Aptamer-siRNA gene knockdown to treat epithelial triple negative breast cancer

适体-siRNA 基因敲除治疗上皮性三阴性乳腺癌

基本信息

批准号：
9766827
负责人：
Judy Lieberman
金额：
$ 39.27万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-24 至 2020-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9766827
关键词：
Affect Affinity Amyloidosis Antibodies Binding Body Fluids Breast Cancer Cell Breast Cancer Genetics Breast Cancer Model Breast Cancer cell line Breast Epithelial Cells Bypass Cancer Prognosis Carcinoma Cell Proliferation Cell membrane Cells Chimera organism Cleaved cell Clinical Clinical Trials Cytoplasm Cytotoxic Chemotherapy Dependence Disease Disseminated Malignant Neoplasm Drug Kinetics Drug resistance Encapsulated Endosomes Epithelial Epithelial Cells Epithelium Estrogens Fibroblasts Gene Expression Gene Silencing Gene Targeting Genes Genetic Goals Hepatocyte Human In Vitro Inflammation Injections Knowledge Link Liposomes Liver Lung Malignant - descriptor Malignant Neoplasms Mammary Gland Parenchyma Mammary Neoplasms Mammospheres Maximum Tolerated Dose Mesenchymal Methods Mitosis Mus Natural Immunity Normal Cell PLK1 gene Pharmaceutical Preparations Pharmacodynamics Phase Progesterone RNA RNA Interference RNA interference screen RNA-Binding Proteins Relapse Serum Small Interfering RNA Small RNA Stains Structure TACSTD1 gene Testing Therapeutic Tissues Toxic effect Tumor Antigens Tumor Initiators Tumor Suppression Xenograft Model Xenograft procedure animal imaging antitumor effect aptamer base cancer biomarkers cancer cell cancer stem cell cancer subtypes cancer therapy design drug candidate drug relapse flexibility genome-wide in vitro Assay in vivo knock-down malignant breast neoplasm nanoparticle delivery novel nucleic acid structure orthotopic breast cancer outcome forecast pharmacokinetics and pharmacodynamics phase 3 study preclinical study programs public health relevance receptor research clinical testing self-renewal subcutaneous targeted treatment triple-negative invasive breast carcinoma tumor tumor initiation uptake virtual

项目摘要

DESCRIPTION (provided by applicant): RNA interference (RNAi) offers the exciting opportunity to treat disease by knocking down disease-causing genes. Recent early phase clinical trials have shown promising and sustained gene knockdown and/or clinical benefit in a handful of diseases caused by aberrant gene expression in the liver. The major obstacle to harnessing RNAi for cancer treatment is delivery of small RNAs to disseminated cancer cells. Most epithelial cancer cells and the tumor-initiating cells (T-IC) within them highly express EpCAM, the first described tumor antigen. All epithelial breast cancer cell lines we tested stain brightly for EpCAM, while immortalized normal breast epithelial cells and fibroblasts do not. Targeted gene knockdown in epithelial cancer cells in vitro can be achieved using chimeric RNAs composed of a structured RNA, called an aptamer, selected for high affinity binding to EpCAM, that is covalently linked to an siRNA. These EpCAM aptamer-siRNA chimeras (AsiC) are taken up by EpCAM+ cells and selectively cause gene knockdown in epithelial breast cancer cells, but not normal epithelial cells. Moreover knockdown of PLK1 with EpCAM-AsiCs suppresses colony and mammosphere formation of epithelial breast cancer lines, in vitro assays of tumor-initiating potential, and tumor initiation. Subcutaneously injected PLK1 EpCAM-AsiCs are taken up specifically by EpCAM+ basal-A triple negative breast cancer (TNBC) orthotopic xenografts and cause rapid tumor regression. TNBC has the worst prognosis of any breast cancer and there is no targeted therapy for it. The goal of this proposal is to evaluate whether EpCAM-AsiCs can be used for targeted gene knockdown to treat epithelial (basal-like) TNBC cancers, sparing normal cells, and eliminate the T-ICs within them. One goal is to define which breast cancer subtypes can be targeted by EpCAM-AsiCs and determine how EpCAM level affects uptake and gene silencing. Relative uptake/knockdown in cancer cells expressing EpCAM and normal epithelium will be evaluated in human breast cancer tissue explants. Another goal is to determine whether EpCAM-AsiCs can target breast T-ICs to disrupt tumor initiation. An important goal is to optimize the drug-like features of EpCAM-AsiCs to produce a drug candidate suitable for clinical evaluation. To achieve this aim, we will optimize EpCAM-AsiCs for cell uptake, endosomal release, systemic delivery and in vivo gene knockdown. Pharmacokinetics (PK) and pharmacodynamics (PD) of EpCAM-AsiC uptake and gene silencing and tumor suppression will be evaluated using live animal imaging in TNBC cell line xenograft models. As proof of principle, we will evaluate the antitumor effect of knockdown of PLK1, which is needed for cell proliferation. In addition knockdown of novel gene targets identified in a genome-wide siRNA screen for TNBC genetic dependencies will be evaluated in mouse xenograft models. At the end of this program, we will have an optimized EpCAM-AsiC and knowledge of its PK, PD and possible toxicity, to lay the groundwork for formal toxicity and other preclinical studies needed to initiate clinical proof of concept studies.

