Lipid Nanoparticle Mediated Inhibition and Detection of Matrix Metalloproteinases

描述(申请人提供)：脂质体是脂基纳米粒的最好代表之一，由于其易于配制以及生物兼容性，它们非常适合作为药物输送载体。除了药物载体外，脂质体纳米颗粒还具有形成针对蛋白质的模板表面的潜力(由于其脂质单体的固有流动性)，并且这种特征可以被设计用于开发具有生物医学意义的蛋白质的高度特异性诊断和脱敏方案。这项研究将开发基于脂质体的新型人工抗体，它将在同工酶特异性检测和抑制四种基质金属蛋白酶(即，MMP2、MMP7、MMP9和MMP10)方面得到应用，这四种MMP2、MMP7、MMP9和MMP10已知与多种人类疾病的发病机制有关。这些基础的、过程开发的研究将在以下特定目标下完成：(I)合成作为MMPs的初始锚点的脂质结合物，以及蛋白质表面的相互作用。(Ii)在每种MMPs(即，MMP2、-7、-9和-10)存在下的脂质体的模板聚合，并测试所得到的(聚合的)脂质体对其母MMPs的同工酶选择性抑制的潜力。(Iii)利用脂质体包裹的稀土离子的时间分辨发光光谱对MMPs进行同工酶选择性检测。(4)蛋白质-脂质相互作用在同工酶选择性检测和抑制MMPs中作用的机理研究。这些研究目标将通过使用合成有机化学、分子生物学、荧光光谱以及酶-配体相互作用和酶催化的动力学和热力学分析技术来实现。这项研究的成功完成将为在不同的病理生理条件下开发基于脂质体纳米颗粒的高特异性和简便性的其他病原体和生物标志物蛋白的诊断工具提供见解。公共卫生相关性：拟议的研究结果将在诊断和治疗各种癌症方面找到长期应用。在此应用期间，我们将集中精力设计新的脂质纳米颗粒，并了解检测和抑制参与各种癌症进展和转移的几种酶所固有的基本机制原理。这项研究的成功完成将为开发其他导致人类不同疾病的蛋白质的诊断和治疗工具奠定基础。

DESCRIPTION (provided by applicant): Liposomes are one of the best representatives of lipid-based nanoparticles, and they are ideally suited as the drug delivery vehicles due to their ease in formulations as well as biocompatibility. Aside from drug carriers, the liposomal nanoparticles have potentials to form template surfaces against proteins (due to intrinsic mobility of their lipid monomers), and such features can be contrived to develop highly specific diagnostic and desensitization protocols for proteins of biomedical relevance. The proposed research will develop novel liposome-based artificial antibodies , which will find applications in isozyme specific detection and inhibition of four matrix metalloproteinases (viz., MMP-2, MMP-7, MMP-9, and MMP-10), which are known to be involved in the pathogenesis of a variety of human diseases. These fundamental, process developmental , research will be accomplished under the following specific aims: (i) Synthesis of lipid conjugates to serve as the initial anchor sites for MMPs, as well as for interaction at the protein surfaces. (ii) Template polymerization of liposomes in the presence of each of the MMPs (i.e., MMP-2, -7, -9 and -10), and testing the potentials of resultant (polymerized) liposomes in isozyme selective inhibition of their parent MMPs. (iii) Isozyme-selective detection of the MMPs by employing time-resolved luminescence spectroscopy of liposome-incorporated lanthanide ions. (iv) Mechanistic studies on the roles of protein-lipid interactions in isozyme-selective detection and inhibition of MMPs. These research objectives will be accomplished by employing the techniques of synthetic organic chemistry, molecular biology, fluorescence spectroscopy, and kinetic and thermodynamic analyses of enzyme-ligand interactions and enzyme catalyses. The successful completion of this research will provide insights into developing liposomal nanoparticle-based highly specific and facile diagnostic tools for other pathogenic as well as biomarker proteins under diverse pathophysiological conditions. PUBLIC HEALTH RELEVANCE: The outcome of the proposed research will find long term applications in diagnosing and treating various cancers. During this application period, we will focus our attention in designing novel lipid nanoparticles, and in understanding the basic mechanistic principles intrinsic to the detection and inhibition of several enzymes involved in the progression and metastasis of various cancers. The successful completion of this research will establish foundation for developing diagnostic and therapeutic tools for other proteins responsible for causing different human diseases.