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.

项目摘要/摘要乳腺癌是美国妇女与癌症相关死亡的第二大原因。特别是临床上几乎所有三重阴性乳腺癌（TNBC）都可以使用靶向疗法。癌症作为一个积累遗传改变的结果。因此，制定新型策略以精确针对遗传 TNBC的改变对于对抗恶性疾病可能是有价值的。 TP53是一个关键的肿瘤抑制剂大多数人类癌症中突变或缺失灭活的基因。已经付出了巨大的努力来恢复 TP53编码的p53蛋白的活性用于癌症治疗。不幸的是，没有基于p53的疗法由于p53信号的复杂性，成功地转化为诊所。因此，识别 TP53删除而不是恢复p53活动赋予的漏洞是打击的新策略癌症。在我们最近发表的自然和自然纳米技术的工作中，我们揭示了基因组缺失 TP53通常伴随着邻近基因Polr2a的半合子（即部分）损失对细胞必不可少的生存，几乎所有53％TNBC具有TP53缺失的Harbour Harbous Hemizygous Polr2a损失（TP53/polr2aloss）。我们的初步数据表明，通过RNA干扰小型干扰，抑制POLR2A的表达 RNA（siRNA）使用低pH激活的纳米物组选择性地抑制增殖，存活和 TP53/polr2aloss TNBC细胞的致瘤潜力。纳米炸弹在血液中保护siRNA并启用 siRNA进入siRNA执行其polr2a抑制函数的siRNA的内膜/溶酶体逃逸细胞吸收后。此外，纳米栓介导的靶向polr2a靶向siRNA的递送有选择地抑制原位TP53/polr2aloss TNBC肿瘤的生长，没有明显的全身毒性。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与变体CD44过表达（注意：不是非 - 已证明在正常干细胞上的变异或正常CD44对临床使用特别抗性 TNBC的化学疗法药物，例如紫杉醇（PTX）。由于polr2a对于癌细胞来说是必不可少的我们假设靶向POLR2A靶向siRNA和PTX的靶向共传递到变体 CD44+癌细胞可以克服TNBC耐药性。我们将进一步发展上述低没有主动靶向的pH激活的纳米泡，能够主动靶向变体CD44+ 细胞和肿瘤脉管系统。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，我们将使用使用POLR2A靶向治疗的抗性机制不仅是2D和异种移植，还使用美国开发的微流体方法生成的3D TNBC模型。总的来说，该项目可能会导致一种新型的耐药性TNBC疗法，并具有机械理解，这对于打击TNBC以及可能具有TP53缺失的许多其他类型的癌症是无价的。