TRACEABLE CANCER-TARGETED NANOPARTICLES FOR LOCALIZED CHEMOTHERAPY

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。纳米技术为开发特定部位化疗开辟了新的途径。我们正在设计携带化疗药物或磁共振成像造影剂的纳米颗粒，分别用于癌症治疗和诊断。研究表明，由于未成熟、渗漏的血管系统，纳米颗粒可以选择性地在肿瘤中积聚。我们的目标是通过在纳米颗粒表面添加靶向多肽来增加这种选择性积累。到目前为止，我们的研究主要集中在制备可生物降解的聚(dl-乳酸-羟基乙酸)(PLGA)(PLGA)的~50、~250和~500 nm的均一纳米粒子。已证实环肽cLABL与细胞间黏附分子-1(ICAM-1)具有高亲和力，细胞内黏附分子-1在肺癌、胰腺癌、前列腺癌等中表达上调。我们将cLABL固定在PLGA纳米颗粒的表面，并通过ThioGlo荧光分析检测多肽上是否存在二硫键来定量表面密度。为了增加cLABL与ICAM-1在细胞表面的可及性，我们将碳二亚胺或聚乙二醇间隔物连接到纳米颗粒表面，然后将cLABL与这些间隔物的末端反应。在一项平行的研究中，我们与堪萨斯大学医学中心霍格伦德脑成像中心的Mehmet Bilgen博士合作，测试我们将Gd封装到PLGA纳米颗粒中作为MRI对比剂的能力。他已经验证了这些纳米颗粒配方以浓度依赖的方式增强磁共振图像的能力。在正在进行的研究中，我们正在制造连接和不连接cLABL的荧光纳米颗粒。这些制剂将与癌细胞Caco-2或内皮细胞单层(HUVECs)孵育，以量化cLABL纳米颗粒与未标记纳米颗粒的粘附性或内吞作用。同时，我们计划通过核磁共振比较cLABL-纳米颗粒和未标记纳米颗粒的生物分布。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Nanotechnology has opened new avenues toward the development of site specific chemotherapy. We are designing nanoparticles that carry either a chemotherapeutic or a contrast agent for magnetic resonance imaging for application in cancer treatment and diagnosis, respectively. Nanometer-sized particles have been shown to selectively accumulate in tumors due to immature, leaky vasculature. Our goal is to augment this selective accumulation by adding a targeting peptide to the nanoparticle surface. Our studies to date have focused on fabricating homogeneous populations of ~50, ~250, and ~500 nm nanoparticles of biodegradable poly(dl-lactic-co-glycolic acid) (PLGA). The cyclic peptide, cLABL, has been proven to bind with high affinity specificity to intracellular cell adhesion molecule-1 (ICAM-1), which is upregulated in lung cancer, pancreatic cancer, prostate cancer, etc. We have tethered cLABL to the surface of PLGA nanoparticles and quantified the surface density by a detecting the presence of the di-sulfide bond on the peptide via a ThioGlo fluorescence assay. To increase the accessibility of cLABL to ICAM-1 on the surface of cells, we have attached carbodiimide or polyethylene glycol spacers to the nanoparticle surface and subsequently reacted cLABL to the distal end of these spacers. In a parallel study, we have partnered with Dr. Mehmet Bilgen at the Hoglund Brain Imaging Center at the University of Kansas Medical Center to test our ability to encapsulate gadolinium into PLGA nanoparticles as an MRI contrast enhancing agent. He has verified the ability of these nanoparticle formulations to enhance magnetic resonance images in a concentration dependent manner. In ongoing studies, we are fabricating fluorescent nanoparticles with and without cLABL attached. These formulations will be incubated with cancerous Caco-2 or endothelial cell monolayers (HUVECs) to quantify the adhesion or endocytosis of cLABL-nanoparticles compared to unlabeled nanoparticles. In parallel, we plan to compare the biodistribution of cLABL-nanoparticles to unlabeled nanoparticles via MRI.