Epigenetic modulation of SEC24D and circulating miR-605 in renal cell carcinoma

肾细胞癌中 SEC24D 和循环 miR-605 的表观遗传调节

• 批准号: 10359448
• 负责人: Roberto Pili
• 金额: $ 6.87万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-25 至 2023-02-24
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10359448
• 关键词: Epigenetic; modulation; SEC24D; circulating; miR

One of the major challenges in the clinical management of clear cell renal cell carcinoma (ccRCC) is the acquired resistance to targeted therapies. Although microRNAs (miRNA) are potential predictors of cancer progression, to date there are no molecular studies reporting the role of circulating miRNA as biomarkers for predicting response to treatments in ccRCC. Our group has been conducting clinical trials in ccRCC patients with the aim to develop effective combination therapies. We have recently completed a Phase I/II clinical trial to assess the clinical benefit of the histone deacetylase inhibitor (HDACi) vorinostat in combination with the VEGF blocker bevacizumab in patients with prior tyrosine kinase inhibitors (TKIs) and assessed the expression of serum miRNA pre and post treatment. Exiqon microarray analysis has revealed the differential regulation of 134 microRNAs in the post- vs. pre-treatment samples. In particular, in patients who achieved an objective response (Responders) we have observed a significant up regulation of miRNA-605 at baseline as compared to patients who had progressive disease (Progressors). Interestingly, further analysis has revealed that following treatment downregulation of miR-605 expression occurred only in the Responders group. The decrease of miR-605 in the circulation suggests its availability inside the cells. Our bioinformatics prediction algorithms have revealed that SEC24D, a member of coat protein complex II (COPII), is targeted by miR-605. SEC24D is involved in the selection of cargo, concentration, and secretion of biomolecules such as proteins and RNA. To evaluate the effect of HDAC inhibition on SEC24D ccRCC cells were treated with vorinostat and assessed SEC24D expression. We found significant inhibition following HDACi treatment. Furthermore, we observed an increase in intracellular levels and decrease in exosomal miR-605 in 786-0 cells, suggesting an effect of vorinostat on miR-605 secretion. In addition, we also observed the inhibition of mutant p53 expression, as a reported target of miR-605. The current proposed study is aimed to determine a specific miRNA alteration associated with response to epigenetic modulators. Our hypothesis is that the inhibition of SEC24D leads to decrease of extracellular secretion of miR-605 which correlates with response to epigenetic modulators. We have designed the following specific aims to test our hypothesis: Specific Aim 1- To determine the effect of HDAC inhibition on SEC24D and miR-605 in ccRCC cells and patient derived tumor xenografts (PDXs); Specific Aim 2- To assess the correlation between clinical outcome and miR-605 expression in serum from RCC patients receiving HDAC inhibitors in combination with VEGF and PD1/PD-L1 inhibitors. The results from this pilot project will identify a specific circulating miRNA as potential biomarker of response to HDACi treatment that will need to be validated in further prospective clinical studies. Identification of specific molecular alterations of miR-605 and SEC24D in ccRCC patients will provide the rationale for future personalized clinical trials to evaluate the efficacy of HDACi in combination strategies.

肾透明细胞癌（ccRCC）临床管理的主要挑战之一是 对靶向治疗产生耐药性。尽管microRNA（miRNA）是癌症的潜在预测因子， 尽管如此，到目前为止，还没有分子研究报告循环miRNA作为生物标志物的作用， 预测对ccRCC治疗的反应。我们的团队一直在ccRCC患者中进行临床试验 目的是开发有效的联合疗法。我们最近完成了I/II期临床试验， 评估组蛋白去乙酰化酶抑制剂（HDACi）伏立诺他联合VEGF治疗的临床获益 受体阻滞剂贝伐珠单抗在既往接受过酪氨酸激酶抑制剂（TKI）治疗的患者中的应用，并评估了 治疗前和治疗后的血清miRNA。微阵列分析揭示了基因表达的差异调节， 治疗后与治疗前样本中有134种microRNA。特别是在达到目标的患者中， 我们已经观察到基线时miRNA-605的显著上调， 进行性疾病（Progressors）。有趣的是，进一步的分析显示， 治疗后，miR-605表达的下调仅发生在应答者组中。的 循环中miR-605的减少表明其在细胞内的可用性。我们的生物信息学预测 算法已经揭示，SEC 24 D，外壳蛋白复合物II（COPII）的成员，被miR-605靶向。 SEC 24 D参与货物的选择、浓缩和生物分子如蛋白质的分泌 和RNA。为了评估HDAC抑制对SEC 24 D的影响，将ccRCC细胞用伏立诺他处理， 评估SEC 24 D表达。我们发现HDACi治疗后有显著的抑制作用。而且我们 在786-0细胞中观察到细胞内水平的增加和外泌体miR-605的减少，表明 伏立诺他对miR-605分泌的影响。此外，我们还观察到突变型p53的抑制作用， 表达，作为miR-605的报告靶点。目前提出的研究旨在确定一个具体的 与表观遗传调节剂反应相关的miRNA改变我们的假设是 SEC 24 D导致miR-605的细胞外分泌减少，这与表观遗传学应答相关。 调制器。我们设计了以下具体目标来验证我们的假设：具体目标1-为了 确定HDAC抑制对ccRCC细胞和患者来源的肿瘤中SEC 24 D和miR-605的影响 异种移植物（PDX）;具体目标2-评估临床结果与miR-605之间的相关性 来自接受HDAC抑制剂与VEGF和PD 1/PD-L1组合的RCC患者的血清中的表达 抑制剂的该试验项目的结果将确定一种特定的循环miRNA作为潜在的生物标志物， 对HDACi治疗的反应需要在进一步的前瞻性临床研究中验证。识别 在ccRCC患者中miR-605和SEC 24 D的特异性分子改变将为未来的研究提供依据。 个性化临床试验，以评估HDACi在联合策略中的疗效。