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

• 批准号: 10350147
• 负责人: RUSSELL C HOVEY
• 金额: $ 20.66万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-02 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10350147
• 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; Drug resistance; 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; Heel; Hydrogels; Infection; Invaded; Label; Lead; 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 development; effective therapy; gene repair; gene therapy; genome editing; in vivo; information processing; innovation; loss of function mutation; malignant breast neoplasm; nanobodies; novel strategies; novel therapeutics; public health relevance; side effect; therapeutic protein; tool; tumor

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 用于乳腺癌基因治疗的工程化非增殖但活性（NPBA）益生菌 乳腺癌治疗的一个主要挑战是全身治疗的衰弱效应。创造更多 针对乳腺癌的可耐受和有效的疗法，相当大的努力是针对使用CRISPR- 以疾病的分子起源为目标。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，包括实体瘤的低穿透性，低递送量， 和小尺寸的CRISPR-Cas。为了应对这些挑战，我们将设计一种新的CRISPR-Cas交付方式， 使用非增殖但活性（NPBA）益生菌的乳腺癌细胞载体。我们的前期工作 证明NPBA益生菌进行代谢，在水性环境中移动，并且不 增殖。我们最终设想NPBA益生菌可以被引入乳腺导管或肿瘤中， 然后递送靶向特定致癌基因的CRISPR-Cas构建体， 机制等因此，该提案旨在工程化这些NPBA益生菌以将基因或蛋白质递送到乳房 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。我们将追求两个平行的目标， 和NPBA大肠杆菌Nissle 1917的测试。1)使NPBA益生菌能够侵入哺乳动物细胞 在交付CRISPR-Cas之前。这一目标将确立NPBA的基本CRISPR-Cas递送功能 益生菌2)增强NPBA益生菌在沉默细胞中的侵袭和CRISPR-Cas递送功效 乳腺癌细胞的ErbB 2癌基因。这个项目将创造一个新的范例，使用非扩散-但- 活性益生菌用于有效和特异性地将CRISPR-Cas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。 未来的工作还可能研究NPBA益生菌在周围组织中的任何分散， 整个主机。