Exploiting gold nanoparticle as a probe to identify therapeutic targets

纳米颗粒(NPs)主要用作各种生物医学应用的载体。什么时候 在生物流体中，NPs与蛋白质相互作用，在其表面形成一层生物涂层，称为蛋白质 科罗娜。已经对NPs周围的蛋白质电晕进行了研究，以解决包括 NP的生物分布、清除和潜在毒性。此前，我们和其他人一起展示了自我 金纳米颗粒(GNPs)的治疗特性。在目前的应用中，开发自疗性GNP (ST-GNP)作为一个探针，我们提出了一个独特的概念来捕获、识别和验证治疗 负责肿瘤生长和癌症治疗阻力的靶点。 我们证明ST-GNP抑制了一些促肿瘤肝素结合的功能 通过结合HB结构域改变蛋白质构象的生长因子(HB-GFS)，而 非Hb-GFS的构象和功能保持不变。此外，在各种规模中，国民生产总值为20% 纳米粒显示出最高的治疗效果，而100 nm粒的GNP是非自疗的(NST- 国民生产总值)。重要的是，ST-GNP抑制肿瘤的生长、转移和增敏卵巢癌细胞对顺铂的敏感性 逆转原位上皮-间充质转化(EMT)并取消MAPK信号转导(图1) 在胰腺癌模型上，我们报道了ST-GNP干扰癌细胞和癌症之间的串扰 抑制肿瘤生长的相关成纤维细胞(CAF)和重新编程的肿瘤微环境。 在研究卵巢癌或正常细胞裂解液中ST-GNP的蛋白质浓缩时，我们发现 SMNDC1和PPA1是潜在的肿瘤生长新靶点。此外，我们最近报道了无毒的 Auroliposome增强siRNA沉默效果并更有效地抑制卵巢肿瘤生长 与传统的以DOTAP-DOPE为基础的脂质体传递siRNA相比。基于这些结果，我们 假设富含ST-GNP的蛋白质的功能将受到抑制，从而导致肿瘤生长 抑制和治疗耐药。因此，这些富含ST-GNP的蛋白可能具有潜在的治疗作用 目标。我们将使用下面的具体目标来检验我们的假设； 具体目的1：研究ST-GNP对蛋白质的富集性。 具体目标2：在动物模型中验证治疗靶点。 影响：NPs周围的蛋白质日冕正在演变为个性化药物的独特标志。我们的发现 支持ST-GNP不仅用于确定卵巢和胰腺的治疗靶点 癌症，但在糖尿病视网膜病变、黄斑变性和类风湿性关节炎中也有 报道了GNP的ST特性在这些模型中抑制血管生成。

Nanoparticles (NPs) have mostly been used as delivery vehicles for various biomedical applications. When exposed to biological fluids NPs interact with proteins forming a biological coating on their surface, termed protein corona. Protein corona around NPs have been investigated to address the biological responses including biodistribution, clearance and potential toxicity of NP. Previously, we along with others have demonstrated self- therapeutic property of gold nanoparticles (GNPs). In the current application, exploiting self-therapeutic GNP (ST-GNP) as a probe, we are proposing a unique concept of capturing, identifying and validating therapeutic targets responsible for tumor growth and therapy resistance in cancer. We demonstrated that ST-GNP inhibited functions of a number of tumor-promoting heparin-binding growth factors (HB-GFs) via binding through the HB-domain that altered protein conformations, whereas conformations and functions of non-HB-GFs remained unaltered. In addition, among various sizes, GNP of 20 nm size demonstrated highest therapeutic efficacy whereas GNP of 100 nm size was non self-therapeutic (NST- GNP). Importantly, ST-GNP inhibited tumor growth, metastasis and sensitized ovarian cancer cells to cisplatin by reversing epithelial-mesenchymal transition (EMT) and abrogating MAPK-signaling (Fig 1) In orthotopic model of pancreatic cancer, we reported that ST-GNP disrupted cross-talk between cancer cells and cancer associated fibroblasts (CAFs) and reprogrammed tumor microenvironment that inhibited tumor growth. Investigating protein enrichment on ST-GNP from ovarian cancer or normal cellular lysates, we identified SMNDC1 and PPA1, as potential new targets for tumor growth. Furthermore, we recently reported that non-toxic Auroliposome enhanced silencing efficacy of siRNA and more effectively inhibited ovarian tumor growth compared to traditional DOTAP-DOPE based liposomal delivery of siRNA. Based on these results, we hypothesize that functions of the proteins enriched on ST-GNP will be inhibited resulting in tumor growth inhibition and therapy resistance. Therefore, these ST-GNP-enriched proteins may serve as potential therapeutic targets. We will use specific aims below to test our hypothesis; Specific aim 1: Investigating protein enrichment on ST-GNP. Specific aim 2: Validating therapeutic targets in animal models. Impact: Protein corona around NPs is evolving as a unique signature for personalized medicine. Our findings support that the ST-GNP could be utilized to identify therapeutic targets not only for ovarian and pancreatic cancer but in diabetic retinopathy, macular degeneration and rheumatoid arthritis as well where others have reported ST property of GNP to inhibit angiogenesis in these models.