title: Regulation of alternative VEGF-A mRNA splicing is a therapeutic target for analgesia.
abstract: Vascular endothelial growth factor-A (VEGF-A) is best known as a key regulator of the formation of new blood vessels. Neutralization of VEGF-A with anti-VEGF therapy e.g. bevacizumab, can be painful, and this is hypothesized to result from a loss of VEGF-A-mediated neuroprotection. The multiple vegf-a gene products consist of two alternatively spliced families, typified by VEGF-A165a and VEGF-A165b (both contain 165 amino acids), both of which are neuroprotective. Under pathological conditions, such as in inflammation and cancer, the pro-angiogenic VEGF-A165a is upregulated and predominates over the VEGF-A165b isoform. We show here that in rats and mice VEGF-A165a and VEGF-A165b have opposing effects on pain, and that blocking the proximal splicing event - leading to the preferential expression of VEGF-A165b over VEGF165a - prevents pain in vivo. VEGF-A165a sensitizes peripheral nociceptive neurons through actions on VEGFR2 and a TRPV1-dependent mechanism, thus enhancing nociceptive signaling. VEGF-A165b blocks the effect of VEGF-A165a. After nerve injury, the endogenous balance of VEGF-A isoforms switches to greater expression of VEGF-Axxxa compared to VEGF-Axxxb, through an SRPK1-dependent pre-mRNA splicing mechanism. Pharmacological inhibition of SRPK1 after traumatic nerve injury selectively reduced VEGF-Axxxa expression and reversed associated neuropathic pain. Exogenous VEGF-A165b also ameliorated neuropathic pain. We conclude that the relative levels of alternatively spliced VEGF-A isoforms are critical for pain modulation under both normal conditions and in sensory neuropathy. Altering VEGF-Axxxa/VEGF-Axxxb balance by targeting alternative RNA splicing may be a new analgesic strategy.
authors.affiliation: Physiology and Pharmacology, University of Bristol, Bristol BS8 1TD, UK; Cancer Biology, Division of Cancer and Stem Cells, School of Medicine, University of Nottingham, Queen's Medical Centre, Nottingham NG2 7UH, UK.
authors.forename: R P
authors.initials: RP
authors.lastname: Hulse
authors.affiliation: Physiology and Pharmacology, University of Bristol, Bristol BS8 1TD, UK; School of Life Sciences, The Medical School, University of Nottingham, Queen's Medical Centre, Nottingham NG2 7UH, UK.
authors.forename: N
authors.initials: N
authors.lastname: Beazley-Long
authors.affiliation: Physiology and Pharmacology, University of Bristol, Bristol BS8 1TD, UK.
authors.forename: J
authors.initials: J
authors.lastname: Hua
authors.affiliation: Physiology and Pharmacology, University of Bristol, Bristol BS8 1TD, UK.
authors.forename: H
authors.initials: H
authors.lastname: Kennedy
authors.affiliation: Physiology and Pharmacology, University of Bristol, Bristol BS8 1TD, UK.
authors.forename: J
authors.initials: J
authors.lastname: Prager
authors.affiliation: Physiology and Pharmacology, University of Bristol, Bristol BS8 1TD, UK.
authors.forename: H
authors.initials: H
authors.lastname: Bevan
authors.affiliation: Physiology and Pharmacology, University of Bristol, Bristol BS8 1TD, UK.
authors.forename: Y
authors.initials: Y
authors.lastname: Qiu
authors.affiliation: King's College London, London SE1 9NH, UK.
authors.forename: E S
authors.initials: ES
authors.lastname: Fernandes
authors.affiliation: Physiology and Pharmacology, University of Bristol, Bristol BS8 1TD, UK.
authors.forename: M V
authors.initials: MV
authors.lastname: Gammons
authors.affiliation: Paul Scherrer Institut, 5232 Villigen, Switzerland.
authors.forename: K
authors.initials: K
authors.lastname: Ballmer-Hofer
authors.affiliation: Anatomy and Embryology, Leiden University Medical Centre, 2300 RC Leiden, The Netherlands.
authors.forename: A C
authors.initials: AC
authors.lastname: Gittenberger de Groot
authors.affiliation: Clinical Sciences, University of Bristol, Bristol BS1 2LX, UK.
authors.forename: A J
authors.initials: AJ
authors.lastname: Churchill
authors.affiliation: Physiology and Pharmacology, University of Bristol, Bristol BS8 1TD, UK.
authors.forename: S J
authors.initials: SJ
authors.lastname: Harper
authors.affiliation: King's College London, London SE1 9NH, UK.
authors.forename: S D
authors.initials: SD
authors.lastname: Brain
authors.affiliation: Cancer Biology, Division of Cancer and Stem Cells, School of Medicine, University of Nottingham, Queen's Medical Centre, Nottingham NG2 7UH, UK. Electronic address: David.Bates@nottingham.ac.uk.
authors.forename: D O
authors.initials: DO
authors.lastname: Bates
authors.affiliation: Physiology and Pharmacology, University of Bristol, Bristol BS8 1TD, UK; School of Life Sciences, The Medical School, University of Nottingham, Queen's Medical Centre, Nottingham NG2 7UH, UK. Electronic address: Lucy.Donaldson@nottingham.ac.uk.
authors.forename: L F
authors.initials: LF
authors.lastname: Donaldson
mesh_terms: D000818:Animals; D000906:Antibodies / Q000494:pharmacology / Q000627:therapeutic use*; D001572:Benzofurans; D004274:DNA, Recombinant / Q000235:genetics*; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000494:pharmacology / Q000627:therapeutic use; D005727:Ganglia, Spinal / Q000166:cytology; D006930:Hyperalgesia / Q000378:metabolism; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009431:Neural Conduction / Q000235:genetics; D009437:Neuralgia / Q000378:metabolism* / Q000628:therapy*; D010147:Pain Measurement; D017288:Pain Threshold / Q000502:physiology; D011804:Quinolines; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D011984:Sensory Receptor Cells / Q000187:drug effects / Q000378:metabolism; D050916:TRPV Cation Channels / Q000172:deficiency / Q000235:genetics; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000276:immunology / Q000378:metabolism
chemical_list: C576715:(3aS,4S,9bS)-N-(2-(8-cyano-1-formyl-2,3,3a,4,5,9b-hexahydro-1H-pyrrolo(3,2-c)quinolin-4-yl)-2-methylpropyl)-4,6-difluorobenzofuran-2-carboxyamide; D000906:Antibodies; D001572:Benzofurans; D004274:DNA, Recombinant; D004791:Enzyme Inhibitors; D011804:Quinolines; D012333:RNA, Messenger; D050916:TRPV Cation Channels; C493921:TRPV1 protein, mouse; D042461:Vascular Endothelial Growth Factor A
keywords: Alternative mRNA splicing; Neuropathy; Nociceptors; Vascular endothelial growth factor A