ABSTRACT
Induced pluripotent stem cell (iPSC) technology offers the promise of immune-matched cell therapies for a wide range of diseases and injuries. It is generally assumed that cells derived from autologous iPSCs will be immune-privileged. However, there are reasons to question this assumption, including recent studies that have tested iPSC immunogenicity in various ways with conflicting results. Understanding the risk of an immune response and developing strategies to minimize it will be important steps before clinical testing. Here, we review the evidence for autologous iPSC immunogenicity, its potential causes, and approaches for assessment and mitigation.
Subject(s)
Induced Pluripotent Stem Cells/immunology , Induced Pluripotent Stem Cells/transplantation , Stem Cell Transplantation , Animals , Clinical Trials as Topic , Humans , Transplantation, AutologousABSTRACT
Cocaine sensitization is produced by repeated exposure to the drug and is thought to reflect neuroadaptations that contribute to addiction. Here, we identify the Ca(2+)/calmodulin-stimulated adenylyl cyclases, type 1 (AC1) and type 8 (AC8), as novel regulators of this behavioral plasticity. We show that, whereas AC1 and AC8 single knock-out mice (AC1(-/-) and AC8(-/-)) exhibit Ca(2+)-stimulated adenylyl cyclase activity in striatal membrane fractions, AC1/8 double-knock-out (DKO) mice do not. Furthermore, DKO mice are acutely supersensitive to low doses of cocaine and fail to display locomotor sensitization after chronic cocaine treatment. Because of the known role for the extracellular signal-regulated kinase (ERK)/mitogen-activated protein kinase signaling pathway in cocaine-induced behavioral plasticity and its coupling to calcium-stimulated cAMP signaling in the hippocampus, we measured phosphorylated ERK (pERK) levels in the striatum. Under basal conditions, pERK is upregulated in choline acetyltransferase-positive interneurons in DKO mice relative to wild-type (WT) controls. After acute cocaine treatment, pERK signaling is significantly suppressed in medium spiny neurons (MSNs) of DKO mice relative to WT mice. In addition to the lack of striatal ERK activation by acute cocaine, signaling machinery downstream of ERK is uncoupled in DKO mice. We demonstrate that AC1 and AC8 are necessary for the phosphorylation of mitogen and stress-activated kinase-1 (pMSK1) at Ser376 and Thr581 and cAMP response element-binding protein (pCREB) at Ser133 after acute cocaine treatment. Our results demonstrate that the Ca(2+)-stimulated adenylyl cyclases regulate long-lasting cocaine-induced behavioral plasticity via activation of the ERK/MSK1/CREB signaling pathway in striatonigral MSNs.
Subject(s)
Adenylyl Cyclases/metabolism , Cocaine/administration & dosage , Dopamine Uptake Inhibitors/administration & dosage , Motor Activity/drug effects , Adenylyl Cyclases/deficiency , Analysis of Variance , Animals , Behavior, Animal/drug effects , CREB-Binding Protein/metabolism , Calcium/pharmacology , Calmodulin/metabolism , Corpus Striatum/drug effects , Corpus Striatum/metabolism , Cyclic AMP-Dependent Protein Kinases/metabolism , Dose-Response Relationship, Drug , Drug Administration Schedule , Extracellular Signal-Regulated MAP Kinases/metabolism , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Motor Activity/physiology , Ribosomal Protein S6 Kinases, 90-kDa/metabolism , Serine/metabolism , Signal Transduction/drug effects , Threonine/metabolism , Up-Regulation/drug effects , Up-Regulation/geneticsABSTRACT
The mitogen-activated protein kinase (MAPK) and cyclic adenosine monophosphate (cAMP) signal transduction pathways have critical roles in the consolidation of hippocampus-dependent memory. We found that extracellular regulated kinase 1/2 MAPK phosphorylation and cAMP underwent a circadian oscillation in the hippocampus that was paralleled by changes in Ras activity and the phosphorylation of MAPK kinase and cAMP response element-binding protein (CREB). The nadir of this activation cycle corresponded with severe deficits in hippocampus-dependent fear conditioning under both light-dark and free-running conditions. Circadian oscillations in cAMP and MAPK activity were absent in memory-deficient transgenic mice that lacked Ca2+ -stimulated adenylyl cyclases. Furthermore, physiological and pharmacological interference with oscillations in MAPK phosphorylation after the cellular memory consolidation period impaired the persistence of hippocampus-dependent memory. These data suggest that the persistence of long-term memories may depend on reactivation of the cAMP/MAPK/CREB transcriptional pathway in the hippocampus during the circadian cycle.
Subject(s)
Circadian Rhythm/physiology , Cyclic AMP/metabolism , Gene Expression Regulation/physiology , Hippocampus/metabolism , Memory/physiology , Mitogen-Activated Protein Kinase Kinases/metabolism , Adenylyl Cyclases/metabolism , Analysis of Variance , Animals , Association Learning/drug effects , Association Learning/physiology , Butadienes/pharmacology , CREB-Binding Protein/metabolism , Calcium/administration & dosage , Conditioning, Classical/drug effects , Conditioning, Classical/physiology , Cyclic AMP/deficiency , Dose-Response Relationship, Drug , Enzyme Inhibitors/pharmacology , Fear , GTP-Binding Proteins/metabolism , Gene Expression Regulation/drug effects , Hippocampus/anatomy & histology , Hippocampus/drug effects , Mice , Mice, Inbred C57BL , Mice, Knockout , Mitogen-Activated Protein Kinase Kinases/deficiency , Motor Activity/physiology , Nitriles/pharmacology , Signal Transduction/physiology , Statistics, Nonparametric , Time FactorsABSTRACT
The cAMP and ERK/MAP kinase (MAPK) signal transduction pathways are critical for hippocampus-dependent memory, a process that depends on CREB-mediated transcription. However, the extent of crosstalk between these pathways and the downstream CREB kinase activated during memory formation has not been elucidated. Here we report that PKA, MAPK, and MSK1, a CREB kinase, are coactivated in a subset of hippocampal CA1 pyramidal neurons following contextual fear conditioning. Activation of PKA, MAPK, MSK1, and CREB is absolutely dependent on Ca(2+)-stimulated adenylyl cyclase activity. We conclude that adenylyl cyclase activity supports the activation of MAPK, and that MSK1 is the major CREB kinase activated during training for contextual memory.