描述（由申请人提供）：RNA 干扰 (RNAi) 提供了通过敲除致病基因来治疗疾病的令人兴奋的机会。最近的早期临床试验表明，基因敲除和/或对少数引起的疾病有希望和持续的临床益处。利用 RNAi 治疗癌症的主要障碍是将小 RNA 递送至大多数上皮癌细胞及其内的肿瘤起始细胞 (T-IC)。表达 EpCAM，这是第一个描述的肿瘤抗原。我们测试的所有上皮乳腺癌细胞系都对 EpCAM 进行了明亮的染色，而永生化的正常乳腺上皮细胞和成纤维细胞则不能使用由组成的嵌合 RNA 来实现体外上皮癌细胞中的靶向基因敲除。一种称为适体的结构化 RNA，选择与 EpCAM 高亲和力结合，与这些 EpCAM 适体-siRNA 共价连接。嵌合体 (AsiC) 被 EpCAM+ 细胞摄取，并选择性地导致上皮性乳腺癌细胞中的基因敲低，但不会导致正常上皮细胞的基因敲低。此外，体外试验显示，EpCAM-AsiCs 敲低 PLK1 会抑制上皮性乳腺癌细胞系的集落和乳腺球形成。皮下注射的 PLK1 EpCAM-AsiC 会被 EpCAM+ 特异性吸收。 basal-A 三阴性乳腺癌（TNBC）原位异种移植并导致肿瘤快速消退，TNBC 是所有乳腺癌中预后最差的，并且没有针对它的靶向治疗。该提案的目的是评估 EpCAM-AsiC 是否可以用于治疗。用于靶向基因敲除来治疗上皮（基底细胞样）TNBC 癌症，保护正常细胞并消除其中的 T-IC，其中一个目标是确定可以靶向哪些乳腺癌亚型。另一个目标是确定 EpCAM-AsiCs 是否可以靶向乳腺 T，并确定 EpCAM 水平如何影响表达 EpCAM 和正常上皮的癌细胞的相对摄取/敲低。 -扰乱肿瘤发生的 IC 的一个重要目标是优化 EpCAM-AsiC 的类药物特性，以产生适合临床评估的候选药物来实现这一目标。我们将优化 EpCAM-AsiC 的细胞摄取、内体释放、全身递送和体内基因敲除，并使用活体动物成像评估 EpCAM-AsiC 摄取的药代动力学 (PK) 和药效学 (PD) 以及基因沉默和肿瘤抑制。在 TNBC 细胞系异种移植模型中，作为原理证明，我们将评估细胞增殖所需的 PLK1 敲除的抗肿瘤作用以及新基因靶标的敲除。在全基因组 siRNA 筛选中鉴定的 TNBC 遗传依赖性将在小鼠异种移植模型中进行评估。在该计划结束时，我们将获得优化的 EpCAM-AsiC 及其 PK、PD 和可能的毒性的知识，从而奠定基础。用于启动临床概念验证研究所需的正式毒性和其他临床前研究。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Tapping the RNA world for therapeutics.

利用 RNA 世界进行治疗。

DOI：
发表时间：
2018
期刊：
影响因子：
16.8
作者：
Lieberman; Judy
通讯作者：
Judy

Gene Knockdown by EpCAM Aptamer-siRNA Chimeras Suppresses Epithelial Breast Cancers and Their Tumor-Initiating Cells.

EpCAM 适体-siRNA 嵌合体的基因敲除可抑制上皮性乳腺癌及其肿瘤起始细胞。

DOI：
发表时间：
2015-10
期刊：
影响因子：
5.7
作者：
Gilboa;Hamar, Peter;Le, Minh T N;Wheeler, Lee Adam;Trifonova, Radiana;Petrocca, Fabio;Wittrup, Anders;Lieberman, Judy
通讯作者：
Lieberman, Judy

Blocking the recruitment of naive CD4+ T cells reverses immunosuppression in breast cancer.

阻断初始 CD4 T 细胞的募集可逆转乳腺癌的免疫抑制。

DOI：
发表时间：
2017-04
期刊：
影响因子：
44.1
作者：
Su, Shicheng;Liao, Jianyou;Liu, Jiang;Huang, Di;He, Chonghua;Chen, Fei;Yang, LinBing;Wu, Wei;Chen, Jianing;Lin, Ling;Zeng, Yunjie;Ouyang, Nengtai;Cui, Xiuying;Yao, Herui;Su, Fengxi;Huang, Jian;Lieberman, Judy;Liu, Qiang;Song, Erwei
通讯作者：
Song, Erwei