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

项目总结 用于乳腺癌基因治疗的非增殖活性益生菌(NPBA)工程 乳腺癌治疗的一个主要挑战是全身治疗的衰弱效果。要创造更多 针对乳腺癌的可耐受和有效的治疗方法，正在做出相当大的努力来使用CRISPR- CA以疾病的分子起源为目标。然而，当前的方法面临着多重挑战 向乳腺癌细胞递送CRISPR-CA，包括实体瘤的低渗透率，低释放量， 和小尺寸的CRISPR-CA。为了应对挑战，我们将设计一种新的CRISPR-CAS交付 使用非增殖但活性(NPBA)益生菌的乳腺癌细胞的载体。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，而不是 扩散。我们最终设想，NPBA益生菌可以被引入乳房导管或肿瘤中 向癌细胞迁移，然后递送靶向特定致癌基因的CRISPR-CAS构建体 机械装置。因此，这项提议旨在改造这些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 和NPBA大肠杆菌Nissle 1917的检测。1)使NPBA益生菌能够入侵哺乳动物细胞 在交付CRISPR-CA之前。这一目标将建立NPBA的基本CRISPR-CAS-交付职能 益生菌。2)增强NPBA益生菌的侵袭和CRISPR-Cas传递效率，使其沉默 乳腺癌细胞的ERBB2癌基因。这个项目将创造一种新的范式，使用非扩散-但- 有效和特异地将CRISPR-CA递送到癌细胞的活性益生菌。我们的工作也将形成 为使用NPBA益生菌靶向肿瘤细胞的各种致癌和增殖途径奠定基础。 未来的工作还可能调查NPBA益生菌在周围组织中的任何分散情况 在整个宿主中。

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.