Engineering non-proliferating-but-active (NPBA) probiotic for breast-cancer gene therapy

用于乳腺癌基因治疗的工程非增殖但有活性（NPBA）益生菌

基本信息

批准号：
10533816
负责人：
RUSSELL C HOVEY
金额：
$ 16.73万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-12-02 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10533816
关键词：
Address Antibiotics Attenuated Autolysis BRCA1 gene Bacteria Breast Breast Cancer Cell Breast Cancer Model Breast Cancer Treatment Breast Cancer cell line Breast Cancer gene Breast Cancer therapy CD47 gene Cell Line Cell membrane Cells Clustered Regularly Interspaced Short Palindromic Repeats Cytosol DNA DNA delivery Data Development Disease Drug Delivery Systems Duct (organ) structure ERBB2 gene Effectiveness Engineering Environment Escherichia coli Essential Genes Estrogen Antagonists Face Foundations Future Gene Amplification Gene Delivery Gene Expression Genes Growth Hydrogels Infection Invaded Label Listeria monocytogenes Mammalian Cell Measures Membrane Metabolism Methods Molecular Molecular Target Monoclonal Antibodies Oncogenic Organ PIK3CA gene Pathway interactions Patients Penetration Pharmaceutical Preparations Phenotype Probiotics Proteins Proteome RNA Recombinants Research Resistance Site Solid Neoplasm Stress System Systemic Therapy T cell response T-Lymphocyte TP53 gene Testing Therapeutic Tissues Treatment Efficacy Work Yersinia pestis antagonist anti-cancer aqueous biomaterial compatibility cancer cell cancer immunotherapy cancer therapy crosslink cytotoxic delivery vehicle design drug resistance development effective therapy gene repair gene therapy genome editing in vivo information processing innovation loss of function mutation malignant breast neoplasm migration nanobodies novel strategies novel therapeutics public health relevance side effect therapeutic protein tool tumor

项目摘要

PROJECT SUMMARY Engineering non-proliferating-but-active (NPBA) probiotic for breast-cancer gene therapy A major breast-cancer treatment challenge is the debilitating effects of systemic therapies. To create more tolerable and effective therapies against breast cancer, considerable effort is being directed to using CRISPR- Cas to target the molecular origins of the disease. However, current methods face multiple challenges in delivering CRISPR-Cas to breast cancer cells, including low penetration of solid tumors, low delivery amount, and small size of CRISPR-Cas. To address the challenges, we will engineer a new kind of CRISPR-Cas delivery vehicle for breast cancer cells using non-proliferating-but-active (NPBA) probiotic. Our preliminary work demonstrates that the NPBA probiotic conduct metabolism, move in aqueous environments, and does not proliferate. We ultimately envisage that the NPBA probiotic can be introduced into the breast ducts or tumors to migrate towards cancer cells and then deliver a CRISPR-Cas construct that targets specific oncogenic mechanisms. Hence, this proposal aims to engineer these NPBA probiotic to deliver genes or proteins to breast cancer cells. Our team will leverage expertise in gene expression control, bacterial information processing, molecular tools, and breast cancer models. We will pursue two parallel aims that correspond to the engineering and testing of the NPBA Escherichia coli Nissle 1917. 1) Enable NPBA probiotics to invade mammalian cells before delivering CRISPR-Cas. This aim will establish the basic CRISPR-Cas-delivery function of the NPBA probiotics. 2) Enhance the invasion and CRISPR-Cas delivery efficacy of the NPBA probiotic in silencing the ErbB2 oncogene of breast cancer cells. This project will create a new paradigm that uses non-proliferating-but- active probiotics for effective and specific delivery of CRISPR-Cas to cancer cells. Our work will also form the foundation for using the NPBA probiotics to target various oncogenic and proliferative pathways of cancer cells. Future work may also investigate any dispersion of the NPBA probiotics into the surrounding tissues and throughout the host.

项目摘要乳腺癌基因疗法的工程非增殖 - 活性（NPBA）益生菌主要的乳腺癌治疗挑战是全身疗法的使人衰弱的作用。创建更多可忍受有效的针对乳腺癌的疗法，正在努力使用CRISPR- CAS靶向该疾病的分子起源。但是，当前的方法在向乳腺癌细胞提供CRISPR-CAS，包括实体瘤的渗透率低，递送量低，和少量CRISPR-CAS。为了应对挑战，我们将设计一种新型的CRISPR-CAS交付乳腺癌细胞的媒介物使用非增殖 - 活性（NPBA）益生菌的媒介物。我们的初步工作证明NPBA益生菌的代谢，在水性环境中移动，而不是增生。我们最终设想可以将NPBA益生菌引入乳房管道或肿瘤中向癌细胞迁移，然后提供针对特定致癌的CRISPR-CAS构建体机制。因此，该提案旨在设计这些NPBA益生菌以将基因或蛋白质传递给乳房癌细胞。我们的团队将利用基因表达控制，细菌信息处理的专业知识，分子工具和乳腺癌模型。我们将追求与工程相对应的两个平行目标和NPBA大肠杆菌Nissle 1917的测试。1）使NPBA益生菌能够入侵哺乳动物细胞在交付crispr-cas之前。这个目标将确定NPBA的基本CRISPR-CAS递送功能益生菌。 2）增强NPBA益生菌在沉默中的入侵和CRISPR-CAS递送功效乳腺癌细胞的ERBB2癌基因。该项目将创建一个新的范式，该范式使用非强化但活性益生菌可有效且特异性地递送CRISPR-CAS向癌细胞。我们的工作也将形成使用NPBA益生菌靶向癌细胞的各种致癌和增殖途径的基础。未来的工作还可以调查NPBA益生菌在周围组织中的任何分散在整个主机中。