CAREER: Systems Microbiology and InterdiscipLinary Education for Halting Environmental Antibiotic Resistance Transmission (SMILE HEART)

细菌抗生素耐药性（AR）感染是全球公共卫生的十大威胁之一。仅在美国，每年报告的细菌性AR感染就超过280万例，造成至少35，000人死亡和约550亿美元的经济损失。细菌AR具有环境起源，在生态系统内和生态系统之间发展和传播，并通过环境暴露和/或食物和饮用水的摄入对人类健康构成风险。最近的研究已经确定，水平基因转移（HGT）介导不通过遗传连接的个体细胞之间的DNA交换，在控制环境中和临床相关的细菌病原体中抗生素耐药基因（ARG）的发展和传播中是关键的。该CAREER项目的总体目标是设计，开发和评估新的解决方案，以减轻和抑制AR基因在农业土壤中的传播及其通过HGT向病原体的潜在转移。为了推进这一目标，主要研究者提出测试以下假设：1）环境相关浓度的非抗生素微污染物（NAMP）混合物促进农业土壤中ARG通过HGT的传播，2）这种不良结果可以通过抑制IV型分泌系统（T4 SS）来减轻，T4 SS是原核生物中介导HGT的生物组装，使用天然产物罗特勒林。该项目的成功完成将通过产生新的基础知识来促进设计和部署更具成本效益和可持续性的解决方案，以减轻和防止农业土壤中ARG的发展和传播，从而使社会受益。通过学生教育和培训，包括指导州立大学纽约环境科学与林业学院的一名研究生和一名本科生，将为社会带来额外的好处。传播，是与抗生素耐药基因（ARG）的环境水库的风险相关的挑战的核心。在炎热的时刻，如农业土壤接受粪肥或生物固体热点HGT升级代表着一个重大的风险。 虽然广泛的非抗生素微污染物（NAMP），现在已被检测到在原料和处理过的粪肥生物固体和土壤接收粪肥或生物固体，鲜为人知的是，这些微污染物对HGT介导的ARGs在农业土壤中的传输的影响。这个职业生涯项目将解决这些关键的知识差距。本研究的具体目标是：1）从机理上理解NAMP在模拟农业土壤中诱导的HGT（2）通过结合多组学分析、微流体辅助原位观察和数据科学，确定农业土壤中NAMP诱导的HGT范围的生态进化控制;（3）证明rottlerin（一种多酚天然产物）作为农业土壤中HGT的广谱抑制剂的有效性;（4）阐明rottlerin在农业土壤和复杂微生物系统中HGT缓解作用的分子-群落机制，重点是网络相互作用。该项目的成功完成有可能通过产生新的基础知识产生变革性影响，以推动设计和实施具有成本效益的解决方案，以控制和减轻NAMP引起的农业土壤中的汞的汞浓度变化。为了实现这个职业生涯项目的教育和培训目标，主要研究者（PI）建议利用纽约州立大学环境科学与林业学院（ESF）正在进行的项目，以及与纽约北部的多个合作伙伴的合作，以（1）开发跨学科课程，实施团队科学，以促进ESF学生之间的跨学科合作，以及（2）设计并提供垂直整合的暑期课程（微笑振作），以扩大对STEM的参与。 此外，PI计划利用该项目的研究成果来设计高中课程模块，并为教师组织一次研讨会，以促进教育模块在纽约州的传播。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Bacterial antibiotic resistance (AR) infections are among the top 10 threats to global public health. In the United States alone, more than 2.8 million bacterial AR infections are reported each year, causing at least 35,000 deaths and approximately $55 billion in economic losses. Bacterial AR has an environmental origin, develops, and transmits within and between ecosystems and poses a risk to human health through environmental exposure and/or ingestion of food and drinking water. Recent studies have established that horizontal gene transfer (HGT) that mediates DNA exchange between individual cells not connected by inheritance is pivotal in controlling the development and transmission of antibiotic resistance genes (ARGs) in the environment and to bacterial pathogens of clinical relevance. The overarching goal of this CAREER project is to design, develop and evaluate new solutions to mitigate and curb the transmission of AR genes in agricultural soils and their potential transfer to pathogens via HGT. To advance this goal, the Principal Investigator proposes to test the hypothesis that 1) mixtures of non-antibiotic micropollutants (NAMPs) at environmentally relevant concentrations promote the transmission of ARGs in agricultural soils via HGT and 2) this adverse outcome can be mitigated by inhibiting the type IV secretion system (T4SS), the biological assembly that mediates HGT in prokaryotes, using the natural product rottlerin. The successful completion of this project will benefit society through the generation of new fundamental knowledge to advance the design and deployment of more cost-effective and sustainable solutions to mitigate and prevent the development and transmission of ARGs in agricultural soils. Additional benefits to society will be achieved through student education and training including the mentoring of one graduate student and one undergraduate student at the State University of New York College of Environmental Science and Forestry.Horizontal gene transfer (HGT) plays an essential role in antibiotic resistance (AR) dissemination and is at the heart of the challenge associated with the risk of environmental reservoirs of antibiotic resistance genes (ARGs). Escalated HGT in hotspots during hot moments such as agricultural soils receiving manure or biosolids represents a significant risk. Although a broad range of non-antibiotic micropollutants (NAMPs) have now been detected in raw and treated manure biosolids and soils receiving manure or biosolids, little is known about the impact of these micropollutants on HGT-mediated transmission of ARGs in agricultural soils. This CAREER project will address these critical knowledge gaps. The specific objectives of the research are to 1) develop a mechanistic understanding of NAMP-induced HGT in model agricultural soils (alfisols and mollisols) by combining model-assisted quantification and transcriptome analysis; (2) identify eco-evolutionary controls of the scope of NAMP-induced HGT in the model agricultural soils by combining multi-omics analyses, microfluidic-assisted in situ observations and data science, and; (3) demonstrate the effectiveness of rottlerin (a polyphenol natural product) as a broad-spectrum inhibitor of HGT in agricultural soils; and (4) elucidate the molecular-to-community mechanisms of the HGT mitigating effect of rottlerin in agricultural soils and complex microbial systems with a focus on network interactions. The successful completion of this project has the potential for transformative impact through the generation of new fundamental knowledge to advance the design and implementation of cost-effective solutions to control and mitigate NAMP-induced HGT of ARGs in agriculture soils. To implement the educational and training goals of this CAREER project, the Principal Investigator (PI) proposes to leverage ongoing programs at the State University of New York College of Environmental Science and Forestry (ESF) and collaborations with multiple partners in Upstate New York to (1) develop an interdisciplinary curriculum that implements team science to foster cross-disciplinary collaboration among ESF students and 2) design and deliver a vertically integrated summer program (SMILE HEART UP) to broaden participation in STEM. In addition, the PI plans to leverage the research findings from this project to design high school course modules and organize a workshop for teachers to facilitate the dissemination of the education modules across the State of New York.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.