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. 2014:1114:413-26.
doi: 10.1007/978-1-62703-761-7_27.

Portal vein delivery of viral vectors for gene therapy for hemophilia

Alexandra Sherman  1 , Alexander Schlachterman, Mario Cooper, Elizabeth P Merricks, Robin A Raymer, Dwight A Bellinger, Roland W Herzog, Timothy C Nichols
Affiliations

Portal vein delivery of viral vectors for gene therapy for hemophilia

Alexandra Sherman et al. Methods Mol Biol. 2014.

Abstract

The liver is a very complex organ with a large variety of functions, making it an attractive organ for gene replacement therapy. Many genetic disorders can be corrected by delivering gene products directly into the liver using viral vectors. In this chapter, we will describe gene delivery via portal vein administration in mice and dogs to correct the blood coagulation disorder hemophilia B. Although there are multiple delivery routes for both viral and non-viral vectors in animals, portal vein administration delivers vectors directly and efficiently into the liver. Complete correction of murine hemophilia B and multi-year near-correction of canine hemophilia B have been achieved following portal vein delivery of adeno-associated viral (AAV) vectors expressing factor IX from hepatocyte-specific promoters. Peripheral vein injection can lead to increased vector dissemination to off-target organ such as the lung and spleen. Below, we will describe portal vein injection delivery route via laparotomy.

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Figures

Fig. 1
Fig. 1
Portal vein delivery of gene therapy vector to a mouse. (a) Displacement of intestinal tract and pancreas to gain access to portal vein. (b) Exposure of portal vein. The area directly caudal to the portal vein must be flat enough to allow a needle to enter the vessel without inhibiting the syringe. A Q-tip is used to “retract” the pancreas to put tension on the vessel. (c) Insertion of needle through the multiple layers to enter the portal vein. One should see the bevel of the needle in the vessel. (d) Vector administration. Blanching of the liver should be obvious when depressing the plunger while observing the liver
Fig. 2
Fig. 2
Portal vein delivery of gene therapy vector to a hemophilic dog. Panel 1: The spleen is exteriorized via a midline laparotomy incision. Panel 2: A 3–5 French balloon-tipped catheter is prepared by inflating the balloon to check for leaks. Panel 3: A small splenic vein is isolated and a venotomy is performed. Panel 4: The balloon-tipped catheter is inserted in the venotomy site and passed antegrade to the hepatic portal vein, usually 10–20 cm. Panel 5: With the balloon inflated, the gene therapy vector is administered to the liver. Panel 6: The spleen is returned to the abdomen. Panels 7 and 8: The laparotomy incision is repaired with careful attention to bleeding vessels
See this image and copyright information in PMC

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