PO.PR02.01 · 预防研究

用于癌症预防与阻断的天然产物发现的靶点验证及高通量筛选检测法开发

Target validation and high-throughput screening assay development for natural product discovery for cancer prevention and interception

编号 942 展板 1 时间 4/19 02:00–05:00 区域 Section 37 主讲 Yurong Song, PhD

分会场 Experimental Chemoprevention and Interception: Data and Tools

该海报暂无可下载的资料 AACR 官方页面

作者与单位 Authors & Affiliations

Yurong Song¹, Brandon Somerville¹, Kajal Biswas², Karim Baktiar¹, Liankun Song³, Sara Sanders⁴, Tanja Grkovic⁵, Ligia A. Pinto¹, Ana Catarina Menezes⁶, Paul P. Liu⁶, Matthew J. Hart⁷, Chinthalapally V. Rao⁷, Shugeng Cao⁸, Xin Chen⁸, Xu Wu⁹, Xiaolin Zi³, Mark J. Henderson⁴, Barry R. O'Keefe⁵, Altaf Mohammed², Robert H. Shoemaker²

¹Frederick National Laboratory for Cancer Research, Frederick, MD,²Division of Cancer Prevention, National Cancer Institute, Bethesda, MD,³Department of Urology, University of California, Irvine, CA,⁴National Center for Advancing Translational Science, Rockville, MD,⁵National Cancer Institute at Frederick, Frederick, MD,⁶National Human Genome Research Institute, Bethesda, MD,⁷University of Oklahoma Health Sciences Center, Oklahoma City, OK,⁸University of Hawai'i at Hilo, Hilo, HI,⁹Massachusetts General Hospital and Harvard Medical School, Charlestown, MA

摘要 Abstract

中文摘要

背景：天然产物（NPs）是具有癌症预防/阻断潜力的生物活性化合物的丰富来源。NPs提供独特的化学多样性和安全的人类使用历史，使其成为长期使用的有吸引力的候选物。高通量筛选和机制研究是发现有前景的先导化合物并将其转化为有效且安全的癌症预防和阻断策略的关键。为支持“天然产物用于癌症阻断与预防的发现与开发（DDNP-CIP）”计划，我们旨在验证具有潜在临床相关性的分子靶点（SKP2、TEAD2、5-LOX和RUNX1），并建立和优化检测法，以支持用于癌症预防的高通量NPs文库筛选。方法：使用组织微阵列进行靶点验证。SKP2是SKP2-SCF E3连接酶复合物的组成部分，TEAD2是DNA结合转录因子，分别在前列腺癌和肝癌的TMA中评估其表达水平。此外，开发或改进了基于细胞的平台，以鉴定表现出免疫调节活性或靶向5-LOX（一种脂质过氧化酶）或RUNX1（一种对造血分化至关重要且在血液恶性肿瘤中频繁突变的转录因子）的NPs。这些检测法利用了稳定表达的报告基因细胞系。结果：与正常组织相比，SKP2在68.6%的前列腺增生、97.6%的前列腺上皮内瘤变和81.9%的腺癌中显著过表达。TEAD2在肝增生中高表达，并在肝炎（p=0.0041）、肝细胞癌（p=0.0012）和肝内胆管癌（p<0.0001）中显著上调。为鉴定具有免疫调节活性的NPs，改进并优化了使用THP-1 ISRE FRET报告基因细胞系的检测法。测试了参考化合物和初始筛选板，若干样本引发了阳性反应。生成并验证了表达5-LOX的细胞系。使用阳性化合物去甲二氢愈创木酸（nordihydroguaiaretic acid）的抑制检测显示，在花生四烯酸处理下呈剂量依赖性抑制（0.1μM时29%至1μM时92%）且无毒性。为筛选具有剪接调节活性的NPs，生成了带有在家族性血小板疾病伴髓系恶性肿瘤患者中鉴定出的RUNX1突变的纳米荧光素酶报告基因细胞系。若干参考化合物在选定的突变中显示出阳性反应。小结：经过验证的靶点和检测平台为高通量NPs筛选奠定了基础。这些工作推进了DDNP-CIP鉴定和开发用于癌症预防与阻断的NPs的使命。部分由国家癌症研究所资助，合同编号HHSN261201500003I。

查看英文原文 English abstract

Background Natural products (NPs) represent a rich source of bioactive compounds with cancer prevention/interception potential. NPs offer unique chemical diversity and a history of safe human use, making them attractive candidates for long-term use. High-throughput screening and mechanistic studies are key to discovering promising leads and translating them into effective and safe cancer prevention and interception strategies. To support the Discovery and Development of Natural Products for Cancer Interception and Prevention (DDNP-CIP) Initiative, we aimed to validate potentially clinically relevant molecular targets (SKP2, TEAD2, 5-LOX and RUNX1) and establish and optimize assays to enable high-throughput NPs library screening for cancer prevention. Methods Target validation was performed using tissue microarrays. SKP2, a component of the SKP2-SCF E3 ligase complex, and TEAD2, a DNA-binding transcription factor, were evaluated for their expression levels in TMAs of prostate and liver cancer, respectively. In addition, cell-based platforms were developed or adapted to identify NPs exhibiting immune-modulating activity or targeting either 5-LOX, a lipid-peroxidizing enzyme, or RUNX1, a transcription factor critical for hematopoietic differentiation and frequently mutated in hematologic malignancies. These assays utilized stably expressing reporter cell lines. Results SKP2 was significantly over expressed in 68.6% of prostate hyperplasia, 97.6% of prostate intraepithelial neoplasia, and 81.9% of adenocarcinoma compared with normal tissue. TEAD2 was highly expressed in liver hyperplasia and significantly upregulated in hepatitis (p = 0.0041), hepatocellular carcinoma (p = 0.0012), and intrahepatic cholangiocarcinoma (p < 0.0001). To identify NPs with immune-modulating activity, an assay using the THP-1 ISRE FRET reporter cell line was adapted and optimized. Reference compounds and initial challenge plates were tested, with several samples eliciting positive responses. A cell line expressing 5-LOX was generated and validated. An inhibition assay using a positive compound nordihydroguaiaretic acid demonstrated dose-dependent inhibition (29% at 0.1uM to 92% at 1uM) upon arachidonic acid treatment without toxicity. To screen NPs for splicing modulating activity, nano-luciferase reporter cell lines with RUNX1 mutations identified in patients with familial platelet disorder with associated myeloid malignancy were generated. Several reference compounds showed positive responses in selected mutations. Summary Validated targets and assay platforms establish a foundation for high-throughput NPs screening. These efforts advance the DDNP-CIP's mission to identify and develop NPs for cancer prevention and interception. Funded partly by the National Cancer Institute under Contract No. HHSN261201500003I

利益披露 Disclosure

Y. Song, None.. B. Somerville, None.. K. Biswas, None.. K. Baktiar, None.. L. Song, None.. S. Sanders, None.. T. Grkovic, None.. L. A. Pinto, None.. A. Menezes, None.. P. P. Liu, None.. M. J. Hart, None.. C. V. Rao, None.. S. Cao, None.. X. Chen, None.. X. Wu, None.. X. Zi, None.. M. J. Henderson, None.. B. R. O'Keefe, None.. A. Mohammed, None.. R. H. Shoemaker, None.

← 返回 AACR 2026 检索