ラットにおける同所性肝細胞癌の局所領域治療のための腫瘍溶解性ウイルスのTransarterial管理
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
肝細胞癌（HCC）は重大な健康問題1,2作り、世界中で5番目に普及している癌、および癌関連死因の第3位です。腫瘍切除の対象とならない患者、または肝移植を待っている人のために、動脈塞栓術（TAE）または肝動脈化学塞栓療法（TACE）を含む局所領域療法は、標準的な緩和ケア3,4として適用されます。周囲の肝臓は門脈5,6からの血液供給の大部分を受け取りながら、これらの治療法は、腫瘍は肝動脈血流によってほぼ独占的に供給され、それによって肝臓でのデュアル血液供給のユニークな機能を活用します。
HCCの確立された治療法の非常に限られた有効性のために、腫瘍溶解性ウイルスは、有望な代替治療薬として浮上しています。 JX-594、最近PEXA-VECの名前を変更し、顆粒球メートルで武装し、チミジンキナーゼ欠損ワクシニアベクターでありますHCC 7のための第II相臨床試験を完了したacrophageコロニー刺激因子（GM-CSF）、。さらに最近では、ヒトインターフェロンβを発現する組換え水疱性口内炎ウイルスベクター（VSV）は、ソラフェニブ難治性HCC（NCT01628640）のための第I相臨床試験に入りました。腫瘍溶解性ウイルスは患者におけるHCCの臨床応用のための承認を得るに近づくように、多焦点疾患を標的とするための効果的な投与経路の必要性は明らかです。全身送達が非効率的な腫瘍伝達にほとんど効果があるが、腫瘍内のアプリケーションは、疾患の進行の影響を受けやすいuninjectable顕微鏡的病変を残して、注入された腫瘍に対する治療の有効性を制限することができます。
私たちは、同所性HCCをターゲットとする局所領域の方法で腫瘍溶解性ウイルス療法を管理するために、ラットにおける肝動脈を単離する方法を確立しました。我々は実証されていること、安全かつeffectiでこの投与経路結果免疫適格ラット8-10で有意な生存の延長で、その結果、多焦点HCC結節の伝達をVEの。ここでは、アクセス解剖、およびラットにおける肝動脈内に注入する方法について説明します。手順のスキームを図1に示されている（以前は9日公開）
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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