Original Communications| Volume 129, ISSUE 3, P324-334, March 2001

Specific organ gene transfer in vivo by regional organ perfusion with herpes viral amplicon vectors: Implications for local gene therapy


      Background. Many gene therapy strategies would benefit from efficient, regional organ delivery of therapeutic genes. Methods. Regional perfusions of lung, liver, or bladder were performed to determine if rapid and efficient gene transfer can be accomplished in vivo, and to determine if in vivo gene transfer can be limited to the organ of interest. In addition, herpes simplex virus tumor necrosis factor (HSVtnf), carrying the human tumor necrosis factorα gene was used as a treatment for methylcholanthrene sarcoma in a syngeneic lung metastases model in Fisher rats. Results. A 20-minute perfusion using HSV carrying β-galactosidase (HSVlac) produced significant expression of this marker gene isolated to the target organs, without organ-specific tissue injury or inflammation. Regional perfusion of organs with HSV carrying the cytokine gene tumor necrosis factor α also resulted in high-level local organ production of this cytokine (2851 ± 53 pg/g tissue in perfused lung versus 0 for the contralateral lung). For the current vector construct, expression of the gene of interest peaked between 2 and 4 days and was undetectable by 2 weeks after perfusion. In animals undergoing perfusion as treatment for pulmonary sarcoma, there was no difference between tumor counts in lungs perfused with HSVlac (17 ± 6) or HSVtnf (22 ± 8), but either treatment resulted in lower tumor counts than controls (111 ± 24 nodules per lung, P <.02). Conclusions. Regional organ perfusion using herpes viral vectors is an effective and well-tolerated in vivo method of transiently delivering potentially toxic gene products to target organs in directing gene therapy. Regional lung perfusion with HSV amplicons reduces tumor burden in a rat model of pulmonary metastases, though HSVtnf cannot be demonstrated to augment the cytopathic effect of the HSV amplicon alone in the current model. (Surgery 2001;129:324-34.)
      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to Surgery
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Lienard D
        • Ewalenko P
        • Delmotte JJ
        • Renard N
        • Lejeune FJ.
        High-dose recombinant tumor necrosis factor alpha in combination with interferon gamma and melphalan in isolation perfusion of the limbs for melanoma and sarcoma.
        J Clin Oncol. 1992; 10: 52-60
        • Mack CA
        • Magovern CJ
        • Budenbender KT
        • Patel SR
        • Schwarz EA
        • Zanzonico P
        • et al.
        Salvage angiogenesis induced by adenovirus-mediated gene transfer of vascular endothelial growth factor protects against ischemic vascular occlusion.
        J Vasc Surg. 1998; 27: 699-709
        • Folkman J.
        Angiogenesis in cancer, vascular, rheumatoid and other disease. A review.
        Nat Med. 1995; 1: 27-31
        • Fong Y
        • Federoff HJ
        • Brownlee M
        • Blumberg D
        • Blumgart LH
        • Brennan MF.
        Rapid and efficient gene transfer in human hepatocytes by herpes viral vectors.
        Hepatology. 1995; 22: 723-729
        • Hudde T
        • Rayner SA
        • de Alwis M
        • Thrasher AJ
        • Smith J
        • Coffin RS
        • et al.
        Adeno-associated and herpes simplex viruses as vectors for gene transfer to the corneal endothelium.
        Cornea. 2000; 19: 369-373
        • Akkaraju GR
        • Huard J
        • Hoffman EP
        • Goins WF
        • Pruchnic R
        • Watkins SC
        • et al.
        Herpes simplex virus vector-mediated dystrophin gene transfer and expression in MDX mouse skeletal muscle.
        J Gene Med. 1999; 1: 280-289
        • Goya RG
        • Rowe J
        • Sosa YE
        • Tomasec P
        • Lowenstein PR
        • Castro MG.
        Use of recombinant herpes simplex virus type 1 vectors for gene transfer into tumour and normal anterior pituitary cells.
        Mol Cell Endocrinol. 1998; 139: 199-207
        • Geller AI
        • Breakefield XO.
        A defective HSV-1 vector expresses Escherichia coli B-galactosidase in cultured peripheral neurons.
        Science. 1988; 241: 1667-1669
      1. Cell biology: a laboratory manual. Growth of replication defective herpes virus amplicon vectors and their use for gene transfer. CHS Press, Cold Spring Harbor (NY)1996
        • Geller AI
        • Keyomarsi K
        • Bryan J
        • Pardee AB.
