Administración transarterial de virus oncolíticos para locorregional Terapia de HCC ortotópico en ratas
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
El carcinoma hepatocelular (CHC) es el quinto cáncer más frecuente en todo el mundo, y la tercera causa principal de muerte relacionada con el cáncer, por lo que es un importante problema de salud 1,2. Para los pacientes que no son candidatos para la resección del tumor, o aquellos que esperan un trasplante de hígado, la terapia locorregional la técnica de embolización transarterial (TAE) o la quimioembolización transarterial (TACE) se aplican como estándar de cuidados paliativos 3,4. Estas terapias se aprovechan de la característica única de doble suministro de sangre en el hígado mediante el cual los tumores se alimentan casi exclusivamente por el flujo sanguíneo arterial hepática, mientras que el hígado circundante recibe la mayor parte de su suministro de sangre de la vena porta 5,6.
Debido a las eficacias extremadamente limitadas de las terapias establecidas para el CHC, virus oncolíticos han surgido como alternativas terapéuticas prometedoras. JX-594, recientemente rebautizada Pexa-Vec, es una quinasa eliminado en vectores vaccinia timidina, armado de granulocitos-macrophage factor estimulante de colonias (GM-CSF), que se ha completado la fase II de ensayos clínicos para el CHC 7. Más recientemente, un vector recombinante virus de la estomatitis vesicular (VSV) que expresa interferón beta humano ha entrado en una fase I de ensayos clínicos para el CHC sorafenib-refractario (NCT01628640). Como los virus oncolíticos se acercan más a la obtención de la aprobación para la aplicación clínica de HCC en pacientes, la necesidad de una vía de administración eficaz para apuntar enfermedad multifocal es evidente. Mientras que la administración sistémica es en gran parte ineficaz debido a la transducción de tumor ineficiente, aplicaciones intratumorales podrían limitar la eficacia de la terapia para el tumor inyectado, dejando lesiones microscópicas uninjectable susceptibles a la progresión de la enfermedad.
Hemos establecido un método de aislamiento de la arteria hepática en ratas para administrar la terapia virus oncolítico de una manera locorregional para apuntar HCC ortotópico. Hemos demostrado que esta vía de administración da como resultado seguro y effective la transducción de nódulos HCC multifocales, lo que resulta en una prolongación significativa la supervivencia en ratas inmunocompetentes 8-10. A continuación, se describe el método de acceso, disección, e inyectar en la arteria hepática en ratas. Un esquema del procedimiento se muestra en la Figura 1 (previamente publicado 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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