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

PI：Behkam，巴哈伊提案编号：1454226癌症是美国第二大死亡原因，每年夺去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使毒副作用最小化的更有效的药物递送技术。工程细菌已被证明可以安全地在肿瘤组织中以高选择性积累，并治疗对小鼠癌症模型中的常规疗法无反应的癌症。然而，由于细菌的肿瘤定植不足，临床成功是罕见的。目前，这归因于宿主免疫应答，特别是细菌-嗜中性粒细胞相互作用已在几项体内研究中涉及。尽管如此，肿瘤微环境中嗜中性粒细胞和细菌的相互作用在很大程度上仍未被探索。该CAREER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，目标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学障碍。拟议的CAREER研究和教育计划有可能通过实现新的癌症治疗途径来彻底改变生活质量，同时提供将研究元素和发现整合到多学科教育和推广经验中的机会。该CAREER提案的研究部分的总体目标是生物制造一种基于活的、用负载白细胞毒素的纳米颗粒减弱肿瘤靶向细菌，以便能够局部控制肿瘤微环境内的细菌-免疫细胞相互作用。据推测，通过局部调节嗜酸性粒细胞-细菌相互作用来控制肿瘤内炎症将使细菌能够克服免疫屏障，从而在活的肿瘤组织中增殖。将对工程3D疾病模型中的细胞-细胞相互作用进行基础研究，并开发控制这种相互作用的方法。 本论文的主要研究目标如下：（1）纳米细菌免疫调节系统的生物制造和表征（2）研究细菌生物膜形成和中性粒细胞刺激的相互作用在细菌肿瘤内定植中的作用;和（3）定量由NanoBEIMS使能的嗜中性粒细胞的局部消耗引起的肿瘤中细菌增殖的增强。通过这种跨学科的努力获得的新知识将提供一个独特的使能技术库，用于局部细胞-细胞相互作用调节，以及癌症治疗及其他领域的高度靶向治疗诊断应用。免疫系统-细菌相互作用的知识对于阐明细菌肿瘤发生的基础和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。该职业计划教育部分的主要目标是加强STEM中社会经济弱势群体和种族代表性不足群体的招聘和保留，并为未来的科学家应对生物/非生物系统工程（BASE）中复杂的多学科挑战做好准备。为此，本CAREER提案将建立一个以研究型学习为中心的多层计划。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）为社区学院学生提供多学科研究经验，特别是针对代表性不足的本科生。

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.