# Genes Tumor Vector Mode Results Authors/year/references Cytokines Canine 1 hIL-2 MEL Irradiated hIL-2 producing xenogeneic cells made by plasmid transfection Ex vivo/ p.t./ +SX + RX( = 16) NR, 480 d. Control: 1/treated: 6. Median survival: treated: 270 d/control: 75 d Quintin-Colonna et al., 1996 [2 ] 2 cIL-2 + enterotoxin-A STS Plasmid lipofection In vivo /i.t. + SX( = 16) 3 CR, 1 PR, 1 SD (84 d). Responders survival: >540 d w/o relapse. Thamm et al., 2003 [33 ] 3 cIL-2 OSA lung metastases Plasmid lipofection In vivo/ i.v./ +SX + CHT( = 20) 84 d: 1 CR, 2 PR, 4 SD. Median survival (s IV): treated: 82 d/Hist. Surg. controls: 58 d Dow et al., 2005 [36 ] 4 hGM-CSF MEL FSA OSA HGS Irradiated hGM-CSF producing autologous tumor cells plasmid transferred by gene gun Ex vivo/ i.d./ NATMEL ( = 10) 1 CR (300 d), 1 PR (300 d), 1 NED (250 d), 1 SD (125 d) FSA ( = 3) 1 PR, 1 NED (175 d) Hogge et al., 1998 [13 ] 5 cGM-CSF + enterotoxin-B MEL Plasmid lipofection In vivo/ i.t., p.t., i.l.n./NAT( = 12) median survival (stage III): treated: 168 d/Hist. Surgery controls: 105 d Dow et al., 1998 [14 ] 6 hGM-CSF LYP Plasmid lipofection In vivo/ i.d./ +CHTVaccine ( = 26)/placebo ( = 26). Median survival 474 d/342 d Turek et al., 2007 [40 ] 7 hIL-2 + hGM-CSF HSV-tk MEL Plasmid lipofection: HSV-tk + GCV. Irradiated xenogeneic hIL-2 and hGM-CSF producing cells made by plasmid lipofection In vivo (SG)/i.t., Ex vivo (CPXC)/i.t./NATCombined treatment ( = 45)/surgery controls ( = 23). Median survival: 160 d/82 d. Metastasis-free median survival: >509 d/133 d. Combined treatment: CR = 16%, PR = 31%, SD = 27%, PD = 27%. Finocchiaro et al., 2008 [28 ] 8 hIL-2 + hGM-CSF HSV-tk MEL Plasmid lipofection: HSV-tk + GCV. Irradiated xenogeneic hIL-2 and hGM-CSF producing cells made by plasmid lipofection In vivo (SG) − i.t./Ex vivo (CPXC) + (TV) − s.c./+SXCombined treatment ( = 100)/surgery controls ( = 51): local disease-free patients: 58%/6%. Metastasis-free patients: 78%/43%. Median survival: >1312 d/78 d. Metastasis-free median survival: >1312 d/112 d. Finocchiaro and Glikin, 2008 [29 ] 9 hIL-2 + hGM-CSF HSV-tk MEL Plasmid lipofection: HSV-tk + GCV. Irradiated xenogeneic hIL-2 and hGM-CSF producing cells made by plasmid lipofection In vivo (SG) − i.t. Ex vivo (CPXC) + (TV) − s.c./+SXCombined treatment ( = 283)/surgery controls ( = 135). Complete surgery arms: local disease-free patients: 81%/13%. Metastasis-free patients: 84%/32%. Median survival: >2848 d/99 d. Metastasis-free median survival: >2848 d/99 d. Finocchiaro and Glikin, 2012 [30 ] 10 hIL-2 + hGM-CSF cIFN-β + HSV-tk MEL Plasmid lipofection: HSV-tk + IFN-β + GCV. Plasmid lipofection: hIL-2 and hGM-CSF In vivo (SG) + (IFN-β ) − i.t. In vivo (hIL-2 + hGM-CSF) + (TV) − s.c./+SXCombined treatment ( = 301)/surgery controls ( = 162). Complete surgery arms: local disease-free patients: 83%/11%. Metastasis-free patients: 89%/44%. Median survival: >2211 d/109 d. Metastasis-free median survival: >2211 d/134 d. Finocchiaro et al., Submitted [31 ] Genes Cytokines Tumor Canine 11 hIL-2 + hGM-CSF cIFN-β + HSV-tk STS Plasmid lipofection: HSV-tk + IFN-β + GCV. Irradiated xenogeneic hIL-2 and hGM-CSF producing cells made by plasmid lipofection In vivo (SG) + (IFN-β ) − i.t. Ex vivo (CPXC) + (TV) − s.c/+SX.( = 11) 5 survived >2 years, 4 survived >1 year Finocchiaro et al., 2011 [34 ] 12 hIL-2 + hGM-CSF cIFN-β + HSV-tk OSA Plasmid lipofection: HSV-tk + IFN-β + GCV. Irradiated xenogeneic hIL-2 and hGM-CSF producing cells made by plasmid lipofection In vivo (SG) + (IFN-β ) − i.t. Ex vivo (CPXC) + (TV) − s.c/+SX.