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Doxycycline Inducible Melanogenic Vaccinia Virus as Theranostic Anti-Cancer Agent.

Kirscher L, Deán-Ben XL, Scadeng M, Zaremba A, Zhang Q, Kober C, Fehm TF, Razansky D, Ntziachristos V, Stritzker J, Szalay AA - Theranostics (2015)

Bottom Line: In this study melanin production was detected after exogenous addition of doxycycline in two different tumor xenograft mouse models.Furthermore, it was confirmed that this novel vaccinia virus strain still facilitated signal enhancement as detected by MRI and optoacoustic tomography.At the same time we demonstrated an enhanced oncolytic potential compared to the constitutively melanin synthesizing rVACV system.

View Article: PubMed Central - PubMed

Affiliation: 1. University of Würzburg, Department of Biochemistry, Am Hubland, 97074 Würzburg, Germany.

ABSTRACT
We reported earlier the diagnostic potential of a melanogenic vaccinia virus based system in magnetic resonance (MRI) and optoacoustic deep tissue imaging (MSOT). Since melanin overproduction lead to attenuated virus replication, we constructed a novel recombinant vaccinia virus strain (rVACV), GLV-1h462, which expressed the key enzyme of melanogenesis (tyrosinase) under the control of an inducible promoter-system. In this study melanin production was detected after exogenous addition of doxycycline in two different tumor xenograft mouse models. Furthermore, it was confirmed that this novel vaccinia virus strain still facilitated signal enhancement as detected by MRI and optoacoustic tomography. At the same time we demonstrated an enhanced oncolytic potential compared to the constitutively melanin synthesizing rVACV system.

No MeSH data available.


Related in: MedlinePlus

Oncolytic activity of inducible melanogenic rVACV strains. Photography A) and quantitative evaluation B) of plaque sizes in A594 cancer cells infected with melanogenic rVACV incubated in presence (+) or absence (-) of 1 µg/ml dox. Significantly larger mean maximum plaque diameter of non-induced not melanin producing GLV-1h462 infected A549 cancer cells compared to induced melanogenic GLV-1h462 infected A549 cells or GLV-1h460 infected A549 cancer cells (p < 0.001; n ≥ 50). MTT-Assay of A549 C) and PANC-1 D) cancer cells infected (MOI 0.01 or MOI 0.1) with the constitutively melanin synthesizing GLV-1h460, the non-melanogenic GLV-1h68 and GLV-1h312 and the dox inducible melanogenic GLV-1h462. The control rVACV showed no significant difference in oncolysis, regardless of absence or presence of doxycycline (1µg/ml). In contrast, the inducible melanogenic GLV-1h462 infected groups showed a significantly higher oncolysis rate 48, 72 and 96 hpvi in A549 cells and 72 and 96 hpvi in PANC-1 cells in the absence of dox (***p <0.001; **p <0.01, *p <0.5).
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Figure 3: Oncolytic activity of inducible melanogenic rVACV strains. Photography A) and quantitative evaluation B) of plaque sizes in A594 cancer cells infected with melanogenic rVACV incubated in presence (+) or absence (-) of 1 µg/ml dox. Significantly larger mean maximum plaque diameter of non-induced not melanin producing GLV-1h462 infected A549 cancer cells compared to induced melanogenic GLV-1h462 infected A549 cells or GLV-1h460 infected A549 cancer cells (p < 0.001; n ≥ 50). MTT-Assay of A549 C) and PANC-1 D) cancer cells infected (MOI 0.01 or MOI 0.1) with the constitutively melanin synthesizing GLV-1h460, the non-melanogenic GLV-1h68 and GLV-1h312 and the dox inducible melanogenic GLV-1h462. The control rVACV showed no significant difference in oncolysis, regardless of absence or presence of doxycycline (1µg/ml). In contrast, the inducible melanogenic GLV-1h462 infected groups showed a significantly higher oncolysis rate 48, 72 and 96 hpvi in A549 cells and 72 and 96 hpvi in PANC-1 cells in the absence of dox (***p <0.001; **p <0.01, *p <0.5).

Mentions: Constitutive production of melanin resulted in reduced viral replication as well as decelerated tumor regression when compared to non-melanogenic rVACV 7. To analyze whether we can achieve better replication and spreading in the non-induced state, A549 cells were infected with GLV-1h460 and GLV-1h462 in the presence or absence of 1 µg/ml dox (Fig. 3A and B). Plaques of non-induced and hence not melanin synthesizing GLV-1h462 were bigger than plaques of constitutively melanin producing GLV-1h460 or those of dox-induced melanogenic GLV-1h462 (Fig. 3A). Quantitative analysis of the maximum plaque diameters of each group (Fig. 3B) revealed that the maximum plaque diameters of non-induced not melanin producing GLV-1h462 infected A549 cells were significantly (p < 0.001) bigger (maximum diameter: 124 ± 32µm) than those of induced melanin synthesizing GLV-1h462 infected cells (71 ± 31µm) or GLV-1h460 infected A549 in the absence (67 ± 24µm) or presence (62 ± 22µm) of dox. No significant plaque-diameter difference between melanin producing plaques were observed. Similar results were observed in PANC-1 tumor cells (data not shown). In addition to the enhanced spreading capacity a significantly better oncolytic potential in A549 (Fig. 3C) at 48, 72 and 96 hpvi and in PANC-1 (Fig. 3D) cells at 72 and 96 hpvi of not induced and not melanin-producing GLV-1h462 was demonstrated. Furthermore, a higher replication rate (Supplementary Fig. S1) was observed.


