Modular Nanomedicines Based on Heterogeneous Fusion Protein Co-Assembly

基于异质融合蛋白共组装的模块化纳米药物

• 批准号: 9145217
• 负责人: Gregory Hudalla
• 金额: $ 7.03万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-16 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9145217
• 关键词: Address; Adjuvant; Antigens; Architecture; Binding; Biocompatible; Cells; Chimeric Proteins; Clinical; Complex; Construction Materials; Development; Diagnostic; Disease; Dose; Drug Delivery Systems; Electrostatics; Engineering; Family; Genes; Genetic Recombination; Goals; Health; Immune response; In Vitro; Lead; Leucine Zippers; Ligands; Link; Mediating; Mono-S; Mus; Names; Nature; Peptides; Pharmaceutical Preparations; Pre-Clinical Model; Process; Property; Protein Engineering; Protein Subunits; Proteins; Recombinant Proteins; Recombinants; Research; Serum; Site; Structure; Techniques; Tertiary Protein Structure; Testing; Therapeutic; Toxic effect; Work; base; controlled release; design; flexibility; fusion gene; immunogenic; immunogenicity; innovation; interest; microbial host; molecular assembly/self assembly; monomer; mutant; nanoassembly; nanofiber; nanomaterials; nanomedicine; nanoscale; programs; self assembly; small molecule; success; therapeutic protein; therapeutic target

 DESCRIPTION: Recombinant proteins are promising as therapeutics because they have specific functional properties that often cannot be adequately mimicked by small molecule drugs. However, the inability of many therapeutic proteins to concentrate at a disease site typically necessitates administration at relatively high doses, which often results in off-site effects and toxicity. Numerous protein engineering and controlled-release strategies have been investigated to enhance site-specific protein delivery, however, each is hindered by different mechanisms that can diminish protein activity. The primary goal of the proposed research is to create a facile approach to endow therapeutic proteins with targeting properties, without requiring a drug delivery vehicle or extensive protein engineering. Throughout nature, self-assembly of different functional proteins gives rise to supramolecular entities that can perform complex tasks via the co-integrated activity of each protein subunit. The long-term goal of our research program is to create multifunctional nanomedicines with precisely defined composition of therapeutic, targeting or diagnostic properties via self-assembly of different engineered proteins. To achieve this goal, the proposed research will create a trio of engineered peptide "tags" that selectively co-assemble into a heterogeneous a-helical coiled-coil via electrostatic and hydrophobic complementarity. Multifunctional nanomaterials will then be created through a three-step process: 1) genes encoding each of these peptide tags will be linked to genes encoding different functional protein ligands, 2) these recombinant fusion genes will be expressed by microbial hosts, and 3) the recovered fusion proteins will co-assemble into a precisely defined tripartite nanomaterial having function related to each co-integrated ligand upon mixing. Importantly, because the recombinant fusion tags mediating assembly are independent from the protein ligand itself, we envision that the functional protein component can be easily interchanged via DNA recombination, thus allowing for functional properties of the resulting nanomaterial to be precisely varied without altering its assembly. In Specific Aim 1, we will develop fusion proteins that assemble into a trifunctional nanomaterial via engineered peptide domains that selectively assemble into a heterogeneous trimeric a-helical coiled-coil. We will characterize influence of co-assembly on the bioactivity of each integrated ligand, as well as the serum stability of these assemblies. In Specific Aim 2, we will then assess the immunogenicity of peptide and fusion protein assemblies based on heterogeneous a-helical coiled-coils, expecting that they will be minimally immunogenic in the absence of an immunostimulatory adjuvant. Success of the proposed research will add significantly to the growing body of work utilizing a-helical coiled-coils to create multimeric protein nanoassemblies, by providing a general approach for modular nanomaterial design. In turn, these materials may provide the basis for nanomedicines with modular composition of therapeutic, targeting and diagnostic that can be precisely tailored to address a broad spectrum of diseases.

 描述：重组蛋白作为治疗药物很有希望，因为它们具有小分子药物往往无法充分模仿的特殊功能特性。然而，由于许多治疗性蛋白质不能集中在疾病部位，通常需要相对较高的剂量给药，这往往会导致非现场效应和毒性。许多蛋白质工程和控释策略已经被研究以增强特定部位的蛋白质传递，然而，每种策略都受到不同的机制的阻碍，这些机制可以降低蛋白质的活性。这项拟议研究的主要目标是创造一种简便的方法来赋予治疗性蛋白质靶向特性，而不需要药物输送载体或广泛的蛋白质工程。在自然界中，不同功能蛋白质的自组装产生了超分子实体，这些超分子实体可以通过每个蛋白质亚单位的共同整合活性来执行复杂的任务。我们研究计划的长期目标是通过不同工程蛋白的自组装，创造具有精确定义的治疗、靶向或诊断特性组成的多功能纳米药物。为了实现这一目标，这项拟议的研究将创造三个工程化的多肽“标签”，它们通过静电和疏水互补选择性地共同组装成一个异质的a-螺旋螺旋线圈。然后将通过三个步骤创建多功能纳米材料：1)编码每个肽标签的基因将连接到编码不同功能蛋白配体的基因，2)这些重组融合基因将由微生物宿主表达，以及3)恢复的融合蛋白将在混合时共同组装成具有与每个共整合配体相关的功能的精确定义的三部分纳米材料。重要的是，由于重组融合标签介导的组装独立于蛋白质配体本身，我们设想功能蛋白质组分可以通过DNA重组很容易地互换，从而允许得到的纳米材料的功能特性在不改变其组装的情况下精确地改变。在具体目标1中，我们将开发融合蛋白，通过工程多肽结构域选择性地组装成异质三聚体a螺旋螺旋卷曲，从而组装成三功能纳米材料。我们将表征共组装对每个整合配体的生物活性的影响，以及这些组装的血清稳定性。在特定的目标2中，我们将评估基于异质a螺旋螺旋线圈的多肽和融合蛋白组件的免疫原性，期望在没有免疫刺激佐剂的情况下，它们将是最低限度的免疫原性。这项拟议研究的成功将通过提供模块化纳米材料设计的一般方法，显著增加利用a螺旋螺旋线圈创建多聚体蛋白质纳米组件的工作总量。反过来，这些材料可能为纳米医学提供基础，这些纳米医学具有治疗、靶向和诊断的模块化组合，可以精确地定制以解决广泛的疾病。