CAREER: A Biomanufactured Platform for Modulating Immune Cell-Bacteria Interactions in the Tumor Microenvironment

PI：Bahkam，BaharehProposal编号：1454226癌症是美国第二大死因，每年夺走50多万人的生命。因此，迫切需要改进的非侵入性疾病诊断和更有效的药物输送技术，将毒副作用降至最低。在小鼠癌症模型中，工程菌已被证明可以安全地高选择性地在肿瘤组织中积聚，并治疗对传统疗法无效的癌症。然而，由于细菌对肿瘤的定植不足，临床上的成功一直很少。目前这被归因于宿主免疫反应，特别是细菌-中性粒细胞的相互作用在几个体内研究中被涉及。然而，中性粒细胞和细菌在肿瘤微环境中的相互作用在很大程度上仍未被探索。该职业奖专注于三维(3D)疾病模型中细菌-中性粒细胞相互作用的基础研究，目的是调节这些相互作用，以克服成功的基于细菌的癌症治疗的免疫障碍。拟议的职业研究和教育计划有可能通过启用新的癌症治疗途径来革命性地提高生活质量，同时提供机会将研究元素和发现整合到多学科的教育和外展体验中。这项职业计划研究部分的总体目标是生物制造一个基于活的、减毒的肿瘤靶向细菌与装载白毒素的纳米颗粒的受控自组装系统，以便能够在肿瘤微环境中局部控制细菌与免疫细胞的相互作用。据推测，通过局部调节中性粒细胞与细菌的相互作用来控制肿瘤内的炎症，将使细菌能够克服活肿瘤组织中增殖的免疫障碍。将对工程化3D疾病模型中的细胞-细胞相互作用进行基础研究，并将开发控制这种相互作用的方法。将致力于实现以下研究目标：(1)制备并表征首创的纳米级细菌免疫调节系统(NanoBEIMS)，用于局部和原位消除细菌在肿瘤内增殖的免疫屏障；(2)研究细菌生物膜形成和中性粒细胞刺激在肿瘤内定植中的相互作用；(3)量化由NanoBEIMS启用的中性粒细胞局部耗尽导致的肿瘤中细菌增殖的增强。通过这一跨学科努力获得的新知识将提供独特的使能技术，用于本地化的细胞-细胞相互作用调制，以及用于癌症治疗和其他领域的高度针对性的治疗应用。了解免疫系统与细菌的相互作用对于阐明细菌致癌的基础以及实现有效的癌症免疫治疗以及细菌介导的基因治疗也至关重要。这一职业规划的教育部分的主要目标是促进在STEM招聘和留住社会经济上处于不利地位和族裔代表性不足的群体，并使未来的科学家做好准备，以应对生物/非生物系统工程(BASE)中复杂的多学科挑战。为此，这份职业计划将建立一个以研究性学习为中心的多层次计划。该计划的三个主要组成部分包括：(1)为弗吉尼亚州西南部一个农村县的小学生和教师提供研究和培训机会；(2)基于网络的互动学习模块，向9-12岁的儿童介绍国家一级的非传统工程领域；(3)为社区大学学生提供的多学科研究经验，特别关注未被充分代表的本科生。

PI: Behkam, BaharehProposal Number: 1454226Cancer is the second leading cause of death in the U.S., annually claiming over half a million lives. Thus, there is an acute need for improved non-invasive disease diagnostics and more effective drug delivery techniques that minimize toxic side effects. Engineered bacteria have been shown to safely accumulate in tumor tissue with high selectivity and treat cancers that are not responsive to conventional therapies in mouse cancer models. However, clinical success has been rare due to insufficient tumor colonization by bacteria. This is presently attributed to the host immune response, and bacteria-neutrophil interactions in particular have been implicated in several in vivo studies. Nonetheless, interactions of neutrophil and bacteria within the tumor microenvironment remain largely unexplored. This CAREER award focuses on fundamental research in bacteria-neutrophil interactions in a three-dimensional (3D) disease model with the goal of modulating these interactions in order to overcome the immunologic barrier to successful bacteria-based cancer therapy. The proposed CAREER research and education plan has the potential to revolutionize the quality of life by enabling novel cancer treatment pathways, while providing the opportunity to integrate research elements and discoveries into multidisciplinary educational and outreach experiences.The overarching goal of the research component of this CAREER proposal is to biomanufacture a system based on controlled self-assembly of live, attenuated tumor-targeting bacteria with leukotoxin-loaded nanoparticles in order to enable localized control of bacteria-immune cell interactions within the tumor microenvironment. It is hypothesized that controlling intratumoral inflammation through localized modulation of neutrophil-bacteria interactions will enable bacteria to overcome the immunologic barrier to proliferation in viable tumor tissue. Fundamental studies on cell-cell interactions in engineered 3D disease models will be performed and means for controlling such interactions will be developed. The following research objectives will be pursued: (1) Biomanufacture and characterize a first-of-its-kind Nanoscale Bacteria Enabled Immune Modulation System (NanoBEIMS) for localized and in-situ disabling of the immunologic barrier to intratumoral proliferation of bacteria; (2) Investigate the role of the mutual effects of bacteria biofilm formation and neutrophil stimulation in intratumoral colonization of bacteria; and (3) Quantify enhancement in bacterial proliferation in tumors caused by NanoBEIMS-enabled localized depletion of neutrophils. The new knowledge gained through this interdisciplinary effort will provide a unique repertoire of enabling technologies for localized cell-cell interaction modulation, as well as highly-targeted theranostic applications for cancer therapy and beyond. Knowledge of the immune system-bacteria interactions is also critical to elucidating the underpinnings of bacterial oncogenesis and enabling effective cancer immunotherapy as well as bacteria-mediated gene therapy. The main goals of the educational component of this CAREER plan are to enhance recruitment and retention of the socioeconomically disadvantaged and ethnically underrepresented groups in STEM and to prepare future scientists to tackle complex multidisciplinary challenges in biotic/abiotic systems engineering (BASE). To this end, this CAREER proposal will establish a multi-tier plan centered on research-inspired learning. The three major components of the plan include: (1) research and training opportunities for elementary school students and teachers from a rural county in southwest Virginia, (2) web-based interactive learning modules to introduce 9-12 year old children to non-traditional areas in engineering at the national level; (3) multidisciplinary research experience for community college students with special focus on underrepresented undergraduates.