溶瘤病毒的Transarterial管理局大鼠原位肝癌的局部治疗
Summary
Oncolytic virotherapies are under development as novel therapeutics for the treatment of hepatocellular carcinoma (HCC). Here we describe a method for locoregional therapy of HCC via hepatic arterial administration of oncolytic virus.
Abstract
Hepatocellular carcinoma (HCC) is a disease with limited treatment options and poor prognosis. In recent years, oncolytic virotherapies have proven themselves to be potentially powerful tools to fight malignancy. Due to the unique dual blood supply in the liver, it is possible to apply therapies locally to orthotopic liver tumors, which are predominantly fed by arterial blood flow. We have previously demonstrated that hepatic arterial delivery of oncolytic viruses results in safe and efficient transduction efficiency of multifocal HCC lesions, resulting in significant prolongation of survival in immune competent rats. This procedure closely mimics the application of transarterial embolization in patients, which is the standard palliative care provided to many HCC patients. The ability to administer tumor therapies through the hepatic artery in rats allows for a highly sophisticated preclinical model for evaluating novel viral vectors under development. Here we describe the detailed protocol for microdissection of the hepatic artery for infusion of oncolytic virus vectors to treat orthotopic HCC.
Introduction
肝细胞癌（HCC）是世界上第五最常见的癌症，和与癌症有关的死亡的第三大原因，使其成为一个显著健康关注1,2。对于谁是病人没有资格肿瘤切除术，或那些等待肝移植，局部治疗包括经动脉栓塞（TAE）或肝动脉化疗栓塞（TACE）应用于标准姑息治疗3,4。这些疗法利用在肝脏肿瘤即是由肝动脉血流几乎完全喂双重血供的独特功能，而周围的肝脏受到多数来自门静脉5,6其供血。
由于肝癌的治疗确立的极为有限的功效，溶瘤病毒已成为有前途的替代疗法。 JX-594，最近改名为Pexa-VEC，是胸苷激酶缺失牛痘病毒，具有粒米武装acrophage细胞集落刺激因子（GM-CSF），它已经完成了肝癌7 II期临床试验。最近，表达人干扰素-β的重组水泡性口炎病毒载体（VSV）已进入I期临床试验为索拉非尼耐火肝癌（NCT01628640）。作为溶瘤病毒靠拢经批准用于患者的临床应用为肝细胞癌，需要一个有效的给药途径对目标多焦点疾病是显而易见的。而全身性递送是很大程度上是无效的，由于低效肿瘤转导，瘤内的应用程序可能会限制疗法所注入的肿瘤的功效，留下uninjectable微观病变易患疾病的进展。
我们已经建立了分离大鼠肝动脉施用溶瘤病毒疗法在一个局部的方式定位到原位HCC的方法。我们已经证明，这种给药途径导致安全和effecti已经多焦点肝癌结节转导，导致免疫活性大鼠8-10显著存活延长。这里，我们描述访问，解剖，并注入大鼠肝动脉的方法。该过程的图解示于图1（先前公布9）
Protocol
Representative Results
Discussion
Although direct intratumoral injection is undoubtedly the simplest method to result in efficient tumor transduction of a single tumor nodule, hepatic arterial infusion represents an ideal administration route to target multifocal, orthotopic HCC. This method has proven to be both safe and effective for treating HCC in immune competent rats with oncolytic viruses. Furthermore, since HCC patients are routinely treated by transarterial application of chemoembolization, the method described here is readily translatable to …
Disclosures
The authors have nothing to disclose.
Acknowledgements
This work is supported by the SFB 824 subprojects C6 and C7 (DFG Sonderforschungsbereich 824), German Research Foundation, Bonn, Germany.
Materials
| Veterinary clippers | Aesculap | GT415 | Small, cordless trimmer ideal for removing fur from surgical area |
| Stereomicroscope | Zeiss | Stemi SV6 | |
| 30G Needles | Braun | 4656300 | 30G x ½” |
| 1ml syringes | Braun | 9161406V | Tuberculin syringe |
| Disposable scalpel | Feather | 2975#15 | #15 blade |
| Standard surgical scissors | Fine Science Tools | 14001-13 | Sharp/blunt, for opening skin and muscle |
| Adson forcep | Fine Science Tools | 1101-12 | With teeth, for grasping skin and muscle |
| Alm retractor | Fine Science Tools | 17008-07 | With blunt teeth, for spreading abdominal cavity open during surgery |
| Gauze swabs | Lohmann & Rauscher | 18504 | 7.5 x 7.5 cm, should be autoclaved prior to use |
| Cotton-tipped applicator swabs | Lohmann & Rauscher | 11970 | Sterile |
| Fine-tipped foreceps | Fine Science Tools | 11063-07 | 0.4mm, angled tip, for dissecting hepatic artery |
| Vannas spring scissors | Fine Science Tools | 91500-09 | For delicate cutting |
| Micro-needle holder | Fine Science Tools | 12076-12 | For ligating gastroduodenal artery |
| Needle holder | Fine Science Tools | 12005-15 | Tungsten carbide jaws |
| 7-0 Prolene sutures | Ethicon | 8648H | Polypropylene suture with curved needle, for ligating gastroduodenal artery |
| 4-0 Vicryl sutures | Ethicon | V3040H | With curved needle attached |
| Infrared warming lamp | Beurer | IL11 | For maintaining body temperature post-operatively |
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