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.

项目概要用于乳腺癌基因治疗的工程非增殖但有活性（NPBA）益生菌乳腺癌治疗的一个主要挑战是全身治疗的衰弱作用。去创造更多针对乳腺癌的可耐受且有效的治疗方法，人们正在努力使用 CRISPR- Cas针对疾病的分子起源。然而，当前的方法在以下方面面临多重挑战将CRISPR-Cas递送至乳腺癌细胞，包括实体瘤渗透率低、递送量低、以及小尺寸的 CRISPR-Cas。为了应对这些挑战，我们将设计一种新型 CRISPR-Cas 递送方法使用非增殖但有活性（NPBA）益生菌的乳腺癌细胞载体。我们的前期工作证明 NPBA 益生菌进行新陈代谢，在水环境中移动，并且不增生。我们最终设想可以将 NPBA 益生菌引入乳腺导管或肿瘤中，以向癌细胞迁移，然后递送针对特定致癌基因的 CRISPR-Cas 构建体机制。因此，该提案旨在改造这些 NPBA 益生菌，将基因或蛋白质输送到乳房癌细胞。我们的团队将利用基因表达控制、细菌信息处理、分子工具和乳腺癌模型。我们将追求与工程相对应的两个并行目标并测试 NPBA 大肠杆菌 Nissle 1917。 1) 使 NPBA 益生菌能够侵入哺乳动物细胞在交付 CRISPR-Cas 之前。该目标将建立 NPBA 的基本 CRISPR-Cas 传递功能益生菌。 2) 增强 NPBA 益生菌的侵袭和 CRISPR-Cas 传递功效，从而沉默乳腺癌细胞的 ErbB2 癌基因。该项目将创建一个新的范式，使用非扩散但- 活性益生菌可将 CRISPR-Cas 有效且特异性地递送至癌细胞。我们的工作也将形成为使用 NPBA 益生菌靶向癌细胞的各种致癌和增殖途径奠定了基础。未来的工作还可能调查 NPBA 益生菌在周围组织中的分散情况以及整个主机。