Deciphering the nanomaterial uptake in a bystander manner invoked by co-administration of functionalized nanoparticles

以旁观者的方式破译由功能化纳米颗粒共同施用引起的纳米材料吸收

• 批准号: 10170384
• 负责人: Hongbo Pang
• 金额: $ 30.42万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10170384
• 关键词: Amino Acids; Binding; Caliber; Cell Surface Receptors; Cell membrane; Cell physiology; Cell surface; Cells; Cellular biology; Complex; Coupled; Cysteine; Diagnosis; Drug Delivery Systems; Endocytosis; Environment; Gene Delivery; Genetic; Genetic Screening; Genetic Transcription; Goals; Heparan Sulfate Proteoglycan; Heparitin Sulfate; Ligands; Light; Link; Liquid substance; Mediating; Membrane; Metabolic; Metabolism; Methods; Molecular Profiling; Nanotechnology; Nutrient; Pathway interactions; Peptides; Play; Process; Property; RNA interference screen; Regulation; Research; Role; Signal Transduction; Site; Sulfate; Tissues; Vacuole; Validation; Vesicle; extracellular; genome-wide; human disease; in vivo; insight; nanomaterials; nanoparticle; nanoparticle delivery; nanoscale; novel; particle; receptor; synergism; uptake

Abstract One important yet understudied aspect in nanomaterial applications is the cell biology of nanomaterial transport into cells and through tissues. To overcome the cell membranes and reach their sites of action within, nanoparticles (NPs) are often complexed with cell-penetrating ligands. Such ligands include peptides like TAT (Trans-Activating Transcription). These ligands engage with cell surface receptors to invoke endocytic processes for NP uptake. However, the impact of these functionalized NPs and the resulting endocytic process on the cellular uptake of other NPs remains unknown. In this proposal, I aim to tackle this problem by studying TAT-coupled NPs (TAT-NP). By tracing NPs coated with several cell-penetrating peptides, including TAT, we previously discovered a novel receptor-dependent macropinocytosis (MP) pathway for NP uptake. MP has relatively large endocytic vacuoles (>200 nm in diameter) and thus can more readily engulf cargo as large as nanoparticles. Here, we found that TAT-NPs, by invoke this MP pathway, can bring into cells bystander NPs that are unable to enter the cells by themselves. This bystander uptake depends on TAT interaction with its receptor, heparan sulfate (HS) proteoglycans, and occurs through the HS-dependent MP pathway. We further showed that it is only active for NP-type bystander cargo, and its activity is greatly stimulated by Cysteine (Cys) outside the cells. Here, we aim to dissect out this bystander uptake process. In the Aim 1, we will determine the prerequisites for initiating the bystander uptake. In the Aim 2 and 3, we will utilize genetic screening and other methods to determine the factors that mediate this bystander uptake, and define its properties of cargo selectivity and Cys regulation. In the Aim 4, we will explore the potential use in biomedical applications. My proposed studies hold great potential for unveiling important cellular machineries for nanomaterial transport, boosting the efficiency of intracellular delivery, and opening up a new avenue to study cell biology, metabolism and nanomaterial delivery.

抽象的 纳米材料应用中的一个重要但细小的方面是纳米材料的细胞生物学 运输到细胞并通过组织。克服细胞膜并到达其作用位置 在内部，纳米颗粒（NP）通常与细胞渗透配体复合。这样的配体包括肽 像TAT（反式激活转录）。这些配体与细胞表面受体接合以调出 NP摄取的内吞过程。但是，这些功能化的NP和由此产生的影响 其他NP的细胞摄取上的内吞过程仍然未知。在这个建议中，我的目标是解决这个问题 通过研究TAT耦合的NP（TAT-NP）来解决问题。 通过追踪涂有几种细胞穿透肽（包括TAT）的NP，我们以前发现了A NP摄取的新型受体依赖性大型细胞增多症（MP）途径。 MP具有相对较大的内吞 液泡（直径> 200 nm），因此可以更容易地吞噬与纳米颗粒一样大的货物。在这里，我们 发现通过调用此MP途径的TAT-NP可以将无法进入的单元旁边的NP带入旁观者NP 单元自己。该旁观者的摄取取决于与其受体硫酸乙酰肝素相互作用 （HS）蛋白聚糖，并通过依赖HS的MP途径发生。我们进一步表明这只是 NP型旁观者货物的活性，其活性受到细胞外半胱氨酸（CYS）的极大刺激。 在这里，我们旨在剖析此旁观者的吸收过程。在目标1中，我们将确定先决条件 用于启动旁观者的吸收。在目标2和3中，我们将利用基因筛查和其他方法 确定介导该旁观者吸收的因素，并定义其货物选择性和 CYS调节。在目标4中，我们将探讨生物医学应用中的潜在用途。我提出的 研究具有揭示重要的纳米材料运输的重要细胞机械的巨大潜力 细胞内递送的效率，并为研究细胞生物学，代谢和 纳米材料输送。