Doxycycline Inducible Melanogenic Vaccinia Virus as Theranostic Anti-Cancer Agent.

Kirscher L, Deán-Ben XL, Scadeng M, Zaremba A, Zhang Q, Kober C, Fehm TF, Razansky D, Ntziachristos V, Stritzker J, Szalay AA - Theranostics (2015)

Oncolytic activity of inducible melanogenic rVACV strains. Photography A) and quantitative evaluation B) of plaque sizes in A594 cancer cells infected with melanogenic rVACV incubated in presence (+) or absence (-) of 1 µg/ml dox. Significantly larger mean maximum plaque diameter of non-induced not melanin producing GLV-1h462 infected A549 cancer cells compared to induced melanogenic GLV-1h462 infected A549 cells or GLV-1h460 infected A549 cancer cells (p < 0.001; n ≥ 50). MTT-Assay of A549 C) and PANC-1 D) cancer cells infected (MOI 0.01 or MOI 0.1) with the constitutively melanin synthesizing GLV-1h460, the non-melanogenic GLV-1h68 and GLV-1h312 and the dox inducible melanogenic GLV-1h462. The control rVACV showed no significant difference in oncolysis, regardless of absence or presence of doxycycline (1µg/ml). In contrast, the inducible melanogenic GLV-1h462 infected groups showed a significantly higher oncolysis rate 48, 72 and 96 hpvi in A549 cells and 72 and 96 hpvi in PANC-1 cells in the absence of dox (***p <0.001; **p <0.01, *p <0.5).
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Related In: Results  -  Collection

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Figure 3: Oncolytic activity of inducible melanogenic rVACV strains. Photography A) and quantitative evaluation B) of plaque sizes in A594 cancer cells infected with melanogenic rVACV incubated in presence (+) or absence (-) of 1 µg/ml dox. Significantly larger mean maximum plaque diameter of non-induced not melanin producing GLV-1h462 infected A549 cancer cells compared to induced melanogenic GLV-1h462 infected A549 cells or GLV-1h460 infected A549 cancer cells (p < 0.001; n ≥ 50). MTT-Assay of A549 C) and PANC-1 D) cancer cells infected (MOI 0.01 or MOI 0.1) with the constitutively melanin synthesizing GLV-1h460, the non-melanogenic GLV-1h68 and GLV-1h312 and the dox inducible melanogenic GLV-1h462. The control rVACV showed no significant difference in oncolysis, regardless of absence or presence of doxycycline (1µg/ml). In contrast, the inducible melanogenic GLV-1h462 infected groups showed a significantly higher oncolysis rate 48, 72 and 96 hpvi in A549 cells and 72 and 96 hpvi in PANC-1 cells in the absence of dox (***p <0.001; **p <0.01, *p <0.5).
Mentions: Constitutive production of melanin resulted in reduced viral replication as well as decelerated tumor regression when compared to non-melanogenic rVACV 7. To analyze whether we can achieve better replication and spreading in the non-induced state, A549 cells were infected with GLV-1h460 and GLV-1h462 in the presence or absence of 1 µg/ml dox (Fig. 3A and B). Plaques of non-induced and hence not melanin synthesizing GLV-1h462 were bigger than plaques of constitutively melanin producing GLV-1h460 or those of dox-induced melanogenic GLV-1h462 (Fig. 3A). Quantitative analysis of the maximum plaque diameters of each group (Fig. 3B) revealed that the maximum plaque diameters of non-induced not melanin producing GLV-1h462 infected A549 cells were significantly (p < 0.001) bigger (maximum diameter: 124 ± 32µm) than those of induced melanin synthesizing GLV-1h462 infected cells (71 ± 31µm) or GLV-1h460 infected A549 in the absence (67 ± 24µm) or presence (62 ± 22µm) of dox. No significant plaque-diameter difference between melanin producing plaques were observed. Similar results were observed in PANC-1 tumor cells (data not shown). In addition to the enhanced spreading capacity a significantly better oncolytic potential in A549 (Fig. 3C) at 48, 72 and 96 hpvi and in PANC-1 (Fig. 3D) cells at 72 and 96 hpvi of not induced and not melanin-producing GLV-1h462 was demonstrated. Furthermore, a higher replication rate (Supplementary Fig. S1) was observed.

Bottom Line: In this study melanin production was detected after exogenous addition of doxycycline in two different tumor xenograft mouse models.Furthermore, it was confirmed that this novel vaccinia virus strain still facilitated signal enhancement as detected by MRI and optoacoustic tomography.At the same time we demonstrated an enhanced oncolytic potential compared to the constitutively melanin synthesizing rVACV system.

View Article: PubMed Central - PubMed

Affiliation: 1. University of Würzburg, Department of Biochemistry, Am Hubland, 97074 Würzburg, Germany.

ABSTRACT
We reported earlier the diagnostic potential of a melanogenic vaccinia virus based system in magnetic resonance (MRI) and optoacoustic deep tissue imaging (MSOT). Since melanin overproduction lead to attenuated virus replication, we constructed a novel recombinant vaccinia virus strain (rVACV), GLV-1h462, which expressed the key enzyme of melanogenesis (tyrosinase) under the control of an inducible promoter-system. In this study melanin production was detected after exogenous addition of doxycycline in two different tumor xenograft mouse models. Furthermore, it was confirmed that this novel vaccinia virus strain still facilitated signal enhancement as detected by MRI and optoacoustic tomography. At the same time we demonstrated an enhanced oncolytic potential compared to the constitutively melanin synthesizing rVACV system.

No MeSH data available.


Related in: MedlinePlus