Lysophosphatidic acid-induced platelet shape change revealed through LPA_1–5 receptor-selective probes and albumin

Abstract

Lysophosphatidic acid (LPA), a component of mildly-oxidized LDL and the lipid rich core of atherosclerotic plaques, elicits platelet activation. LPA is the ligand of G protein-coupled receptors (GPCR) of the EDG family (LPA_1–3) and the newly identified LPA_4–7 subcluster. LPA₄, LPA₅ and LPA₇ increase cellular cAMP levels that would induce platelet inhibition rather than activation. In the present study we quantified the mRNA levels of the LPA_1–7 GPCR in human platelets and found a rank order LPA₄ = LPA₅ > LPA₇ > LPA₆ = LPA₂ ≫ LPA₁ > LPA₃. We examined platelet shape change using a panel of LPA receptor subtype-selective agonists and antagonists and compared them with their pharmacological profiles obtained in heterologous LPA_1–5 receptor expression systems. Responses to different natural acyl and alkyl species of LPA, and octyl phosphatidic acid analogs, alpha-substituted phosphonate analogs, N-palmitoyl-tyrosine phosphoric acid, N-palmitoyl-serine phosphoric acid were tested. All of these compounds elicited platelet activation and also inhibited LPA-induced platelet shape change after pre-incubation, suggesting that receptor desensitization is likely responsible for the inhibition of this response. Fatty acid free albumin (10 µM) lacking platelet activity completely inhibited platelet shape change induced by LPA with an IC₅₀ of 1.1 µM but had no effect on the activation of LPA_1,2,3,&5 expressed in endogenously non-LPA-responsive RH7777 cells. However, albumin reduced LPA₄ activation and shifted the dose-response curve to the right. LPA₅ transiently expressed in RH7777 cells showed preference to alkyl-LPA over acyl-LPA that is similar to that in platelets. LPA did not increase cAMP levels in platelets. In conclusion, our results with the pharmacological compounds and albumin demonstrate that LPA does not induce platelet shape change simply through activation of LPA_1–5, and the receptor(s) mediating LPA-induced platelet activation remains elusive.

• Lysophosphatidic acid
• G protein-coupled receptor
• agonist
• antagonist
• albumin