Name: analysis of targeted viral protein nanoparticles delivered to her2+ tumors
Uploaded: 2013-06-18T12:00:00
Description: Watch this Scientific Journal Video about Analysis of Targeted Viral Protein Nanoparticles Delivered to HER2+ Tumors at JoVE.com

这项工作是由美国国立卫生研究院/美国国家癌症研究所（R01CA129822 R01CA140995）LKM-K补助。 Medina博士Kauwe感谢C.国王，M. M-Kauwe D. Revetto的的继续支持。

1。全身交付体内的 <ol><li>用无菌生理盐水足够HerDox的混合等同于0.2毫升0.004毫克/千克的剂量注射HerDox每6-8周的老NU / NU鼠标轴承皮下双边侧翼异种移植肿瘤。 </li><li>轻轻绘制HerDox的混合物倒入一个3/10毫升配备29G针头的胰岛素注射器，避免气泡。 </li><li>进行了简短的异氟醚麻醉诱导感应室配备一个气体净化系统（氧气流速：0.5-1升/分钟，异氟醚浓度：3-4％（或更低）。 </li><li>整个混合物?...

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阿霉素荧光可以检测到体内时使用多模成像肿瘤皮下。然而，治疗有效剂量的HerDox（0.004毫克/千克）的检测阈值以下后，单次剂量。相比之下，经过7每天注射（7天1x/day），，肿瘤的积累和保留的粒子是足以使阿霉素荧光可视化。 这是至关重要的工作时，与阿霉素在体内成像，使用清洁技术或任何其他荧光。鼠标的外侧上的滴水应避免，因为成像器将检测到皮肤...

肿瘤靶向化疗有可能消除癌细胞不相关的药物相比，在​​较低的剂量，因为可以在其预定的目的地，而不是分发非肿瘤组织蓄积的交付治疗。作为后一种情况下，会稀释药的药效，因此需要较高的剂量是有效的，肿瘤靶向治疗和安全性优于标准的非针对性的治疗。 通过封装在自组装纳米颗粒靶向化疗使药物保持化学改性的共价连接到靶向分子的药物相比。因此联动有可能改变的活性的药物，靶向分子非共价键的组件允许保留药物效力。 我们先前的研究显示，这种新型三分量，复杂的自组装，HerDox，针对HER2 +肿瘤<em在体内诱发肿瘤生长消融，同时保留正常组织，包括心脏1。通过受体结合的细胞渗透蛋白，HerPBK10，与化疗剂之间的非共价相互作用，形成HerDox，多柔比星（阿霉素），通过一个小的核酸连接器。 HerPBK10结合人类表皮生长因子受体（HER）和，触发受体介导的内吞作用2-4，而核内体膜的渗透是通过腺病毒衍生的聚氯基地衣壳蛋白4-6成立。甲带正电荷的蛋白质域使核酸结合4，5，通过该DNA插霉素可以用于靶向递送运。亲电，插层，并有可能蛋白低聚相互作用促进自组装成圆形10-20nm的颗粒是稳定的血液和根据扩展存储在不同温度下1。她的优先目标2 +肿瘤细胞是促进增强配体的亲和力，时HER2升高。 我们以前的研究已经表明，在对非肿瘤组织中的肿瘤相比，优惠HerDox产量积累全身给药不相关的Dox诱导1，渗透进入肿瘤细胞在体内 7。我们已经观察到HerDox释放Dox诱导肿瘤细胞的条目后，允许进入细胞核1的阿霉素积累。肿瘤积累与受体水平相对较低HER2表达肿瘤积累减HerDox相比相对较高的HER2水平1。此外，有效的细胞死亡的浓度呈现负相关与HER2显示肿瘤细胞株表达不同的细胞表面HER2水平。 HerDox具有不相关的DOX治疗和安全的优势，作为肿瘤杀伤作用发生在超过10倍，低剂量相比解压geted药物和产生心（通过超声心动图和组织学染色检测）或肝功能（TUNEL检测染色）组织上没有检测的不利影响，相反，没有针对性霉素1。尽管其从病毒衣壳蛋白的推导，HerPBK10展品没有检测免疫原性治疗水平2。鉴于整个腺病毒的预先存在的抗体可以识别HerPBK10，他们是无法防止细胞结合2。 测量肿瘤体积随着时间的推移是一个标准的方法，靶向治疗的疗效评估，并已为评估疗效HerDox的雇用。补充这种方法在体内和体外荧光强度成像已经让我们更好地评估靶向效率7。我们已经专门集成在切除肿瘤的原位共焦成像光谱的DOX荧光分析来验证HerDox只有积累在体内的肿瘤，但渗透到肿瘤细胞和交付阿霉素进入细胞质和细胞核7。此外，频谱分析使我们能够区分霉素荧光自发荧光7。 这里，我们证明我们的方法更详细地用于全身给药后在体内的评估HerDox的，最重要的是，通过多模成像方法和分析评估目标。

