Genetically ENgineered BIOsensors to detect BIological Threats (GENBIOBIT): Influenza A Virus

我们希望开创一种新型的基因工程生物传感器，能够快速、灵敏和高效地检测和诊断传染病，以应对快速变化的人类和动物种群生物监控格局。传染病和新出现的生物威胁(自然的、意外的或故意的)是对英国安全的挑战。我们的目标是实现当前技术能力的阶段性变化，以克服目前阻碍基于细胞的生物传感器在不进行常温下长时间样品制备的情况下在现场工作的障碍。这项新技术可能成为具有重大经济和社会影响的多种生物威胁的全国性生物监测网络的一部分。特别是，我们希望建立甲型流感病毒可行性的证据。IAV已被证明具有破坏牲畜福利和生产力以及引起人类大流行的潜力。它们在野生动物种群中茁壮成长，并可以传播给养殖动物。除了造成直接的经济损害外，这还为人畜共患病(例如禽流感H5N1亚型和猪H1N1亚型)提供了通道。有记录以来最严重的一次是1918年的大流行，导致5000多万人死亡。为了减少经济影响和公共卫生风险，有必要监测和控制动物来源的疾病。尽管实验室已经开发了多种IAV生物传感器，但目前还没有实际应用。我们希望引入一项关键创新，使它们能够在该领域使用，特别是在动物饮用者中。关键的创新是一种合成膜受体，1)具有对IAVS具有高度选择性的胞外结构域，2)具有细胞内结构域，在病毒结合时激活控制细胞行为的特定感兴趣基因3)可以在源自虹鱼鳃的弹性细胞系中表达，该细胞系被证明可以在一年多的温度范围内存活，从野外无需维护到测试水毒性。最后，通过组合多电极阵列和多个工程细胞系(对病毒具有相同的选择性，但编码不同的细胞行为)，我们将拥有一个多路电子读数，大大提高其稳定性和区分(诊断)IAV与其他生物威胁(毒素、细菌、病毒)的潜力，比与病原体相关的细胞死亡相关的信号具有更快的反应时间和灵敏度。手机大小的混合传感器(包含由多电极阵列并行询问的几个基因工程细胞)将在实验室(TRL3)中演示，以检测和区分IAV和其他病原体或毒素。我们的方法利用合成生物学和数据科学来监测现有的传染病，从而迅速适应新出现的生物威胁。合成受体的胞外部分可以迅速重新设计，以应对其他生物威胁。基因工程生物传感器将为生物传感器网络铺平道路，这些生物传感器网络可以部署在现场对水(饮水器、池塘、湖泊)或挥发物(例如在空调系统中)进行采样，并可以直接向监测中心发出警报或提供实时数据。这项通过便携式生物传感器检测IAV的新技术也将对生命科学界产生更广泛的影响，因为生命科学界非常需要检测靶标结合的新工具(例如药物筛选、传染病)。这也将为新兴的生物计算领域提供一个新的工具。这是一个高度跨学科的项目，对所涉及的PDRA来说，具有跨越病毒学、合成生物学和生物传感学科的巨大潜力。

We want to pioneer a new class of genetically engineered biosensors that can rapidly, sensitively and efficiently detect and diagnose infectious diseases to address the fast-changing landscape of human and animal population biosurveillance. Infectious diseases and emerging biological threats (natural, accidental or deliberate) are a challenge to UK security. We aim to deliver a step-change in current technological capability to overcome the current barriers that prevent cell-based biosensors to work in the field without sample preparation at ambient temperature for a prolonged period of time. This new technology could be part of a nationwide biosurveillance network of multiple biological threats with large economic and societal impact. In particular, we want to establish proof-of-feasibility for the influenza A virus. IAVs have proven potential to damage livestock welfare and productivity, as well as to cause human pandemics. They thrive in wild animal populations and can transmit to farmed animals. As well as causing direct economic harm, this then provides a gateway for zoonotic infections (e.g. avian H5N1 and swine H1N1 subtypes). The worst recorded instance is the 1918 pandemic that resulted in the deaths of more than 50 million people. To reduce economic impacts as well as public health risk, there is a need to monitor and control the disease in the animal source. Despite multiple IAVs biosensors have been developed in the lab, there is currently no practical applications. We wish to introduce a key innovation that will enable their use in the field, and in particular in animal drinkers. The key innovation is a synthetic membrane receptor that 1) has an extracellular domain that is highly selective to IAVs 2) has an intra-cellular domain that activates specific genes of interest controlling cell behavior upon virus binding 3) can be expressed in a resilient cell line originating from rainbow trout gills, that was demonstrated to survive over a range of temperature from more than a year in the field without maintenance to test water toxicity. Finally, by combining multielectrode arrays and multiple engineered cell lines (with the same selectivity to the virus but encoding different cell behavior) we will have a multiplexed electronic readout improving considerably the robustness and its potential to discriminate (diagnose) IAVs from others biological threats (toxins, bacteria, viruses) with faster response time and sensitivity than signals associated with pathogen associated cell death. Phone-sized hybrid sensors (containing several gene-engineered cells interrogated in parallel by multi-electrode arrays) will be demonstrated in the lab (TRL3) to detect and discriminate between IAVs and other pathogens or toxins. Our approach harnesses synthetic biology and data science to monitor existing infectious diseases and thus rapidly adapt to emerging biological threats. The extra-cellular part of the synthetic receptor can be quickly re-engineered to address other biological threats. Genetically engineered Biosensors will pave the way towards networks of biosensors that can be deployed in the field sampling water (drinkers, ponds, lakes) or volatiles (e.g. in air conditioning system) and could directly alert or feed real-time data to monitoring centres. This novel technology, demonstrated with a portable biosensor detecting IAV, will also provide a broader impact on the life science community for which novel tools for the detection of target binding are highly desirable (e.g. drug screening, infectious diseases). This will also provide a new tool for the emerging field of bio-computation. This is a highly interdisciplinary project with great potential for the PDRAs involved to work across the discipline of virology, synthetic biology and biosensing.