Nanotechnology for targeted therapy and fundamental understanding oftherapeutic resistance in triple negative breast cancer

用于靶向治疗的纳米技术和对三阴性乳腺癌治疗耐药性的基本了解

基本信息

批准号：
10593921
负责人：
Xiaoming He
金额：
$ 44.19万
依托单位：
UNIV OF MARYLAND, COLLEGE PARK
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10593921
关键词：
3-Dimensional American Animals Biological Availability Biomimetics Blood Blood Circulation Blood Proteins Body Weight Breast Cancer Cell Breast Cancer Model Breast Cancer Patient CD44 gene CRISPR/Cas technology Cancer Etiology Catalytic Domain Cell Survival Cell division Cells Cessation of life Chemotherapy-Oncologic Procedure Clinic Clinical Colorectal Cancer Complex Cytosol Data Defect Development Disease Drug resistance Encapsulated Endothelial Cells Essential Genes Foundations Gene Delivery Gene Dosage Genetic Transcription Genomics Growth Human Hyaluronic Acid Hydrophobicity Immune response In Vitro Lasers Ligands Lysosomes Malignant - descriptor Malignant Neoplasms Malignant neoplasm of prostate Mammalian Cell Mediating Messenger RNA Metformin MicroRNAs Microfluidics Microtubule Stabilization Microtubules Mutation Nanotechnology Nature Neoplasm Metastasis P-Selectin POLR2A gene Paclitaxel Patients Permeability Pharmaceutical Preparations Probability Proliferating Public Health Publishing RNA RNA Interference RNA Polymerase II RNA Stability RNA delivery Resistance Signal Transduction Small Interfering RNA TP53 gene Testing Therapeutic Translating Tumor Suppressor Genes Tumor Suppressor Proteins Variant Woman Work Xenograft procedure anti-cancer cancer cell cancer drug resistance cancer therapy cancer type cellular transduction chemotherapy cytokine drug relapse experience fucoidan human model hydrophilicity improved in vivo liposomal delivery liver function malignant breast neoplasm miniaturize mortality mouse model nanobomb nanoparticle new therapeutic target novel novel strategies novel therapeutics overexpression resistance mechanism small hairpin RNA stem cells stem-like cell systemic toxicity targeted treatment taxane therapeutic RNA therapy resistant three dimensional cell culture triple-negative invasive breast carcinoma tumor tumor growth tumor initiation tumorigenic uptake virtual

项目摘要

Project Summary/Abstract Breast cancer is the second leading cause of cancer-related deaths of American women. In particular, no targeted therapy is clinically available for nearly all triple negative breast cancer (TNBC). Cancer arises as a result of accumulating genetic alterations. Therefore, developing novel strategies to precisely target the genetic alterations of TNBC may be valuable for combating the malignant disease. TP53 is a pivotal tumor suppressor gene inactivated by mutation or deletion in most human cancers. Tremendous effort has been made to restore the activity of the p53 protein encoded by TP53 for cancer treatment. Unfortunately, no p53-based therapy has been successfully translated into the clinic, due to the complexity of p53 signaling. Therefore, identifying vulnerabilities conferred by TP53 deletion instead of restoring the p53 activity is a novel strategy for combating cancer. In our recent work published in Nature and Nature Nanotechnology, we revealed genomic deletion of TP53 is often accompanied by hemizygous (i.e., partial) loss of a neighboring gene POLR2A essential for cell survival, and virtually all 53% TNBCs with TP53 deletion harbor hemizygous POLR2A loss (TP53/POLR2Aloss). Our preliminary data show that suppressing POLR2A expression by RNA interference with small interfering RNA (siRNA) delivered using a low pH-activated nanobomb selectively inhibits the proliferation, survival, and tumorigenic potential of TP53/POLR2Aloss TNBC cells. The nanobomb protects the siRNA in blood and enables endo/lysosomal escape of the siRNA into the cytosol where the siRNA performs its POLR2A inhibition function after cell uptake. Moreover, the nanobomb-mediated delivery of POLR2A-targeting siRNA selectively inhibits the growth of orthotopic TP53/POLR2Aloss TNBC tumors, with no evident systemic toxicity demonstrated by the data on animal body weight and blood proteins (for liver function) and cytokines (for immune responses). However, a small fraction of breast cancer cells overexpressed with the variant CD44 (note: not the non- variant or normal CD44 on normal stem cells) have been shown to be particularly resistant to clinically used chemotherapy drugs of TNBC such as paclitaxel (PTX). Since POLR2A is indispensable for cancer cells to survive, we hypothesize that targeted co-delivery of the POLR2A-targeting siRNA and PTX to the variant CD44+ cancer cells can overcome the TNBC drug resistance. We will further develop the aforementioned low pH-activated nanobomb that has no active targeting, to be capable of actively targeting both the variant CD44+ cells and tumor vasculature. Since cancer metastasis is the major cause of cancer-related death, we will test the hypothesis using not only the aforementioned orthotopic/primary TNBC tumors but also metastatic TNBC model. Furthermore, we will investigate the mechanisms of resistance to the POLR2A-targeted therapy using not only 2D and xenograft but also 3D TNBC models generated using microfluidic approach developed by us. Collectively, this project may result in a novel therapy for drug-resistant TNBC with mechanistic understanding, which is invaluable for combating TNBC and possibly many other types of cancers harboring TP53 deletion.

