Name: transarterial administration of oncolytic viruses for locoregional therapy of orthotopic hcc in rats
Uploaded: 2016-04-15T13:00:00
Description: Watch this Scientific Journal Video about Transarterial Administration of Oncolytic Viruses for Locoregional Therapy of Orthotopic HCC in Rats at JoVE.com

This work is supported by the SFB 824 subprojects C6 and C7 (DFG Sonderforschungsbereich 824), German Research Foundation, Bonn, Germany.

Merk: Følgende trinn er utført i samsvar med retningslinjene i vår institusjon og de lokale myndighetene i Bayern, Tyskland. Alle forsøk på å reprodusere denne protokollen må gjøres i tilslutning med lokale retningslinjer for human behandling av dyr, som diktert av lokale dyr omsorg og bruk komité. For eksempel, mange institutter krever at sterile hansker er slitt under gnager kirurgi. Videre arbeid med virus må utføres i henhold til lokale forskrifter, ta vare for personlig og miljøsikkerhet. Hensyn må tas...

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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 ...

Leverkreft (HCC) er den femte vanligste kreftsykdommen i verden, og den tredje største årsaken til kreft-relaterte dødsfall, noe som gjør det til et betydelig helseproblem 1,2. For pasienter som ikke er kvalifisert for tumor reseksjon, eller de venter på levertransplantasjon, lokoregionalt terapi involverer transarterial embolisering (TAE) eller transarterial chemoembolization (TACE) brukes som standard palliativ omsorg 3,4. Disse behandlingene utnytte unike med dobbel blodtilførsel i leveren hvor svulster er lei nesten utelukkende ved lever arteriell blodstrøm, mens rundt leveren mottar mesteparten av sin blodforsyning fra portvenen 5,6. På grunn av de ekstremt begrensede efficacies av etablerte behandlingsformer for HCC har onkolytiske virus dukket opp som lovende alternative behandlingsformer. JX-594, nylig omdøpt Pexa-Vec, er en tymidinkinase-slettet vaccinia vektor, bevæpnet med granulocytt-mMakrofag koloni-stimulerende faktor (GM-CSF), som har gjennomført klinisk fase II-studie for HCC 7. Mer nylig har en rekombinant vesikulær stomatitt virus vektor (VSV) uttrykker humant interferon beta inne i en fase jeg klinisk studie for sorafenib-ildfast HCC (NCT01628640). Som onkolytiske virus gå nærmere få godkjent for klinisk anvendelse for HCC pasienter, behovet for en effektiv administreringsrute målrette multifokal sykdom er tydelig. Mens systemisk tilførsel er stort sett ineffektive på grunn av ineffektiv svulst transduksjon, kan intratumorale anvendelser begrenser effektiviteten av terapien til den injiserte tumor, slik uninjectable mikroskopiske lesjoner som er mottakelige for sykdomsprogresjon. Vi har etablert en fremgangsmåte for å isolere de leverarterien hos rotter for å administrere onkolytisk virusbehandling i en lokoregional måte å målrette ortotopisk HCC. Vi har vist at denne tilførselsvei resulterer i trygge og effective transduksjon av multifokale HCC knuter, noe som resulterer i betydelig overlevelse forlengelse i immun kompetente rotter 8-10. Her beskriver vi en fremgangsmåte for å få tilgang til, dissekere, og injisering i den hepatiske arterien hos rotter. En ordning av fremgangsmåten er vist i figur 1 (tidligere utgitt 9)

