Transarterial Administração de Oncolíticos Vírus para loco-regional Terapia de Ortotópico HCC em Ratos
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
O carcinoma hepatocelular (HCC) é o quinto câncer mais comum no mundo, ea terceira principal causa de morte relacionada ao câncer, tornando-se um problema de saúde significativo 1,2. Para os pacientes que não são elegíveis para ressecção do tumor, ou aqueles aguardando transplante hepático, terapia loco-regional envolvendo embolização transarterial (TAE) ou quimioembolização transarterial (TACE) são aplicados como cuidados paliativos padrão 3,4. Estas terapias explorar a característica única de fornecimento de sangue duplo, no qual os tumores do fígado são alimentados quase exclusivamente pelo fluxo de sangue arterial hepático, enquanto o fígado circundante recebe a maioria do seu fornecimento de sangue a partir da veia portal de 5,6.
Devido às eficácias muito limitado de terapias estabelecidas de carcinoma hepatocelular, os vírus oncolí ticos têm emergido como agentes terapêuticos alternativos promissores. JX-594, recentemente renomeado Pexa-Vec, é um excluído-quinase vector vaccinia timidina, armado com granulócitos-macrophage-fator estimulador de colônias (GM-CSF), que foi concluída a fase II de ensaios clínicos para HCC 7. Mais recentemente, um vector recombinante do vírus da estomatite vesicular (VSV) que expressam o interferão-beta humano entrou numa fase I de ensaios clínicos para HCC-sorafenib refractário (NCT01628640). Como os vírus oncolí ticos aproximar-se a obtenção de aprovação para a aplicação clínica de HCC em pacientes, a necessidade de uma via de administração eficaz para alvejar doença multifocal é evidente. Enquanto a libertação sistémica é largamente ineficaz devido a transdução do tumor ineficiente, aplicações intratumorais poderia limitar a eficácia da terapia para o tumor injectado, deixando lesões microscópicas uninjectable sensíveis à progressão da doença.
Nós estabelecemos um método de isolamento da artéria hepática em ratos para administrar oncolytic vírus terapia de forma loco-regional para alvejar HCC ortotópico. Nós demonstramos que esta via de administração resultados em segurança e effective transdução de nódulos HCC multifocais, resultando no prolongamento de sobrevivência significativo em ratos imuno-competentes 8-10. Aqui, descrevemos o método de acesso, dissecando, e injetar na artéria hepática em ratos. Um esquema do processo é mostrado na Figura 1 (publicado anteriormente 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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