        An efficient deletion mutant packaging system for defective herpes simplex virus vectors: potential applications to human gene therapy and neuronal physiology.
        Proc Natl Acad Sci U S A. 1990; 87: 8950-8954
        • Bergold P
        • Cassaccia-Bonnefil P
        • Xiu-Liu Z
        • Federoff H.
        Transsynaptic neuronal loss induced in hippocampal slice cultures by a herpes simplex virus vector expressing the GluR6 subunit of the kainate receptor.
        Proc Natl Acad Sci U S A. 1993; 90: 6165-6169
        • Linnik M
        • Zahos P
        • Geschwind M
        • Federoff H.
        Expression of bcl-2 from a defective herpes simplex virus-1 vector limits neuronal death in focal cerebral ischemia.
        Stroke. 1995; 26: 1670-1675
        • Xu H
        • Federoff H
        • Maragos J
        • Parada LF
        • Kessler J
        Viral transduction of trk A into cultured nodose and spinal motor neurons conveys NGF responsiveness.
        Dev Biol. 1994; 163: 152-161
        • Johnson P
        • Miyanohara A
        • Levine F
        • Cahill T
        • Friedmann T.
        Cytotoxicity of a replication-defective mutant of herpes simplex virus type I.
        J Virol. 1992; 66: 2952-2965
        • Paterson T
        • Everett RA.
        A prominent serine-rich region in Vmw175, the major transcriptional regulator protein of herpes simplex virus type 1, is not essential for virus growth in tissue culture.
        J Gen Virol. 1990; 71: 1775-1783
        • Geschwind M
        • Lu B
        • Federoff HJ.
        Expression of neurotrophic genes from herpes simplex virus type I vectors modifying neuronal phenotype.
        in: Providing pharmacological access to the brain: alternative approaches. Academic Press, New York1994: 462-482
        • Weksler B
        • Schneider A
        • Ng B
        • Burt M.
        Isolated single lung perfusion in the rat: an experimental model.
        J Appl Physiol. 1993; 74: 2736-2739
        • Dannenberg AM
        • Suga M
        Histochemical stains for macrophages in cell smears and tissue sections: β-galactosidase, acid phosphatase, nonspecific esterase, succinic dehydrogenase, and cytochrome oxidase.
        in: Methods for studying mononuclear phagocytes. Academic Press, New York1981: 282-284
        • Huggett AS
        • Nixon DA
        Use of glucose oxidase, peroxidase, and O-dianisidine in determination of blood and urinary glucose.
        Lancet. 1957; ii: 368-370
        • Jarnagin WR
        • Debs RJ
        • Wang SS
        • Bissell DM.
        Cationic lipid-mediated transfection of liver cells in primary culture.
        Nucleic Acids Res. 1992; 20: 4205-4211
        • Paquereau L
        • Le Cam A.
        Electroporation-mediated gene transfer into hepatocytes: preservation of a growth hormone response.
        Anal Biochem. 1992; 204: 147-151
        • Sun WH
        • Burkholder JK
        • Sun J
        • Culp J
        • Turner J
        • Lu XG
        • et al.
        In vivo cytokine gene transfer by gene gun reduces tumor growth in mice.
        Proc Natl Acad Sci U S A. 1995; 92: 2889-2893
        • Somasundaram C
        • Tournier I
        • Feldmann G
        • Bernuau D.
        Increased efficiency of gene transfection in primary cultures of adult rat hepatocytes stimulated to proliferate: a comparative study using the lipofection and the calcium phosphate precipitate methods.
        Cell Biol Int Rep. 1992; 16: 653-662
        • Jeng KS
        • Chiang HJ.
        Delayed formation of gallstone after transcatheter arterial embolization for hepatocellular carcinoma. Is elective cholecystectomy advisable during hepatectomy?.
        Arch Surg. 1989; 124: 1319-1322
        • Seifer M
        • Hohne M
        • Schaefer S
        • Gerlich WH.
        In vitro tumorigenicity of hepatitis B virus DNA and HBx protein.
        J Hepatol. 1991; 13: S61-S65
        • Salmons B
        • Gunzburg WH.
        Targeting of retroviral vectors for gene therapy.
        Hum Gene Ther. 1993; 4: 129
        • Cardoso JE
        • Branchereau S
        • Jeyarah PR
        • Houssin D
        • Danos O
        • Heard JM.
        In situ retrovirus-mediated gene transfer into dog liver.
