Transarterial Administration av cancerläkemedel Virus för Lokalregionala behandlingar av Orthotopic HCC hos råttor
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
Hepatocellulär cancer (HCC) är den femte vanligaste cancer i världen, och den tredje ledande orsaken till cancerrelaterad död, vilket gör det en betydande hälsoproblem 1,2. För patienter som inte är berättigade till tumörresektion, eller som väntar på levertransplantation, lokoregional terapi involverar transarterial embolisering (TAE) eller transarterial chemoembolization (TACE) tillämpas som standard palliativ vård 3,4. Dessa behandlingar utnyttja den unika funktionen av dubbel blodtillförsel i levern, varigenom tumörer matas nästan uteslutande genom lever arteriellt blodflöde, medan den omgivande lever får huvuddelen av sin blodtillförsel från portalvenen 5,6.
På grund av de extremt begränsade effektivitet etablerade terapier för HCC har onkolytiska virus dykt upp som lovande alternativa läkemedel. JX-594, nyligen omdöpt Pexa-Vec, är en tymidinkinas-raderade vacciniavektorn, beväpnade med granulocyt-macrophage kolonistimulerande faktor (GM-CSF), som har slutfört fas II-studie för HCC 7. På senare tid har en rekombinant vesikulär stomatit virusvektor (VSV) som uttrycker humant interferon-beta in i en fas I klinisk studie för sorafenib-refraktär HCC (NCT01628640). Som onkolytiska virus närma sig få godkännande för klinisk tillämpning för HCC patienter, behovet av en effektiv administreringssätt rikta multifokal sjukdom är uppenbar. Även systemisk leverans är i hög grad ineffektivt på grund av ineffektiv tumöromvandling kan intratumorala applikationer begränsa effekten av terapin till det injicerade tumören, vilket uninjectable mikroskopiska skador som är mottagliga för sjukdomsprogression.
Vi har etablerat ett förfarande för isolering leverartären hos råttor för att administrera onkolytiskt virus terapi i en lokoregional sätt att rikta orthotopic HCC. Vi har visat att detta administreringssätt resulterar i säker och effective transduktion av multifokala HCC knutor, vilket resulterar i signifikant överlevnads förlängning i immunkompetenta råttor 8-10. Här beskriver vi hur man får tillgång, dissekera, och injektion i leverartären hos råttor. Ett system av förfarandet visas i figur 1 (tidigare publicerats 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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