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VEGF121 and VEGF165 differentially promote vessel maturation and tumor growth in mice and humans

Abstract

Tumor angiogenesis depends on the vascular endothelial growth factor (VEGF), which exists in multiple splicing isoforms, including the most abundant VEGF165 and VEGF121. We have previously shown that the differential capacity of these two VEGF isoforms to bind Neuropilin-1 accounts for their diverse ability to recruit Nrp1-expressing monocytes (NEMs), resulting in a different arteriogenic potential. Here we measure the expression of VEGF165 and VEGF121 in human cancer and their influence on tumor growth and vascularization. We measured the expression levels of VEGF165 and VEGF121 in human colorectal cancer and found that VEGF121 was more expressed than VEGF165, particularly in patients with extensive lymph node infiltration. Overexpressing either VEGF165 or VEGF121 in a cancer mouse model, we observed that the former decreased, whereas the latter increased tumor growth. In both clinical and experimental tumors, VEGF165 expression resulted in the recruitment of NEMs, paralleled by maturation of the tumor vascular network. Finally, hypoxia induced a shift toward the VEGF165 isoform in the central core of human cancers, as well as in various types of cultured cells. These results demonstrate that the two VEGF splicing isoforms are differentially expressed in colorectal cancers, exerting opposite effects on tumor growth and vessel maturation.

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References

  1. Giacca M, Zacchigna S . VEGF gene therapy: therapeutic angiogenesis in the clinic and beyond. Gene Ther 2012; 19: 622–629.

    Article  CAS  Google Scholar 

  2. Ferrara N, Gerber HP, LeCouter J . The biology of VEGF and its receptors. Nat Med 2003; 9: 669–676.

    Article  CAS  Google Scholar 

  3. Zacchigna S, Pattarini L, Zentilin L, Moimas S, Carrer A, Sinigaglia M et al. Bone marrow cells recruited through the Neuropilin-1 receptor promote arterial formation at the sites of adult neoangiogenesis. J Clin Invest 2008; 118: 2062–2075.

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Carrer A, Moimas S, Zacchigna S, Pattarini L, Zentilin L, Ruozi G et al. Neuropilin-1 identifies a subset of bone marrow Gr1- monocytes that can induce tumor vessel normalization and inhibit tumor growth. Cancer Res 2012; 72: 6371–6381.

    Article  CAS  Google Scholar 

  5. Misquitta-Ali CM, Cheng E, O'Hanlon D, Liu N, McGlade CJ, Tsao MS et al. Global profiling and molecular characterization of alternative splicing events misregulated in lung cancer. Mol Cell Biol 2011; 31: 138–150.

    Article  CAS  Google Scholar 

  6. Zhao YJ, Han HZ, Liang Y, Shi CZ, Zhu QC, Yang J . Alternative splicing of VEGFA, APP and NUMB genes in colorectal cancer. World J Gastroenterol 2015; 21: 6550–6560.

    Article  CAS  Google Scholar 

  7. Tomayko MM, Reynolds CP . Determination of subcutaneous tumor size in athymic (nude) mice. Cancer Chemother Pharmacol 1989; 24: 148–154.

    Article  CAS  Google Scholar 

  8. Arsic N, Zentilin L, Zacchigna S, Santoro D, Stanta G, Salvi S et al. Induction of functional neovascularization by combined VEGF and angiopoietin-1 gene transfer using AAV vectors. Mol Ther 2003; 7: 450–459.

    Article  CAS  Google Scholar 

  9. Miles AA, Miles EM . Vascular reaction to histamine, histamine-liberator, and leukotaxine in the skin of guinea pigs. J Physiol 1952; 118: 228–257.

    Article  CAS  Google Scholar 

  10. Zentilin L, Giacca M . Competitive PCR for precise nucleic acid quantification. Nat Protoc 2007; 2: 2092–2104.

    Article  CAS  Google Scholar 

  11. Yoon C, Kim D, Kim S, Park GB, Hur DY, Yang JW et al. MiR-9 regulates the post-transcriptional level of VEGF165a by targeting SRPK-1 in ARPE-19 cells. Graefes Arch Clin Exp Ophthalmol 2014; 252: 1369–1376.

    Article  CAS  Google Scholar 

  12. Ferrara N . Binding to the extracellular matrix and proteolytic processing: two key mechanisms regulating vascular endothelial growth factor action. Mol Biol Cell 2010; 21: 687–690.

    Article  CAS  Google Scholar 

  13. Berkman RA, Merrill MJ, Reinhold WC, Monacci WT, Saxena A, Clark WC et al. Expression of the vascular permeability factor/vascular endothelial growth factor gene in central nervous system neoplasms. J Clin Invest 1993; 91: 153–159.

    Article  CAS  Google Scholar 

  14. Cai C, Bottcher MC, Werner JA, Mandic R . Differential expression of VEGF121, VEGF165 and VEGF189 in angiomas and squamous cell carcinoma cell lines of the head and neck. Anticancer Res 2010; 30: 805–810.

    CAS  PubMed  Google Scholar 

  15. Levy AP, Levy NS, Loscalzo J, Calderone A, Takahashi N, Yeo KT et al. Regulation of vascular endothelial growth factor in cardiac myocytes. Circ Res 1995; 76: 758–766.

    Article  CAS  Google Scholar 

  16. Salton M, Voss TC, Misteli T . Identification by high-throughput imaging of the histone methyltransferase EHMT2 as an epigenetic regulator of VEGFA alternative splicing. Nucleic Acids Res 2014; 42: 13662–13673.

    Article  CAS  Google Scholar 

  17. Elias AP, Dias S . Microenvironment Changes (in pH) Affect VEGF Alternative Splicing. Cancer Microenviron 2008; 1: 131–139.

    Article  Google Scholar 

  18. Carmeliet P, Jain RK . Principles and mechanisms of vessel normalization for cancer and other angiogenic diseases. Nat Rev Drug Discov 2011; 10: 417–427.

    Article  CAS  Google Scholar 

  19. Maione F, Molla F, Meda C, Latini R, Zentilin L, Giacca M et al. Semaphorin 3 A is an endogenous angiogenesis inhibitor that blocks tumor growth and normalizes tumor vasculature in transgenic mouse models. J Clin Invest 2009; 119: 3356–3372.

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Van de Veire S, Stalmans I, Heindryckx F, Oura H, Tijeras-Raballand A, Schmidt T et al. Further pharmacological and genetic evidence for the efficacy of PlGF inhibition in cancer and eye disease. Cell 2010; 141: 178–190.

    Article  CAS  Google Scholar 

  21. Woolard J, Wang WY, Bevan HS, Qiu Y, Morbidelli L, Pritchard-Jones RO et al. VEGF165b, an inhibitory vascular endothelial growth factor splice variant: mechanism of action, in vivo effect on angiogenesis and endogenous protein expression. Cancer Res 2004; 64: 7822–7835.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We are grateful to Stefano Artico and Barbara Boziglav for excellent technical support. This work was supported by Associazione Italiana per la Ricerca sul Cancro (AIRC, MFAG n. 9233) to S. Zacchigna and Italian Ministry of Education, University and Research (MIUR), FIERCE project n. RBAP11Z4Z9 to M. Giacca.

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Correspondence to S Zacchigna.

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Supplementary Information accompanies the paper on Cancer Gene Therapy website

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Kazemi, M., Carrer, A., Moimas, S. et al. VEGF121 and VEGF165 differentially promote vessel maturation and tumor growth in mice and humans. Cancer Gene Ther 23, 125–132 (2016). https://doi.org/10.1038/cgt.2016.12

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