Theranostic Nanoparticles to enhance morpholino delivery to the liver for suppres

治疗诊断纳米颗粒可增强吗啉向肝脏的输送以抑制

• 批准号: 8490376
• 负责人: David Peter Cormode
• 金额: $ 22.86万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8490376
• 关键词: Applications Grants; Attention; Award; Biodistribution; Biological; Blood; Blood Circulation; Cardiomyopathies; Cells; China; Cholesterol; Cleaved cell; Contrast Media; Cytoplasm; DNA delivery; Detection; Development; Developmental Biology; Disease; Endosomes; Fluorescence; Galactose; Gene Expression; Gene Silencing; Genetic Materials; Goals; Grant; Half-Life; Health; Heart Diseases; Hepatocyte; Hypertrophy; Imaging Techniques; In Vitro; Incidence; Individual; Injection of therapeutic agent; Investigation; Iron; Label; Lead; Liver; Magnetic Resonance Imaging; Malignant Neoplasms; Mediating; Medical Imaging; Mentors; Messenger RNA; Methods; MicroRNAs; Mus; Muscle; Myocardial Infarction; Northern Blotting; Nucleic Acids; Organ; Outcome; Outcome Measure; Patients; Performance; Pharmaceutical Preparations; Phase; Polyethylene Glycols; Polymers; Population; Production; Property; Proprotein Convertase 1; Proprotein Convertases; Publications; Quality of life; RNA; RNA Splicing; Reporting; Residual state; Resistance; Role; Rupture; Serum; Site; Small Interfering RNA; Specificity; Staging; Stenosis; Subtilisins; Surface; System; Techniques; Technology; Testing; Therapeutic; Tissues; Toxic effect; Training; Viral; Western Blotting; Work; base; cell type; dithiol; experience; fluorophore; gene therapy; heart disease risk; human disease; improved; in vitro testing; in vivo; iron oxide; kexin; knock-down; medical schools; nanoparticle; novel; nuclease; particle; pre-clinical; research study; scaffold; success; theranostics; uptake

ABSTRACT Gene therapy holds great promise for new treatments for many diseases. Despite great pre-clinical successes, few gene therapy treatments have been effective in patients. In this grant application, is proposed a new type of gene therapy system, namely morpholino-nanoparticles. Morpholinos are highly effective for suppressing gene expression and, notably, for suppressing microRNA function. The nanoparticles used will possess the following features: morpholinos attached using a dithiol bond that will be cleaved in endosomes; a polymer coating that can disrupt endosomes for morpholino release into the cytoplasm; PEG chains to allow a long circulation half-life; galactose targeting to hepatocytes; an iron core for MRI and TEM detection; and a fluorophore for fluorescence techniques. The iron oxide and fluorophore components of the nanoparticle allow for a ¿theranostic¿ approach, where the nanoparticle performance can be evaluated using imaging techniques. Due to the long circulation half-life of these nanoparticles and the galactose targeting, the nanoparticles should localize in the hepatocytes of the liver in vivo. Two approaches for reducing cholesterol production will be attempted: 1) PCSK9 knockdown and 2) miR-122 suppression. Cholesterol levels are correlated with the risk of heart disease and therefore this morpholino-nanoparticle system would be a treatment for individuals with elevated levels of cholesterol. However, the morpholino-nanoparticle delivery technology developed under this grant could subsequently be applied for therapy of other aspects of heart disease such as hypertrophy, cardiomyopathy and stenosis, or to other diseases such as cancer. The candidate is highly experienced in the synthesis of multifunctional nanoparticles that act as targeted contrast agents for medical imaging. The purpose of this award is to train the candidate to develop and apply novel nanoparticles for gene therapy purposes. The K99 mentored phase of the award will take place under the guidance of Prof. Roger Hajjar and Prof. Zahi Fayad of Mount Sinai School of Medicine. The focus of the training will be on the development of nanoparticles for gene therapy and the techniques required for analysis of mRNA and microRNA knockdown, i.e. PCR, Western blotting and Northern blotting. This mentored phase will set the stage for the R00 independent phase where the nanoparticle development and in vitro testing will continue and in vivo trials will be initiated. The results of this work should lead to the establishment of a fruitful line of investigation for the candidate that will reap benefits for our understanding of disease and human health.

抽象的 基因疗法对许多疾病的新疗法具有很大的希望。尽管临床前的成功很大， 很少有基因治疗对患者有效。在此赠款申请中，提出了一种新类型 基因治疗系统，即形态纳米颗粒。吗啡对抑制非常有效 基因表达，特别是用于抑制microRNA功能。所使用的纳米颗粒将拥有 以下特征：使用将在内体中切割的二硫醇键附着的吗啡；聚合物 可能破坏内体的涂层，以释放到细胞质中；钉链允许长时间 圈子半衰期；半乳糖靶向肝细胞； MRI和TEM检测的铁芯；和 荧光团用于荧光技术。纳米颗粒的氧化铁和荧光团成分允许 对于«theranostic？，可以使用成像技术评估纳米颗粒性能。 由于这些纳米颗粒和半乳糖靶向的长圆半衰期，纳米颗粒应 位于体内肝细胞中。减少胆固醇产生的两种方法将是 尝试：1）PCSK9敲低和2）miR-122抑制。胆固醇水平与 心脏病，因此该形态纳米颗粒系统将是针对患有 胆固醇水平升高。但是，在此基础上开发的吗啡 - 纳米颗粒输送技术 随后，Grant可以用于治疗心脏病的其他方面，例如肥大， 心肌病和狭窄，或其他疾病，例如癌症。 候选人在合成多功能纳米颗粒方面经验丰富，这些纳米颗粒的作用 医学成像的对比剂。该奖项的目的是培训候选人开发和申请 用于基因治疗目的的新型纳米颗粒。 奖项的K99修订阶段将在罗杰·哈哈尔（Roger Hajjar）教授和扎希教授的指导下举行。 西奈山医学院的法亚德。培训的重点将放在纳米颗粒的开发上 用于基因疗法以及分析mRNA和microRNA敲低所需的技术，即PCR， 蛋白质印迹和北印迹。这个修补阶段将为R00独立阶段奠定阶段 纳米颗粒发育和体外测试将继续进行，并开始体内试验。这 这项工作的结果应导致为候选人建立富有成果的投资线 收获我们对疾病和人类健康的理解。