Understanding and controlling the cellular fate of fluorine-modified biologics

了解和控制氟改性生物制品的细胞命运

基本信息

批准号：
10651637
负责人：
Scott Hammond Medina
金额：
$ 38.17万
依托单位：
PENNSYLVANIA STATE UNIVERSITY, THE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2026-06-30
项目状态：
未结题

项目摘要

PROJECT SUMMARY Organofluorine compounds possess attractive chemical, pharmacological and biological properties that have allowed them to make paradigm shifts in the design of biopharmaceuticals and biologic materials. The introduction of fluorine atoms into amino acids and nucleic acids opens a vast new chemical landscape with which to alter the folding, stability, oligomerization propensity and bioactivity of peptides, proteins and DNA. However, although shown to impart favorable properties, the impact of adding fluorine into biologic scaffolds is rarely predictable. Further, increasing evidence suggests perfluorinated compounds promiscuously adsorb to many of the fundamental building blocks of cells - including lipids, proteins and DNA - to elicit a plurality of bioeffects. One of these effects, recently discovered by the PI’s lab, is the ability of organofluorine molecules to direct protein and DNA assembly into fluorous microdomains that phase separate into fluorinated liquids without denaturing the biologic. The PI has recently exploited these emergent properties to enable ultrasound-guidance of fluorinated proteins in three-dimensional tissues. Building upon these preliminary findings, the proposed research program will mechanistically explore how organofluorine compounds influence the structure and function of adsorbed proteins and DNA and use these insights to guide the design of new supramolecular assembled biomaterials. Our overarching hypothesis is that organofluorine compounds non-covalently adsorb to proteins and DNA to direct their separation into fluorine-rich phases, which in turn alters their oligomeric assembly, cellular fate and bioactivity. To test this assertion, we will expand our perfluorinated compound (PFC) library to include a diversity of molecules with basic/acidic functionalities and heterocyclic moieties. We will use this library to establish structure-activity relationships governing the ability of PFCs to adsorb proteins, and investigate how PFC complexation alters protein cellular uptake, intracellular trafficking and bioactivity. In parallel, we will use this library to study the molecular mechanisms mediating PFC-DNA interactions and examine how PFC complexation alters DNA stability and metabolic homeostasis in exposed cells. Together, these studies will establish a comprehensive mechanistic understanding of how PFCs interact with proteins and DNA and will allow us to rationally design fluorous biotechnologies that exploit the unusual assembly phenomenon and phase- separation properties that emerge. As an example, we will create ultrasound-sensitive fluorine nanoemulsions loaded with PFC-modified ribonucleoproteins (RNPs) to enable imaging-guided gene editing in kidney tissue. Success of this research will advance the use of PFCs as a new molecular motif to control protein and DNA assembly, and the methods developed applied to discover new reagents for intracellular transduction of fluorinated biomacromolecules. Ultimately, advancing knowledge on how organofluorine compounds interact with proteins and DNA, and its effects on cells, will guide the rational design of new PFC enabled technologies with desirable functional properties for drug discovery and nanomedicine applications.

项目概要有机氟化合物具有有吸引力的化学、药理学和生物学特性，使他们能够在生物制药和生物材料的设计中进行范式转变。将氟原子引入氨基酸和核酸中，开辟了广阔的新化学领域改变肽、蛋白质和 DNA 的折叠、稳定性、寡聚倾向和生物活性。然而，尽管显示出具有良好的特性，但在生物支架中添加氟的影响是此外，越来越多的证据表明全氟化合物会混杂地吸附。细胞的许多基本组成部分 - 包括脂质、蛋白质和 DNA - 可以引发多种 PI 实验室最近发现的这些效应之一是有机氟分子的能力。直接将蛋白质和 DNA 组装成含氟微结构域，然后相分离成氟化液体，无需 PI 最近利用这些新特性来实现超声引导。基于这些初步发现，提出了三维组织中氟化蛋白质的研究。研究计划将机械地探索有机氟化合物如何影响结构和吸附蛋白质和 DNA 的功能，并利用这些见解来指导新型超分子的设计我们的总体假设是有机氟化合物非共价吸附。蛋白质和 DNA 引导它们分离成富氟相，这反过来又改变了它们的寡聚体为了检验这一论断，我们将扩展我们的全氟化合物 (PFC)。我们将使用包含具有碱性/酸性官能团和杂环部分的多种分子的库。该库可建立控制 PFC 吸附蛋白质能力的结构-活性关系，以及研究 PFC 络合如何改变蛋白质细胞摄取、细胞内运输和生物活性。同时，我们将使用这个库来研究介导 PFC-DNA 相互作用的分子机制并检查这些研究共同探讨了 PFC 络合如何改变暴露细胞中的 DNA 稳定性和代谢稳态。将建立对 PFC 如何与蛋白质和 DNA 相互作用的全面机制理解，并将让我们能够合理地设计氟生物技术，利用不寻常的组装现象和相举例来说，我们将创建超声波敏感的氟纳米乳液。装载有 PFC 修饰的核糖核蛋白 (RNP)，可在肾脏组织中进行成像引导的基因编辑。这项研究的成功将推动 PFC 作为控制蛋白质和 DNA 的新分子基序的使用组装，以及开发的方法用于发现细胞内转导的新试剂最终，提高有关有机氟化合物如何相互作用的知识。蛋白质和 DNA 及其对细胞的影响，将指导新型 PFC 技术的合理设计具有药物发现和纳米医学应用所需的功能特性。