<table border="1">
<tbody>
 <tr>
 <td>Name of the reagent</td>
 <td>Company</td>
 <td>Catalogue number</td>
 <td>Comments (optional)</th
 ></tr>
 <tr>
 <td >Fluorescence laser scanning confocal microscope</td>
 <td >Leica</td>
 <td >SPE</td>
 <td></td>
 </tr>
 <tr>
 <td>In Vivo Optical Imager</td>
 <td>Spectral Molecular Imaging</td>
 <td>Multimode In Vivo Optical Imager</td>
 <td></td>
 </tr>
 <tr>
 <td >Doxorubicin-HCl</td>
 <td >Sigma-Aldrich</td>
 <td >D4035</td>
 <td></td>
 </tr>
 <tr>
 <td >Nude (NU/NU) mouse, female, 6-8 week</td>
 <td >Charles River</td>
 <td >Strain code 088</td>
 <td></td>
 </tr>
 <tr>
 <td >MDA-MB-435 human HER2+ tumor cells</td>
 <td >NCI-Frederick Cancer DCTD Tumor/Cell Line Repository</td>
 <td >0507292</td>
 <td></td>
 </tr>
 <tr>
 <td >3/10 cc insulin syringe U-100 with 29G x 1/2" Ultra-FineIV permanently attached needle</td>
 <td >BD</td>
 <td >309301</td>
 <td></td>
 </tr>
 <tr>
 <td >Delta T chamber </td>
 <td >Bioptechs </td>
 <td >04200417B</td>
 <td></td>
 </tr>
 </tbody>
</table>

analysis of targeted viral protein nanoparticles delivered to her2+ tumors

HER2 +肿瘤靶向纳米粒子，HerDox，具有肿瘤优惠的积累和消融治疗HER2 +癌症动物模型中肿瘤生长。 HerDox形成通过非共价的肿瘤靶向与化疗剂，多柔比星的细胞渗透蛋白的自组装体，通过一个小的核酸连接器。电，插层，齐聚相互作用的结合，促进成圆形的10-20纳米粒子自组装。 HerDox显示出在不同温度下的血中稳定性，以及在扩展存储。全身给药HerDox在荷瘤小鼠的肿瘤细胞死亡的结果没有检测到不利影响的非肿瘤组织，包括心脏和肝脏（发生明显损坏由无目标的多柔比星）。 HER2海拔细胞中的表达相比，有利于靶向表达人表皮生长因子受体的细胞，因此，显示升高的HER2水平的肿瘤表现出较大的积累HerDox唱较低的水平，无论是在体外和体内 。 原位共焦和频谱分析相结合的荧光强度成像已经让我们在体内肿瘤靶向肿瘤细胞渗透全身给药后HerDox的验证。在这里，我们详细介绍我们的方法，以评估肿瘤靶向全身给药后通过多模成像。

图1显示了在体内光学成像样机下图像采集多种方式，包括荧光强度，光谱，寿命，双光子，重要的内焦，生物发光成像的目的，这是建。此外，冷却的高灵敏度的摄像头和纳入在本系统中的高功率激光线会产生更高的对比度比商业光学成像系统11的荧光图像，特别是用于检测体内阿霉素荧光。因此，该系统是用于在本研究中，获得多个互补的数据点在体内</em...

Watch this Scientific Journal Video about Analysis of Targeted Viral Protein Nanoparticles Delivered to HER2+ Tumors at JoVE.com

Analysis of Targeted Viral Protein Nanoparticles Delivered to HER2+ Tumors

本文详细介绍了肿瘤靶向纳米粒子，HerDox光学成像分析的程序。特别是，使用多模成像装置，用于检测肿瘤靶向和评估肿瘤渗透详细描述在这里。

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