Identification of compounds that potentiate CREB signaling as possible enhancers of long-term memory

Many studies have implicated the cAMP Response Element Binding (CREB) protein signaling pathway in long-term memory. To identify small molecule enhancers of CREB activation of gene expression, we screened ≈73,000 compounds, each at 7–15 concentrations in a quantitative high-throughput screening (qHTS) format, for activity in cells by assaying CREB mediated β-lactamase reporter gene expression. We identified 1,800 compounds that potentiated CREB mediated gene expression, with potencies as low as 16 nM, comprising 96 structural series. Mechanisms of action were systematically determined, and compounds that affect phosphodiesterase 4, protein kinase A, and cAMP production were identified, as well as compounds that affect CREB signaling via apparently unidentified mechanisms. qHTS folowed by interrogation of pathway targets is an efficient paradigm for lead generation for chemical genomics and drug development.

[1]  Raoul C. M. Hennekam,et al.  Rubinstein-Taybi syndrome caused by mutations in the transcriptional co-activator CBP , 1995, Nature.

[2]  J. Casida,et al.  Protein phosphatase 2A and its [3H]cantharidin/[3H]endothall thioanhydride binding site. Inhibitor specificity of cantharidin and ATP analogues. , 1993, Biochemical pharmacology.

[3]  T. Kitamoto,et al.  A clock gene, period, plays a key role in long-term memory formation in Drosophila. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[4]  S. Hess,et al.  1-Ethyl-4-(isopropylidenehydrazino)-1H-pyrazolo-(3,4-b)-pyridine-5-carboxylic acid, ethyl ester, hydrochloride (SQ 20009)--a potent new inhibitor of cyclic 3',5'-nucleotide phosphodiesterases. , 1972, Biochemical pharmacology.

[5]  M. Montminy,et al.  Phosphorylation-induced binding and transcriptional efficacy of nuclear factor CREB , 1988, Nature.

[6]  W. Quinn,et al.  Induction of a dominant negative CREB transgene specifically blocks long-term memory in Drosophila , 1994, Cell.

[7]  H. Inoue,et al.  Acute and chronic effects of T-1032, a novel selective phosphodiesterase type 5 inhibitor, on monocrotaline-induced pulmonary hypertension in rats. , 2002, Biological & pharmaceutical bulletin.

[8]  E. Kandel,et al.  Activation of cAMP-Responsive genes by stimuli that produce long-term facilitation in aplysia sensory neurons , 1993, Neuron.

[9]  Alcino J. Silva,et al.  Blockade of cyclic AMP‐responsive element DNA binding in the brain of CREBΔ/α mutant mice , 2000, Neuroreport.

[10]  Richard Morphy,et al.  Fragments, network biology and designing multiple ligands. , 2007, Drug discovery today.

[11]  Adam Yasgar,et al.  Quantitative high-throughput screening: a titration-based approach that efficiently identifies biological activities in large chemical libraries. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[12]  A. Means,et al.  Calcium/calmodulin-dependent protein kinase types II and IV differentially regulate CREB-dependent gene expression , 1994, Molecular and cellular biology.

[13]  E. Kandel,et al.  Chromatin Acetylation, Memory, and LTP Are Impaired in CBP+/− Mice A Model for the Cognitive Deficit in Rubinstein-Taybi Syndrome and Its Amelioration , 2004, Neuron.

[14]  G. Livera,et al.  Cyclic AMP-specific PDE4 Phosphodiesterases as Critical Components of Cyclic AMP Signaling* , 2003, The Journal of Biological Chemistry.

[15]  A. Shaywitz,et al.  CREB: a stimulus-induced transcription factor activated by a diverse array of extracellular signals. , 1999, Annual review of biochemistry.

[16]  Alcino J. Silva,et al.  Deficient long-term memory in mice with a targeted mutation of the cAMP-responsive element-binding protein , 1994, Cell.

[17]  H Terada,et al.  A High-Efficiency Protein Transduction System Demonstrating the Role of PKA in Long-Lasting Long-Term Potentiation , 2001, The Journal of Neuroscience.

[18]  J. Beavo,et al.  Cloning and characterization of a cAMP-specific cyclic nucleotide phosphodiesterase. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[19]  M. Montminy,et al.  Cyclic AMP stimulates somatostatin gene transcription by phosphorylation of CREB at serine 133 , 1989, Cell.

[20]  C. Lugnier.  Cyclic nucleotide phosphodiesterase (PDE) superfamily: a new target for the development of specific therapeutic agents. , 2006, Pharmacology & therapeutics.

[21]  Christina R. Maxwell,et al.  Phosphodiesterase inhibitors: A novel mechanism for receptor-independent antipsychotic medications , 2004, Neuroscience.

[22]  E. Nestler,et al.  The many faces of CREB , 2005, Trends in Neurosciences.

[23]  Christopher P Austin,et al.  High-throughput screening assays for the identification of chemical probes. , 2007, Nature chemical biology.

[24]  D. Pereira,et al.  Origin and evolution of high throughput screening , 2007, British journal of pharmacology.

[25]  G L Johnson,et al.  Nuclear protein phosphatase 2A dephosphorylates protein kinase A-phosphorylated CREB and regulates CREB transcriptional stimulation , 1993, Molecular and cellular biology.

[26]  R. Owens,et al.  Development of a recombinant cell-based system for the characterisation of phosphodiesterase 4 isoforms and evaluation of inhibitors. , 1999, Biochemical pharmacology.

[27]  K. Fujishige,et al.  Characterization and effects of methyl-2- (4-aminophenyl)-1, 2-dihydro-1-oxo-7- (2-pyridinylmethoxy)-4-(3,4, 5-trimethoxyphenyl)-3-isoquinoline carboxylate sulfate (T-1032), a novel potent inhibitor of cGMP-binding cGMP-specific phosphodiesterase (PDE5). , 2000, Biochemical pharmacology.

[28]  M. Nirenberg,et al.  Identification of a potent new chemotype for the selective inhibition of PDE4. , 2008, Bioorganic & medicinal chemistry letters.

[29]  K. Jarnagin,et al.  Comparison of recombinant human PDE4 isoforms: interaction with substrate and inhibitors. , 1998, Cellular signalling.

[30]  O. Vitolo,et al.  Persistent improvement in synaptic and cognitive functions in an Alzheimer mouse model after rolipram treatment. , 2004, The Journal of clinical investigation.

[31]  Masatoshi Hagiwara,et al.  Phosphorylated CREB binds specifically to the nuclear protein CBP , 1993, Nature.

[32]  M. Hagiwara,et al.  Transcriptional attenuation following cAMP induction requires PP-1-mediated dephosphorylation of CREB , 1992, Cell.

[33]  C. McMurray,et al.  Calmodulin Kinase II Attenuation of Gene Transcription by Preventing cAMP Response Element-binding Protein (CREB) Dimerization and Binding of the CREB-binding Protein* , 2001, The Journal of Biological Chemistry.

[34]  T. Tully,et al.  A mouse model of Rubinstein-Taybi syndrome: Defective long-term memory is ameliorated by inhibitors of phosphodiesterase 4 , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[35]  Ronald L. Davis,et al.  The Role of cAMP Response Element-Binding Protein in Drosophila Long-Term Memory , 2004, The Journal of Neuroscience.

[36]  J. Kotera,et al.  Overview of PDEs and Their Regulation , 2007, Circulation research.