June 2013
Volume 54, Issue 15
ARVO Annual Meeting Abstract  |   June 2013
SRPK1 Inhibition, As A Way Of Targeting Pro-Angiogenic VEGF-A Production In Ocular Tumours
Author Affiliations & Notes
  • Melissa Gammons
    Department of Physiology and Pharmacology, University of Bristol, Bristol, United Kingdom
  • Sarah Coupland
    Department of Molecular and Clinical Cancer Medicine, University of Liverpool, Liverpool, United Kingdom
  • Andrew Dick
    School of Clinical Sciences and School of Cellular and Molecular medicine, University of Bristol, Bristol, United Kingdom
  • David Bates
    Department of Physiology and Pharmacology, University of Bristol, Bristol, United Kingdom
  • Footnotes
    Commercial Relationships Melissa Gammons, None; Sarah Coupland, None; Andrew Dick, Novartis (C), Novartis (F), GSK (F), Abbott (F); David Bates, University of Bristol (P)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 4526. doi:
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      Melissa Gammons, Sarah Coupland, Andrew Dick, David Bates; SRPK1 Inhibition, As A Way Of Targeting Pro-Angiogenic VEGF-A Production In Ocular Tumours. Invest. Ophthalmol. Vis. Sci. 2013;54(15):4526.

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      © ARVO (1962-2015); The Authors (2016-present)

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Purpose: Uveal melanoma (UM), although rare, is the most common type of intraocular tumour. Approximately 50% of UM are fatal due to their tendency to metastasise to the liver. Angiogenesis is regulated by vascular endothelial growth factor-A (VEGF-A), a growth factor that is expressed by UM. VEGF-A is alternatively spliced to form two families of isoforms: those that are pro-angiogenic and those that are anti-angiogenic. Splicing of the pro-angiogenic isoforms is mediated by serine-rich protein kinase-1 (SRPK1), a protein kinase that results in the phosphorylation and nuclear localization of serine-rich splicing factor 1 (SRSF1). SRSF1 is a promoter of proximal splice site selection, thus selectively upregulating pro-angiogenic VEGF-A production. Activation of this pathway can be achieved by administration of insulin-like growth factor-1 (IGF-1), a promoter of VEGF mediated tumour growth. We investigated the effect of SRPK1 inhibition on VEGF-A splicing in UM cell lines.

Methods: UM cell lines Mel270, Omm2.5 and 92.1 were grown in culture medium. Both before and after 24hours exposure to 1, 5, 10 or 20μM SRPIN340 or combined with 100nM IGF-1 mRNA was extracted, cDNA was synthesised and used in q-PCR and RT-PCR for SRPK1, VEGF165 and VEGF165b expression. Immunofluorescence determined SRSF1 and SRPK1 localisation within the cells, and the protein assessed for VEGF-A expression by Western blot.

Results: SRPK1 was expressed in the UM cytoplasm and translocated to the nucleus during IGF-1 treatment. IGF-1 dose dependently increased pro-angiogenic VEGF-A mRNA expression whereas inhibition of SRPK1 with SRPIN340 reduced pro-angiogenic VEGF-A expression, maintaining the production of anti-angiogenic VEGF-A isoforms.

Conclusions: Targeting SRPK1 reduces the expression of pro-angiogenic VEGF-A in UM cell lines. Selective blocking of pro-angiogenic isoforms may reduce UM metastasis rates whilst maintaining the production of cytoprotective anti-angiogenic isoforms. We wish to validate these results in fresh UM specimens where clinical outcome and genetic status is known.

Keywords: 589 melanoma • 744 tumors • 748 vascular endothelial growth factor  

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