<table border="0" cellpadding="0" cellspacing="0" width="597">
	<tbody>
		<tr height="64">
			<td height="64" style="height:64px;width:144px;">Veterinary clippers</td>
			<td style="width:157px;">Aesculap</td>
			<td style="width:88px;">GT415</td>
			<td style="width:208px;">Small, cordless trimmer ideal for removing fur from surgical area</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:144px;">Stereomicroscope</td>
			<td style="width:157px;">Zeiss</td>
			<td style="width:88px;">Stemi SV6</td>
			<td style="width:208px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:144px;">30G Needles</td>
			<td style="width:157px;">Braun</td>
			<td align="right" style="width:88px;">4656300</td>
			<td style="width:208px;">30G x &#189;&#8221;</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:144px;">1ml syringes</td>
			<td style="width:157px;">Braun</td>
			<td style="width:88px;">9161406V</td>
			<td style="width:208px;">Tuberculin syringe</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:144px;">Disposable scalpel</td>
			<td style="width:157px;">Feather</td>
			<td style="width:88px;">2975#15</td>
			<td style="width:208px;">#15 blade&#160;</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:144px;">Standard surgical scissors</td>
			<td style="width:157px;">Fine Science Tools</td>
			<td style="width:88px;">14001-13&#160;</td>
			<td style="width:208px;">Sharp/blunt, for opening skin and muscle</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:144px;">Adson forcep</td>
			<td style="width:157px;">Fine Science Tools</td>
			<td style="width:88px;">1101-12</td>
			<td style="width:208px;">With teeth, for grasping skin and muscle</td>
		</tr>
		<tr height="64">
			<td height="64" style="height:64px;width:144px;">Alm retractor</td>
			<td style="width:157px;">Fine Science Tools</td>
			<td style="width:88px;">17008-07</td>
			<td style="width:208px;">With blunt teeth, for spreading abdominal cavity open during surgery</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:144px;">Gauze swabs</td>
			<td style="width:157px;">Lohmann &#38; Rauscher</td>
			<td align="right" style="width:88px;">18504</td>
			<td style="width:208px;">7.5 x 7.5 cm, should be autoclaved prior to use</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:144px;">Cotton-tipped applicator swabs</td>
			<td style="width:157px;">Lohmann &#38; Rauscher</td>
			<td align="right" style="width:88px;">11970</td>
			<td style="width:208px;">Sterile</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:144px;">Fine-tipped foreceps</td>
			<td style="width:157px;">Fine Science Tools</td>
			<td style="width:88px;">11063-07</td>
			<td style="width:208px;">0.4mm, angled tip, for dissecting hepatic artery</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:144px;">Vannas spring scissors</td>
			<td style="width:157px;">Fine Science Tools</td>
			<td style="width:88px;">91500-09</td>
			<td style="width:208px;">For delicate cutting</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:144px;">Micro-needle holder</td>
			<td style="width:157px;">Fine Science Tools</td>
			<td style="width:88px;">12076-12</td>
			<td style="width:208px;">For ligating gastroduodenal artery</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:144px;">Needle holder</td>
			<td style="width:157px;">Fine Science Tools</td>
			<td style="width:88px;">12005-15</td>
			<td style="width:208px;">Tungsten carbide jaws</td>
		</tr>
		<tr height="64">
			<td height="64" style="height:64px;width:144px;">7-0 Prolene sutures</td>
			<td style="width:157px;">Ethicon</td>
			<td style="width:88px;">8648H</td>
			<td style="width:208px;">Polypropylene suture with curved needle, for ligating gastroduodenal artery</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:144px;">4-0 Vicryl sutures</td>
			<td style="width:157px;">Ethicon</td>
			<td style="width:88px;">V3040H</td>
			<td style="width:208px;">With curved needle attached</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:144px;">Infrared warming lamp</td>
			<td style="width:157px;">Beurer</td>
			<td style="width:88px;">IL11</td>
			<td style="width:208px;">For maintaining body temperature post-operatively</td>
		</tr>
	</tbody>
</table>

transarterial administration of oncolytic viruses for locoregional therapy of orthotopic hcc in rats

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.

Med erfaring, kan en nesten 100% suksessrate (som betyr at arterien ble vellykket dissekert og injisert, og dyret overlevde operasjonen) oppnås. Imidlertid vil helsetilstanden rotte før kirurgi (grad av tumorbyrden, underliggende leverfunksjon, etc.) åpenbart spille en rolle i utfallet av kirurgiske inngrep. Etter kirurgi, kan rotter forventes å oppleve forbigående vekttap og en svak, forbigående økning av leverenzymer, selv om buffer alene, uten virus, ble injisert. ...

Watch this Scientific Journal Video about Transarterial Administration of Oncolytic Viruses for Locoregional Therapy of Orthotopic HCC in Rats at JoVE.com

Transarterial Administration of Oncolytic Viruses for Locoregional Therapy of Orthotopic HCC in Rats

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.

Transarterial Administrasjon av Onkolytiske virus for Lokal regional terapi av ortotopiske HCC i rotter

Hepatocellular carcinoma (HCC) is a disease with limited treatment options and poor prognosis.  In recent years, oncolytic virotherapies have proven ...

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