Excellence in Research: Elucidating the mechanisms that regulate cell uptake of homogeneous biodegradable polymeric nanoparticles to improve targeted therapeutic delivery

在过去的二十年里，针对多种疾病、感染和疫苗的治疗方法有了显著的增加。然而，在向特定组织和细胞提供治疗以及高剂量导致不必要的副作用方面，仍面临许多挑战。可生物降解的聚合物纳米粒为治疗药物和疫苗的输送提供了许多优点，如细胞和组织靶向、缓释和减少副作用以提高疗效。该项目的目标是证明，通过使用大小和形状相似的纳米颗粒，更好的细胞摄取和输送将导致更高的疗效。该项目的创新性有可能开发出安全和新颖的疗法，对由细菌、病毒、真菌和癌症引起的许多传染病有效。该项目将为阿拉巴马州立大学STEM领域中代表性不足的本科生和研究生提供研究培训和指导，这些学生来自纳米技术、免疫学、细胞生物学、基因组学和蛋白质组学等学科。学生还将获得其他教育机会，包括研讨会、专业发展培训活动，如研究中的道德规范、科学写作和在科学会议上的陈述。该项目将大大加强学生的研究和教育技能，为全球劳动力带来多样性。内吞作用是调节生物系统中可生物降解的聚合物纳米颗粒摄取和传输机制的主要途径之一，用于治疗输送。了解纳米颗粒的吞噬机制对于有效的治疗传递和功能至关重要，特别是关于它们的同质和异质物理性质。许多已知的治疗药物来自多相而不是均相的聚合物纳米颗粒。虽然这两种类型的纳米颗粒由于其物理性质可能具有不同的内吞机制，但关于调控均质纳米颗粒的细胞摄取以及对治疗递送和功能的影响的内吞机制的数据很少。需要检验的假设是，治疗递送效率取决于纳米颗粒的同质性；由于均一性和独特的吞噬机制，均质纳米颗粒比非均质纳米颗粒将更有效地被内吞，以改善治疗递送和功能。本项目将以细胞内蛋白为靶点，将衣原体细菌蛋白包裹在聚D、L-丙交酯-聚乙交酯和聚乳酸-聚乙二醇中，以评价其与树突状细胞作为抗原提呈细胞的相互作用。其目的是：1)制备和表征被包裹的蛋白质，以了解其与树突状细胞的相互作用，从而导致细胞功能的摄取和调节；2)破译不同大小和配方的被包裹的蛋白质所利用的特定的内吞途径，3)确定特定的内吞途径在小鼠体内调节被包裹的蛋白质的摄取和治疗功能的作用，以及4)将树突状细胞的发现扩展到其他细胞系。这项研究将利用各种机械方法来揭示新知识，从而获得改善人类健康相关疾病的科学发现。结果将显示内吞作用如何不同地调节均质和异质纳米粒的输送，以增强靶向输送和治疗功能。这一新知识将使科学家能够开发高效、均匀、大小特定的细胞内纳米载体，用于针对多用途生物医学应用的治疗。更广泛的影响是，这些发现将适用于更多类别的纳米颗粒，进一步推动新的科学知识，从而产生影响生物医学和生物制药应用的新型纳米疗法。该项目还将拓宽纳米技术的研究基础设施，并通过向阿拉巴马州未被充分代表的少数群体和整个社会提供优质教育和尖端技能来推进阿拉巴马州立大学的使命。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Over the past two decades, there has been a significant increase in therapeutics targeting multiple diseases, infections, and vaccines. However, many challenges are still being faced with therapeutic delivery to specific tissues and cells and high dosages causing unwanted side effects. Biodegradable polymeric nanoparticles offer many advantages for the delivery of therapeutic drugs and vaccines, such as cell and tissue targeting, slow-release, and reduced side effects to enhance the efficacy. The goal of this project is to show that by using nanoparticles of similar size and shape, better cell uptake and delivery will result in improved efficacy. The innovativeness of this project has the potential to develop safe and novel therapeutics that are effective for many infectious diseases caused by bacteria, viruses, fungi, as well as cancers. The project will provide research training and mentoring to underrepresented undergraduate and graduate students in STEM fields at Alabama State University from disciplines such as nanotechnology, immunology, cell biology, genomics, and proteomics. Students will also be provided other educational opportunities, including seminars, professional development training activities such as ethics in research, scientific writing, and presentations at scientific meetings. The project will considerably strengthen the research and educational skills of students to bring diversity to the global workforce.Endocytosis is one major route that regulates biodegradable polymeric nanoparticle uptake and transport mechanisms in biological systems for therapeutic delivery. Understanding the endocytosis mechanisms of nanoparticles is critical for effective therapeutic delivery and function, especially regarding their homogeneous and heterogeneous physical properties. Much of what is known for the delivery of therapeutics comes from heterogeneous rather than homogeneous polymeric nanoparticles. Although both nanoparticle types may have distinct endocytosis mechanisms due to their physical properties, there is a paucity of data on the endocytosis mechanism regulating homogeneous nanoparticles' cellular uptake and impact on therapeutic delivery and function. The hypothesis to be tested is that therapeutic delivery efficiency is dependent on the homogeneity of nanoparticles; homogeneous rather than heterogeneous nanoparticles will be more efficiently endocytosed to improve therapeutic delivery and function due to uniformity and distinct endocytosis mechanisms. This project will focus on intracellular protein targeting for delivery of a chlamydial bacterial protein encapsulated in poly D, L-lactide-co-glycolide, and poly(lactic acid)-poly(ethylene glycol to evaluate the interaction with dendritic cells as antigen-presenting cells in vitro and mice. The objectives are: 1) to formulate and characterize the encapsulated protein to understand its interactions with dendritic cells leading to uptake and regulation of cellular function; 2) to decipher the specific endocytosis pathway exploited by the encapsulated protein of different size and formulation, 3) to determine the role of specific endocytosis pathways regulating the encapsulated protein uptake and therapeutic function in mice, and 4) to extend the dendritic cells findings to other cell lineages. The research will exploit various mechanistic approaches to unveil new knowledge leading to scientific discoveries to improve human health-related diseases. The results will show how endocytosis differentially regulates the delivery of homogeneous and heterogeneous nanoparticles to enhance targeted delivery and therapeutic function. The new knowledge will enable scientists to develop efficient homogeneous size-specific intracellular nanocarriers for therapeutics targeting multipurpose biomedical applications. The broader impact is that the findings will apply to more categories of nanoparticles, further advancing new scientific knowledge leading to novel nanotherapeutics impacting biomedical and biopharmaceutical applications. This project will also broaden the research infrastructure in nanotechnology and advance the mission of Alabama State University by providing quality education and cutting-edge skills to the underrepresented minority population in Alabama and the society at large.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.