1. Volume 17, Issue 2, Pages 278-284 (February 2009)
Antiangiogenic Gene Therapy With Soluble VEGFR-1, -2, and -3 Reduces the Growth of Solid Human Ovarian Carcinoma in Mice 
Hanna Sallinen, Maarit Anttila, Johanna Narvainen, Jonna Koponen, Kirsi Hamalainen, Ivana Kholova, Tommi Heikura, Pyry Toivanen, Veli-Matti Kosma, Seppo Heinonen, Kari Alitalo, Seppo Yla-Herttuala 
Molecular Therapy 
Volume 17, Issue 2, Pages (February 2009)
DOI: /mt
Copyright © 2009 The American Society of Gene Therapy Terms and Conditions

2. Figure 1
Protocol of the study and expressions of soluble VEGFRs. (a) Outline of the study. Tumors developed within 3 weeks after the inoculation of the tumor cells. The presence of all tumors was verified by MRI before starting gene therapy. Tumors were observed weekly until the death of the mice. GT, gene therapy; MRI, magnetic resonance imaging. (b) Expression levels of sVEGFR-1, sVEGFR-2, and sVEGFR-3 after adenoviral transduction in SKOV-3m cells were similar as measured by western blotting. (c) sVEGFR-1, sVEGFR-2, and sVEGFR-3 levels in plasma as determined by enzyme-linked immunosorbent assays in combination group VI (sVEGFR-1, sVEGFR-2, and sVEGFR-3). The transgene expression profiles were identical in the other groups. sVEGFR-1, sVEGFR-2, and sVEGFR-3 were not detected in AdLacZ control mice at any time point. Error bars = SEM. (d) Reverse transcription (RT)-PCR was used to confirm the expression of sVEGFR-1, -2, and -3 in mouse liver samples. Lanes 1a–d: sVEGFR-1 RT-PCR. 1a: liver sample 6 days after AdsFlt-1 gene transfer, 1b: liver sample 6 days after AdLacZ gene transfer, 1c: positive control, and 1d: no RT-control. Lanes 2a–d: sVEGFR-3 RT-PCR. 2a: Liver sample 6 days after AdsFlt-4 gene transfer, 2b: liver sample 6 days after AdLacZ gene transfer, 2c: positive control and 2d: no RT-control. Lanes 3a–d: sVEGFR-2 RT-PCR. 3a: liver sample 6 days after AdsKDR gene transfer, 3b: liver sample 6 days after AdLacZ gene transfer, 3c: positive control, and 3d: no RT-control.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2009 The American Society of Gene Therapy Terms and Conditions

3. Figure 2
MRI measurements of tumor growth. (a) Measured by MRI, the mean tumor volumes (mm3) were significantly smaller in group VI (sVEGFR-1, -2, and -3) versus controls 2 weeks after the gene therapy (III MRI) *P < (b) MRI pictures of the development of ovarian tumors in group VI compared to group I (AdLacZ). At the time of the first and the second MRI, there was no difference in tumor volumes, but in the third MRI tumors were significantly smaller in group VI. Tumors are marked with arrows. (c) MRI pictures of the cured mouse in group V (sVEGFR-1 and sVEGFR-3). An intraperitoneal ovarian tumor (arrow) was visible in MRI 18 days after the tumor cell injection. 28 days after the gene therapy, the tumor was shrunken (arrow) and 56 days after the gene therapy the tumor was not visible in MRI. (d) MRI pictures of the mouse in group V (sVEGFR-1 and sVEGFR-3) which had dormancy in tumor growth and a notably prolonged survival. Tumors are marked with arrows. MRI, magnetic resonance imaging.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2009 The American Society of Gene Therapy Terms and Conditions

4. Figure 3
Tumor and ascites measurements. (a) At the end of the follow-up, the weights of the tumors were significantly smaller in both combination groups and in the mice treated with sVEGFR-3. (b) sVEGFR-2-treated mice did not form any ascites in the peritoneal cavity. (c) Microvessel density (MVD: microvessels/mm2) was significantly reduced in mice treated with sVEGFR-2 and with the combination of sVEGFR-1, -2, and -3. (d) combination gene therapy with sVEGFR-1, -2, and -3 significantly reduced the total area of tumors covered by microvessels (TVA: tumor vascular area). *P < 0.05; **P < 0.01; ***P < versus LacZ, Error bars = SEM.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2009 The American Society of Gene Therapy Terms and Conditions

5. Figure 4
Histology of intraperitoneal ovarian tumors. (a) Hematoxylin– eosin staining of serous adenocarcinoma in group I (AdLacZ, left panel). Focal necrosis (arrowhead) and connective tissue (arrow) were present in the tumor tissue in group VI (sVEGFR-1, -2, and -3, right panel). (b) more proliferating tumor cells were seen in control group I (left panel) than in group VI (right panel), 70–80% and 5–40%, respectively. Ki-67 staining. (c) CD-34 positive microvessels in tumor tissue. In group I (left panel), total vascular area was higher than in group VI (right panel). (d) LYVE-1 positive lymphatic vessels were seen in the periphery of tumors in group I (left panel) contrary to the treatment group VI (right panel). Magnification, ×200 (a, b, and d); ×100 (c). Bar = 100 µm.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2009 The American Society of Gene Therapy Terms and Conditions