项目概要/摘要乳腺癌是美国女性癌症相关死亡的第二大原因。特别是，没有临床上几乎所有三阴性乳腺癌（TNBC）都可以进行靶向治疗。癌症的出现是由于基因改变累积的结果。因此，开发新策略来精确靶向遗传 TNBC 的改变对于对抗这种恶性疾病可能有价值。 TP53是一种关键的肿瘤抑制因子在大多数人类癌症中，该基因因突变或缺失而失活。已付出巨大努力来恢复 TP53 编码的 p53 蛋白在癌症治疗中的活性。不幸的是，目前尚无基于 p53 的疗法由于p53信号传导的复杂性，已成功转化为临床。因此，识别 TP53 删除而不是恢复 p53 活性所带来的漏洞是一种新的对抗策略癌症。在我们最近发表在《自然》和《自然纳米技术》上的工作中，我们揭示了基因组缺失 TP53 通常伴随着细胞必需的邻近基因 POLR2A 的半合子（即部分）丢失生存率，几乎所有 53% 具有 TP53 缺失的 TNBC 都存在半合子 POLR2A 缺失 (TP53/POLR2Aloss)。我们的初步数据表明，通过小干扰RNA干扰抑制POLR2A表达使用低 pH 激活纳米炸弹递送的 RNA (siRNA) 可选择性抑制增殖、存活和 TP53/POLR2Aloss TNBC 细胞的致瘤潜力。纳米炸弹可以保护血液中的 siRNA，并使 siRNA 内/溶酶体逃逸到细胞质中，其中 siRNA 发挥其 POLR2A 抑制功能细胞摄取后。此外，纳米炸弹介导的 POLR2A 靶向 siRNA 选择性抑制原位 TP53/POLR2Aloss TNBC 肿瘤的生长，没有明显的全身毒性有关动物体重和血液蛋白（用于肝功能）和细胞因子（用于免疫反应）的数据。然而，一小部分乳腺癌细胞过度表达变体 CD44（注意：不是非正常干细胞上的变异或正常 CD44）已被证明对临床使用特别耐药 TNBC的化疗药物如紫杉醇(PTX)。由于 POLR2A 对于癌细胞的生存是不可或缺的存活下来，我们假设 POLR2A 靶向 siRNA 和 PTX 共同递送至变体 CD44+癌细胞可以克服TNBC耐药性。我们将进一步开发上述低 pH 激活纳米炸弹没有主动靶向，能够主动靶向变体 CD44+ 细胞和肿瘤血管系统。由于癌症转移是癌症相关死亡的主要原因，我们将测试该假设不仅使用上述原位/原发性 TNBC 肿瘤，还使用转移性 TNBC 模型。此外，我们将使用 POLR2A 靶向治疗来研究耐药机制不仅有 2D 和异种移植，还有使用我们开发的微流体方法生成的 3D TNBC 模型。总的来说，该项目可能会产生一种具有机制理解的耐药 TNBC 的新疗法，这对于对抗 TNBC 以及可能存在 TP53 缺失的许多其他类型的癌症非常有价值。