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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要前列腺癌（PCa）现在是最常诊断的癌症，也是第二大癌症美国男性癌症死亡的常见原因。值得注意的是，非裔美国人 (AA) 与欧洲相比，发病率高出 1.6 倍，死亡率高出 2.4 倍美国人（EA）。尽管假设有多种社会经济因素来解释观察到的情况即使在调整后，PCa 差异、较高的复发率和死亡率仍然存在 AA 的社会经济地位，表明内在的生物学差异在至少，这是 PCa 差异的一部分。我们之前的基因组研究表明，内在的基因组差异确实存在于 AA 和 EA PCa。这些遗传元素，包括特定人群和丰富的 microRNA、mRNA 和选择性剪接变体已被鉴定并被假设导致 AA 和 EA 之间的 PCa 特性差异。在试点中研究中，我们的初步结果显示 AA 中发生了约 2,500 个差异剪接 (DS) 事件和 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对于导致 PCa 差异的作用很大程度上未知。针对这一假设，我们进行了 RT-PCR 验证和功能分析富含 AA 的剪接变体（例如 PIK3CD、FGFR3、TSC2 和 RASGRP2 剪接变体） PCa 样本。我们的初步结果证实，这些富含 AA 的变体高度在 AA PCa 中表达并导致更具侵袭性的癌症表型。在这个提案中，我们将重点研究 PIK3CD 长剪接变体和短剪接变体的表达谱 (PIK3CD-L 和 PIK3CD-S) 在一组 PCa 和其他实体/血液肿瘤细胞系中，以及阐明这些剪接变异对 PCa 侵袭性和药物的功能影响反抗。在强有力的初步数据的指导下，我们建议追求三个具体目标 PCa 侵袭性背后的 PIK3CD 剪接变体的特征分子功能：1) 验证 PCa 标本和细胞系中 PIK3CD 剪接变体的表达谱源自 AA 和 EA 患者； 2)确定PIK3CD-S表达的功能作用 PCa 的疾病侵袭性和耐药性并评估 PIK3CD- 的可行性 S/PIK3CD-L 比率作为潜在的生物标志物； 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 PIK3CD-S 过表达细胞的表型。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体的分子特征来确定差异（包括前列腺癌和其他实体/血液肿瘤），并破译分子机制 PCa 侵袭性背后的异常剪接（即增强的细胞增殖和侵袭）和耐药性。该提案将使我们能够进一步设计剪接变体- 特异性 siRNA 并筛选可有效抑制攻击性剪接形式的 SMI PCa 的变体。这些发现和努力可能会促进潜在的发展治疗侵袭性 PCa 的生物标志物和治疗策略。终于，这次SC1的成功该提案将为 PI 及其研究团队进一步追求 NIH 奠定坚实的基础用于未来临床相关研究的竞争性资助（即 R01 或 R21）。