CAREER: Branched Amphiphilic Peptide Capsules (BAPCs) for the delivery of lethal dsRNA into invasive organisms

迫切需要新的和创新的农药来解决一些挑战，包括对现有处理的抗性，对传粉媒介等重要物种的意外伤害以及环境破坏。一种很有希望的方法是使用双链RNA （dsRNA）来破坏昆虫的基本基因，特别是针对有害物种，同时保留有益物种。最近的研究表明，当dsRNA与纳米材料结合时，其有效性显着提高。这种组合防止了dsRNA的过早降解，并帮助它被昆虫肠道的细胞吸收。本项目旨在探索纳米材料的特定性质如何影响基于dsrna的生物农药的性能。研究人员将使用一种由多肽制成的独特纳米材料，研究大小、电荷和组成的变化如何影响dsRNA的吸收、昆虫的生存和在不同环境条件下的稳定性。这项研究的结果意义重大，因为它们促进了我们对设计农业用途生物材料的理解，有可能导致新的安全农药的产生。此外，该项目旨在通过实践体验让学生参与科学，特别是那些来自历史上被排斥群体的学生。研究小组将通过奥本大学的项目，教育阿拉巴马州的学生纳米材料作为杀虫剂的潜力。参与这些实验的高中生将提高他们对科学的理解和兴趣，有助于他们参与STEM领域。该项目由生物材料计划和促进竞争性研究的既定计划（EPSCoR）共同资助。技术概述采用替代策略进行虫害管理对于应对农药耐药性、脱靶效应和环境危害带来的挑战至关重要。利用双链RNA （dsRNA）敲除昆虫的必需基因已成为一种有前景的替代传统农药的方法。这种方法针对有害物种而不影响有益物种，缺乏已知的昆虫抗性机制。最近的发现表明，dsRNA与纳米材料的关联通过防止过早降解和促进肠道细胞的吸收来增强其功效。本研究计划的主要目的是探索纳米材料特性如何影响基于dsrna的生物农药的致死效应。具体来说，本项目涉及将支链两亲肽胶囊（BAPCs）与dsRNA结合使用。BAPCs是由PI开发的一类新型生物材料，在纳米颗粒递送系统的竞争中脱颖而出，因为两个重要因素：1)BAPCs只在水中组装，2)BAPCs含有4个游离赖氨酸Ɛ-amino基团，pKa值在9到10.5之间，这使得它们在中性和碱性昆虫肠道中稳定。先前的数据表明，BAPC-dsRNA复合物靶向Tribolium castaneum和Acyrthosiphon pisum的必需基因，导致这两种物种的高死亡率。该提案假设，通过操纵bapc的大小、电荷和组成，有可能调节：1)dsRNA的细胞摄取和分布，以及它在中肠细胞中的运输，2)昆虫的生存，以及3)对环境因素的稳定性。秋粘虫（Spodoptera frugiperda）和棉蚜（Aphis gossypii）将作为检验这一假设的害虫模型。除了科学探索之外，该提案还具有教育目的。它旨在通过K-12外展倡议教育阿拉巴马州的学生关于纳米材料在害虫控制方面的潜力。学生将积极参与与研究相关的活动，促进STEM领域的参与。此外，本提案旨在促进对农业用途生物材料设计的基本理解，并弥合关于胞吞作用在纳米材料在昆虫内脏运输中的作用的知识空白。该项目由生物材料计划和促进竞争性研究的既定计划（EPSCoR）共同资助。该奖项反映了美国国家科学基金会的法定使命，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。

NON-TECHNICAL SUMMARYNew and innovative pesticides are urgently needed to address several challenges including resistance to current treatments, unintended harm to important species like pollinators, and environmental damage. One promising approach involves using double-stranded RNA (dsRNA) to disrupt essential genes in insects specifically targeting harmful species while sparing beneficial ones. Recent studies have revealed that when dsRNA is combined with nanomaterials, its effectiveness significantly increases. This combination prevents premature degradation of dsRNA and helps it get absorbed into the cells of insect guts. This project aims to explore how specific properties of nanomaterials influence the performance of dsRNA-based biopesticides. Researchers will work with a unique nanomaterial made of peptides and study how variations in size, charge, and composition impact dsRNA absorption, insect survival, and stability in different environmental conditions. The outcomes of this study are significant as they advance our understanding of designing biomaterials for agricultural use, potentially leading to the creation of new safe pesticides. Additionally, the project aims to involve students in science, especially those from historically excluded groups through hands-on experiences. The research team will educate students in Alabama about the potential of nanomaterials as pesticides through programs at Auburn University. High school students participating in these experiments will improve their understanding and interest in science contributing to their engagement in STEM fields.This project is jointly funded by the Biomaterials Program and the Established Program to Stimulate Competitive Research (EPSCoR).TECHNICAL SUMMARYAdopting alternative strategies for insect pest management is critical to counter challenges posed by pesticide resistance, off-target effects, and environmental harm. Utilizing double-stranded RNA (dsRNA) to knockdown essential genes in insects has emerged as a promising alternative to conventional pesticides. This method targets harmful species without affecting beneficial ones and lacks known insect resistance mechanisms. Recent discoveries demonstrated that association of dsRNA with nanomaterials enhances its efficacy by preventing premature degradation and facilitating uptake by gut cells. The primary objective of this research proposal is to explore how nanomaterial properties influence the lethal effects of dsRNA-based biopesticides. Specifically, this project involves using Branched Amphiphilic Peptide Capsules (BAPCs) combined with dsRNA. BAPCs are a new class of biomaterial developed by the PI that stand out in the crowded field of nanoparticle delivery systems owing to two important factors: 1) BAPCs are assembled exclusively in water, and 2) BAPCs contain four free lysine Ɛ-amino groups with pKa values between 9 and 10.5, which makes them stable in neutral and alkaline insect guts. Previous data demonstrated that BAPC-dsRNA complexes target essential genes in Tribolium castaneum and Acyrthosiphon pisum, leading to high mortality rates in both species. The proposal hypothesizes that by manipulating the size, charge, and composition of BAPCs, it's possible to regulate: 1) the cellular uptake and distribution of dsRNA, as well as its transport across midgut cells, 2) insect survival, and 3) stability against environmental elements. The fall armyworm (Spodoptera frugiperda) and the cotton aphid (Aphis gossypii) will serve as pest models for testing the hypothesis. Beyond the scientific exploration, the proposal also has educational objectives. It seeks to educate Alabama students about the potential of nanomaterials in pest control through K-12 outreach initiatives. Students will actively participate in research-related activities, promoting engagement in STEM fields. Additionally, this proposal aims to contribute to the fundamental understanding of biomaterial design for agricultural purposes and bridge knowledge gaps regarding the role of transcytosis in nanomaterial transport across insect midguts.This project is jointly funded by the Biomaterials Program and the Established Program to Stimulate Competitive Research (EPSCoR).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.