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Pharmacology of a novel TRPM2 antagonist, JNJ‐28583113
Author(s) -
Fourgeaud Lawrence,
Faouzi Malika,
Starkus John,
Wang Qi,
Coates Heather,
He Yingbo,
Lovenberg Timothy,
Penner Reinhold,
Dvorak Curt,
Bhattacharya Anindya
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.1188.1
Subject(s) - trpm2 , microglia , calcium in biology , chemistry , microbiology and biotechnology , neuroinflammation , biology , pharmacology , intracellular , receptor , biochemistry , transient receptor potential channel , immunology , inflammation
TRPM2 is a calcium‐permeable non‐selective cation channel that is activated, in a synergistic manner, by both intracellular adenosine diphosphate ribose (ADPR) and calcium. It is also activated indirectly by reactive oxygen nitrogen species (ROS/NOS) due to increased ADPR production during oxidative/nitrosative stress. TRPM2 is primarily expressed in the immune cells and central nervous system (CNS) where it plays important roles in pathologies involving neuroimmune interactions, particularly in the context of oxidative stress. Within the CNS, TRPM2 is expressed mainly in neurons and microglia. Here, we present the pharmacological properties of JNJ‐28583113, a newly discovered compound representing the most potent TRPM2 antagonist described to date. Our data show that this compound has a potency range of 100–200 nM in blocking H 2 O 2 ‐induced calcium flux in several recombinant systems over‐expressing human, chimpanzee and rat TRPM2. In whole‐cell patch clamp recordings, JNJ‐285831113 blocked intracellular ADPR‐induced human TRPM2 currents with a potency of 13 ± 3 nM. Similar pharmacology was observed for native TRPM2 in a human macrophage cell line (U937) and in a rat pancreatic cell line (INS‐1). In line with TRPM2 microglial expression, JNJ‐28583113 also modulated lipopolysaccharide (LPS)‐induced changes gliotransmitter secretion in mouse microglia in vitro , indicating a possible role of this channel in neuroinflammation. Using multi‐electrode arrays to record hippocampal field potentials in acute mouse brain slices, we observed that JNJ‐285831113 prevented hypoxia‐induced synaptic depression at CA3 to CA1 synapses, suggesting a role for TRPM2 in synaptic changes occurring during oxidative stress. Finally, JNJ‐28583113 inhibited H 2 O 2 ‐induced dephosphorylation of glycogen synthase kinase‐3 (GSK3) alpha and beta in recombinant cells over‐expressing human TRPM2 channels, confirming the importance of TRPM2 in modulating GSK3 signaling. Together our results demonstrate that JNJ‐28583113 is a very potent TRPM2 antagonist that can be used to further elucidate TRPM2 physiological functions as well as to probe the role of TRPM2 in disease biology under conditions of oxidative stress. Support or Funding Information Janssen