Transarterial Administration af onkolytiske virus for Lokoregional Therapy af ortotopisk HCC i rotter
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
Hepatocellulært carcinom (HCC) er den femte mest udbredte kræft på verdensplan, og den tredje hyppigste årsag til kræft-relaterede dødsfald, gør det en væsentlig sundhedsproblem 1,2. For patienter, der ikke er berettiget til tumor resektion, eller dem afventer levertransplantation, locoregional terapi involverer transarterial embolisering (TAE) eller transarterial chemoembolization (TACE) anvendes som standard palliativ pleje 3,4. Disse behandlinger udnytte den unikke egenskab af dobbelt blodforsyning i leveren, hvor tumorer fodres næsten udelukkende af hepatisk arteriel blodgennemstrømning, mens den omgivende leveren modtager størstedelen af sin blodforsyning fra portalen vene 5,6.
På grund af de ekstremt begrænsede effektiviteter af etablerede terapier for HCC har oncolytiske vira dukket op som lovende alternative terapeutiske midler. JX-594, for nylig omdøbt Pexa-Vec, er et thymidin kinase-slettet vaccinia vektor, bevæbnet med granulocyt-macrophage koloni-stimulerende faktor (GM-CSF), som har afsluttet klinisk fase II studie med HCC 7. For nylig er et rekombinant vesikulær stomatitis-virus vektor (VSV), der udtrykker humant interferon-beta ind i en fase I klinisk studie med sorafenib-refraktær HCC (NCT01628640). Som onkolytiske virus komme tættere på at opnå godkendelse til klinisk ansøgning om HCC patienter, behovet for en effektiv administration rute målrette multifokal sygdom er tydelig. Mens systemisk afgivelse er stort set ineffektive som følge af ineffektiv tumor transduktion, kunne intratumorale applikationer begrænse effektiviteten af terapien til det injicerede tumor, efterlader uninjectable mikroskopiske læsioner modtagelige for sygdomsprogression.
Vi har etableret en fremgangsmåde til isolering af den hepatiske arterie i rotter at administrere onkolytiske virus terapi i en locoregional måde at målrette ortotopisk HCC. Vi har vist, at denne administration rute resulterer i sikker og effective transduktion af multifokale HCC knuder, hvilket resulterer i en betydelig overlevelse forlængelse i immunkompetente rotter 8-10. Her beskriver vi den måde at få tilgang, dissekere, og injektion i den hepatiske arterie i rotter. En plan for fremgangsmåden er vist i figur 1 (tidligere offentliggjorte 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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