        Hum Gene Ther. 1993; 4: 411-418
        • Moscioni AD
        • Rozga J
        • Neuzil DF
        • Overell RW
        • Holt JT
        • Demetriou AA.
        In vivo regional delivery of retrovirally mediated foreign genes to rat liver cells: need for partial hepatectomy for successful foreign gene expression.
        Surgery. 1993; 113: 304-311
        • Lattime EC
        • Lee SS
        • Eisenlohr LC
        • Mastrangelo MJ
        In situ cytokine gene transfection using vaccinia virus vectors. [Review] [98 refs].
        Sem Oncol. 1996; 23: 88-100
        • Rao JB
        • Chamberlain RS
        • Bronte V
        • Carroll MW
        • Irvine KR
        • Moss B
        • et al.
        IL-12 is an effective adjuvant to recombinant vaccinia virus- based tumor vaccines: enhancement by simultaneous B7-1 expression.
        J Immunol. 1996; 156: 3357-3365
        • Lee SS
        • Eisenlohr PA
        • McCue PA
        • Mastrangelo MJ
        • Lattime EC.
        Intravesical gene therapy: in vivo gene transfer using recombinant vaccinia virus vectors.
        Cancer Res. 1994; 54: 3325-3328
        • DeMatteo RP
        • Chu G
        • Ahn M
        • Chang E
        • Burke C
        • Raper SE
        • et al.
        Immunologic barriers to hepatic adenoviral gene therapy for transplantation.
        Transplantation. 1997; 63: 315-319
        • McElvaney NG
        • Crystal RG
        IL-6 release and airway administration of human CFR cDNA adenovirus vector [letter].
        Nat Med. 1995; 1: 182-184
        • Asher AL
        • Mule JJ
        • Kasid A
        • Restifo NP
        • Salo JC
        • Reichert CM
        • et al.
        Murine tumor cells transduced with the gene for tumor necrosis factor-alpha.
        J Immunol. 1991; 146: 3227-3234
        • Rosenberg SA
        • Anderson WF
        • Blaese M
        • Hwu P
        • Yannelli JR
        • Yang JC
        • et al.
        The development of gene therapy for the treatment of cancer.
        Ann Surg. 1993; 218: 455-463
        • Hwu P
        • Yannelli J
        • Kriegler M
        • Anderson WF
        • Perez C
        • Chiang Y
        • et al.
        Functional and molecular characterization of tumor-infiltrating lymphocytes transduced with tumor necrosis factor-alpha cDNA for the gene therapy of cancer in humans.
        J Immunol. 1993; 150: 4104-4115
        • Karpoff HM
        • D'Angelica M
        • Blair S
        • Brownlee MD
        • Federoff H
        • Fong Y
        Prevention of hepatic tumor metastases in rats with herpes viral vaccines and gamma-interferon.
        J Clin Invest. 1997; 99: 799-804
        • Tung C
        • Federoff HJ
        • Brownlee M
        • Karpoff H
        • Weigel T
        • Brennan MF
        • et al.
        Rapid production of IL-2 secreting tumor cells by HSV-mediated gene transfer: implications for autologous vaccine production.
        Hum Gene Ther. 1996; 7: 2217-2224
        • Fong Y
        • Federoff HJ
        • Brownlee M
        • Blumberg D
        • Blumgart LH
        • Brennan MF.
        Rapid and efficient gene transfer in human hepatocytes by herpes viral vectors.
        Hepatology. 1995; 22: 723-729
        • Lowy AM
        • Curley SA.
        Clinical and preclinical trials of isolated liver perfusion for advanced liver tumors.
        Surg Oncol Clin N Am. 1996; 5: 429-441
        • Remick DG
        • Kunkel SL
        Toxic effects of cytokines in vivo. [Review] [37 refs].
        Lab Invest. 1989; 60: 317-319
        • Fausto N
        • Mead JE.
        Role of protooncogenes and transforming growth factors in normal and neoplastic liver growth.
        Prog Liver Dis. 1990; 9: 57-71
        • Engelhardt JF
        • Litzky L
        • Wilson JM.
        Prolonged transgene expression in cotton rat lung with recombinant adenoviruses defective in E2a.
        Hum Gene Ther. 1994; 5: 1217-1229
        • Smythe WR
        • Hwang HC
        • Elshami AA
        • Amin KM
        • Albelda SM
        • Kaiser LR
        Differential sensitivity of thoracic malignant tumors to adenovirus-mediated drug sensitization gene therapy.
