa and other solid/hematologic tumors), and deciphering the molecular mechanisms of aberrant splicing underlying PCa aggressiveness (i.e. enhanced cell proliferation and invasion) and drug resistance. This proposal will allow us to further design for splice variant- specific siRNAs and screen for SMIs that can effectively inhibit the aggressive form of splice variants in PCa. These findings and efforts may facilitate the development of potential biomarkers and therapeutic strategy to treat aggressive PCa. Finally, the success of this SC1 proposal will set a strong foundation for the PI and his research team to further pursue a NIH competitive grant (i.e. R01 or R21) for future clinically-related studies.

项目总结/摘要 前列腺癌（PCa）是目前最常诊断的癌症， 美国男性癌症死亡的常见原因。非洲裔美国人（AAs） 与欧洲人相比， 美国人（EA）。尽管有多种社会经济因素可以解释观察到的 PCa差异，较高的复发率和死亡率即使在调整后仍然存在， 社会经济地位的AA，这表明内在的生物学差异占，在 至少，部分PCa差异。 我们先前的基因组研究已经揭示了内在的基因组差异确实存在于 AA和EA PCa。这些遗传因素，包括群体特异性和富集性 microRNA、mRNA和可变剪接变体，已经被鉴定出来， 假设有助于AA和EA之间的差异PCa属性。试点 我们的初步结果显示，AA中发生了约2，500个差异剪接（DS）事件。 EA PCa。在这些DS基因中，>70%的基因在功能上与癌症相关 疾病这些结果导致我们假设，异常的mRNA剪接可能发挥关键作用， 但在很大程度上未知的作用，推动PCa的差距。 针对这一假设，我们进行了RT-PCR验证和功能分析， AA富集的剪接变体（如PIK 3CD、FGFR 3、TSC 2和RASGRP 2剪接变体）， PCa样品。我们的初步结果证实，这些富含AA的变体是高度相关的。 在AA PCa中表达，并导致更具侵袭性的癌症表型。在这一提议中， 我们将集中研究PIK 3CD长和短剪接变体的表达谱， （PIK 3CD-L和PIK 3CD-S）在一组PCa和其他实体/血液肿瘤细胞系中的表达，和 阐明这些剪接变体在PCa侵袭性和药物治疗中的功能影响 阻力在强有力的初步数据的指导下，我们提出了三个具体目标， PIK 3CD剪接变体的特征分子功能是PCa侵袭性的基础：1） PIK 3CD剪接变体在PCa标本和细胞系中的表达谱 2）确定PIK 3 CD-S表达在AA和EA患者中的功能作用; PIK 3CD-1基因治疗前列腺癌的可行性研究 S/PIK 3CD-L比率作为潜在的生物标志物;和3）筛选有效的治疗分子 （小分子抑制剂、siRNA和/或剪接转换寡核苷酸）以减少癌症 PIK 3CD-S过表达细胞中的表型。 总的来说，我们提出的研究将为癌症健康领域做出广泛贡献。 通过表征癌症中异常剪接变异体的分子特征 （包括PCa和其他实体/血液肿瘤），并破译分子机制 异常剪接导致PCa侵袭性（即增强的细胞增殖和 侵袭）和耐药性。这一建议将使我们能够进一步设计剪接变体- 特异性siRNA和筛选可以有效抑制剪接侵袭性形式的SMI PCa中的变体。这些发现和努力可能有助于开发潜在的 生物标志物和治疗策略来治疗侵袭性PCa。最后，SC 1的成功 该提案将为PI及其研究团队进一步追求NIH奠定坚实的基础 竞争性补助金（即R 01或R21），用于未来的临床相关研究。