( = 6) 2 survived > 1 year Finocchiaro et al., 2012 [37 ] 13 hIFN-γ ASC Adenovirus In vivo − p.t. + (TV) − s.c./ +SX.( = 1) feasible. Disease-free survival >450 d Pluhar et al., 2010 [46 ] 14 fIL-12 + bleomycin SCC AMB MEL STS Naked plasmid + bleomycin electrotransfer In vivo − i.t./+CHT( = 5) 2 SSC and AMB resolved (56, 27, 9 months tumor-free). Reed et al., 2010 [45 ] 15 hIL-12 MCT PHS OSA MAC Naked plasmid electrotransfer In vivo − i.m/ ±SX ± CHT( = 6) CR: 2 of 3 MCT. SD: 1 PHS and 1 OSA. Cemazar et al., 2011 [47 ] 16 hIL-12 MCT Naked plasmid electrotransfer In vivo − i.t./ ±SX ± CHT( = 8) 50% median reduction of tumor volumes Pavlin et al., 2011 [48 ] Antigens Canine 17 hTyr MEL oral Naked plasmid − Jet injection In vivo − i.m./ ±SX ± RX( = 9) 1 CR, 2 NED, 1 SD (14 d)/median survival: 389 d. Historical controls (s II 150 d, s III 60 d). Humoral antibodies: 3/9. Bergman et al., 2003 [16 ] Liao et al., 2006 [17 ] 18 hTyr/mTyr mgp75/ hGM-CSF MEL Naked plasmid − Jet injection In vivo − i.m./ ±SX ± RXDifferent combinations and doses. Median survival stages II-III ( = 33) w/locoregional control: 569 d. 25/33 still alive at submission. Bergman et al., 2006 [20 ] 19 hTyr MEL oral Naked plasmid − Jet injection In vivo − i.m./ +SX ± RX( = 58) death attributable to MM ( = 15), to other causes ( = 16). Median survival: >750 d. Historical controls ( = 53): 324 d. Grosenbaugh et al., 2011 [18 ] 20 mTyr MEL digital Naked plasmid − Jet injection In vivo − i.m./ +SX + RX( = 58) median survival: >476 d. Surgery only historical controls: 365 d Manley et al., 2011 [21 ] Antigens Canine 21 hTyr MEL Naked plasmid − Jet injection In vivo − i.m/ +SX ± RX( = 22) no differences in PFS, DFI, or median survival with surgery controls ( = 23) Ottnod et al., 2013 [22 ] 22 hgp100 MEL Autologous adenovirus transduced dendritic cells Ex vivo − s.c./ +SX + RX( = 3) survival ranging 210–1440 d Gyorffy et al., 2005 [23 ] 23 hgp100 MEL Irradiated allogeneic hgp100 expressing tumor cells plasmid transferred by gene gun Ex vivo − i.d./ NAT( = 34) median survival: responders (CR, PR, SD) ( = 12), 337 d/nonresponders ( = 20), 95 d Alexander et al., 2006 [24 ] 24 cTERT cTERT-LTB LYP Naked plasmid electroporation + adenovirus 6 In vivo − i.m./ +CHT( = 14) GT + CHT /( = 8) CHT . Median survival: >96 wks/28 wks. Time to first relapse: >26 wks/12 wks. Peruzzi et al., 2010 [42 ] 25 cTERT LYP Electroporation + Adenovirus In vivo − i.m./ +CHT( = 21) VAC/( = 21) CTR median survival: VAC 76.1 w/CTR 29.3 w Gavazza et al., 2013 [43 ] 26 Emm55 LYP Plasmid/electroporation Ex vivo − i.v./ ±CHT( = 7) 1 CR > 555 d, 6 PR, 3 extended survival >2X Lawman et al., 2008 [41 ] 27 CSPG4 MEL Plasmid/electroporation In vivo − i.m./. +SX.( = 14) VAC/( = 13) CTR median survival: VAC 76.1 w/CTR 29.3 w Riccardo et al., 2014 [27 ] 28 cNHL-mRNA LYP Combination chemotherapy + NHL-mRNA loaded electroporation of CD40-activated B cells. Ex vivo RNA transfer/ +CHT( = 19) significant increase of lymphoma-specific survival following salvage therapy Sorenmo et al., 2011 [44 ] Specific ligands Canine 29 hCD40L MEL Adenovirus In vivo − i.t./. ±SX.( = 2) feasible and manageable toxicity 1 NED 401 d 1 PR >150 d von Euler et al., 2008 [25 ] 30 hCD40L MEL Adenovirus In vivo − i.t./. ±SX.( = 19) median survival: responders (CR, PR, SD) ( = 17), 160 d/nonresponders ( = 2) Westberg et al., 2013 [26 ] 31 hFasL MEL Plasmid lipofection In vivo − i.t./ ±SX ± RX( = 5) GT: 3 TR, 1 SD/CT: 2 CR, 2 PR Survival: 168–574 d Bianco et al., 2003 [15 ] 32 hFasL OSA Adenovirus In vivo − i.t./ +SX( = 48) Ad-FasL-mediated tumor inflammation improved median survival (score 2/3: 359 d; controls: 221 d) Modiano et al., 2012 [38 ]