        J Thorac& Cardiovasc Surg. 1995; 109: 626-630
        • Boviatsis EJ
        • Chase M
        • Wei MX
        • Tamiya T
        • Hurford RK
        • Kowall NW
        • et al.
        Gene transfer into experimental brain tumors mediated by adenovirus, herpes simplex virus, and retrovirus vectors.
        Hum Gene Ther. 1994; 5: 183-191
        • D'Angelica M
        • Karpoff H
        • Halterman M
        • Ellis J
        • Klimstra D
        • Edelstein D
        • et al.
        In vivo interleukin-2 gene therapy of established tumors with herpes simplex amplicon vectors.
        Cancer Immunol Immunother. 1999; 47: 265-271
        • Hickman MA
        • Malone RW
        • Lehmann-Bruinsma K
        • Sih TR
        • Knoell D
        • Szoka FC
        • et al.
        Gene expression following direct injection of DNA into liver.
        Hum Gene Ther. 1994; 5: 1477-1483
        • Pakzaban P
        • Geller AI
        • Isacson O.
        Effect of exogenous nerve growth factor on neurotoxicity of and neuronal gene delivery by a herpes simplex amplicon vector in the rat brain.
        Hum Gene Ther. 1994; 5: 987-995
        • Chao J
        • Jin L
        • Chen LM
        • Chen VC
        • Chao L.
        Systemic and portal vein delivery of human kallikrein gene reduces blood pressure in hypertensive rats.
        Hum Gene Ther. 1996; 7: 901-911
        • Miyanohara A
        • Johnson PA
        • Elam RL
        • Dai Y
        • Witztum JL
        • Verma IM
        • et al.
        Direct gene transfer to the liver with herpes simplex virus type 1 vectors: transient production of physiologically relevant levels of circulating factor IX.
        New Biologist. 1992; 4: 238-246
        • Rainov NG
        • Zimmer C
        • Chase M
        • Kramm CM
        • Chiocca EA
        • Weissleder R
        • et al.
        Selective uptake of viral and monocrystalline particles delivered intra-arterially to experimental brain neoplasms.
        Hum Gene Ther. 1995; 6: 1543-1552
        • Rome JJ
        • Shayani V
        • Newman KD
        • Farrell S
        • Lee SW
        • Virmani R
        • et al.
        Adenoviral vector-mediated gene transfer into sheep arteries using a double-balloon catheter.
        Hum Gene Ther. 1994; 5: 1249-1258
        • Nabel EG
        • Yang Z
        • Muller D
        • Chang AE
        • Gao X
        • Huang L
        • et al.
        Safety and toxicity of catheter gene delivery to the pulmonary vasculature in a patient with metastatic melanoma.
        Hum Gene Ther. 1994; 5: 1089-1094
        • Glorioso JC
        • DeLuca NA
        • Fink DJ.
        Development and application of herpes simplex virus vectors for human gene therapy 1.
        Annu Rev Microbiol. 1995; 49: 675-710
        • Fraefel C
        • Song S
        • Lim F
        • Lang P
        • Yu L
        • Wang Y
        • et al.
        Helper virus-free transfer of herpes simplex virus type 1 plasmid vectors into neural cells 1.
        J Virol. 1996; 70: 7190-7197
        • Weksler B
        • Blumberg D
        • Lenert JT
        • Ng B
        • Fong Y
        • Burt ME.
        Isolated single-lung perfusion with TNF-alpha in a rat sarcoma lung metastases model.
        Ann Thorac Surg. 1994; 58: 328-332
        • Pechan PA
        • Fotaki M
        • Thompson RL
        • Dunn R
        • Chase M
        • Chiocca EA
        • et al.
        A novel ‘piggyback’ packaging system for herpes simplex virus amplicon vectors 1.
        Hum Gene Ther. 1996; 7: 2003-2013
        • Hodgson HJ.
        Primary hepatocellular carcinoma.
        in: 1st edition. Surgery of the liver and biliary tract. Churchill Livingston, London1988: 1129-1134
        • Boring CC
        • Squires TS
        • Tong T.
        Cancer statistics, 1993.
        CA Cancer J Clin. 1994; 43: 7-26
        • Gilbert HA
        • Kagan AR.
        Metastases: incidence, detection, and evaluation without histologic confirmation.
        in: Fundamental aspects of metastasis. North-Holland Publishing Company, Amsterdam1976: 385-405