AMPA receptor trafficking in the pathophysiology and treatment of mood disorders

AMPA 受体转运在情绪障碍的病理生理学和治疗中的作用

• 批准号: 8158114
• 负责人: MILES A. HERKENHAM
• 金额: $ 39.56万
• 依托单位: NATIONAL INSTITUTE OF MENTAL HEALTH
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8158114
• 关键词: AMPA Receptors; Functional disorder; Mood Disorders; trafficking

Mood disorders are one of the leading causes of disability worldwide, ranking ahead of ischemic heart disease, cerebrovascular disease, cancers, and infectious diseases. Despite the devastating impact that mood disorders have on the lives of millions of individuals worldwide, little is known about their underlying etiology and neurobiology. Currently available treatments are insufficient for many patients suffering from mood disorders, and even for those patients who do respond to available antidepressants or mood stabilizers, there is a significant therapeutic lag before clinical benefits appear. Notably, to date, no drug has been developed specifically to treat bipolar disorder (BD) based on an understanding of the neurobiological basis of the illness or the mechanism of action of existing effective medications. Thus, the major challenge in BD research is to find a common, convergent, functional mechanism associated with BD in order to develop urgently needed and truly novel and effective therapeutics. In this study, we sought to identify this common, convergent system in mood disorders and to develop a new potential drug that mimics the effect of mood stabilizers on GluR1 phosphorylation. TAT-peptides (TAT-p845 and TAT-SRC) were designed and synthesized. TAT peptide (YGRKKRRQRRR) linked to the functional peptide enables the functional peptide to pass through the blood-brain barrier and cell membrane, allowing it to get into cytosol or synapses of the neurons. A previous study has successfully utilized TAT peptide injection into animals to disrupt the interaction of the postsynaptic density protein PSD-95 with NMDA receptors in the brain and to provide a neuroprotective effect on a stroke animal model. TAT-p845 was able to inhibit the phosphorylation of AMPA receptors at the PKA site and down-regulate the surface expression of GluR1 in cultured hippocampal neurons, which is the same effect produced by lithium and valproate. Moreover, this TAT-p845 was able to pass the blood-brain barrier and inhibit the phosphorylation of GluR1 in the hippocampus in vivo, which again demonstrated its ability to induce the same effects as lithium and valproate. In addition, reduction of GluR1 phosphorylation at its PKA site by TAT-p845 was sufficient to attenuate synaptic GluR1/2 in hippocampal neurons in vivo. Intra-hippocampal infusion of AMPA-specific inhibitor GYKI54226, GluR1-specific TAT-p845 peptide, and GluR1-PDZ-specific TAT-TGL peptide were able to attenuate amphetamine-induced hyperactivity and/or amphetamine-induced conditioned-place preference in the mania animal model. These studies provide novel mechanisms for anti-manic effect through attenuation of AMPA receptor activity and suggest avenues for new drug development for mood disorders. TAT-p845, which attenuates AMPA receptor levels at synapses, may offer exciting possibilities as a new class of medicine with the potential for treatment of bipolar disorder. Considerable biochemical evidence suggests that the protein kinase C (PKC) signaling cascade may be an important pathway for the actions of anti-manic agents, and that excessive PKC activation can disrupt prefrontal cortical regulation of thinking and behavior. Currently, however, brain protein targets of PKCs anti-manic effects remain unclear. Based on a previous finding, we want to determine how PKC-mediated regulation of glutamate receptors plays an important role in the pathophysiology and treatment of mania. Here we showed that PKC activity was enhanced in the prefrontal cortex of animals treated with the psychostimulant amphetamine and the antidepressant imipramine. Phosphorylation of myristoylated alanine-rich C kinase substrate (MARCKS), a marker of PKC activity, was increased in the prefrontal cortex of psychostimulant-treated animals, as well as in sleep-deprived animals (another animal model of mania), but decreased in lithium-treated animals. The antidepressant imipramine, which shows promanic property on bipolar patients, also enhanced pMARCKS in prefrontal cortex in vivo. We further explored the functional targets of PKC in mania-associated behaviors. Neurogranin is a brain-specific, postsynaptically located PKC substrate. PKC phosphorylation of neurogranin was robustly increased by pro-manic manipulations and decreased by the anti-manic agent. PKC phosphorylation of the NMDA receptor site NR1S896 and the AMPA receptor site GluR1T840 was also enhanced in the prefrontal cortex of animals treated with antidepressant imipramine, as well as behaviorally sleep-deprived, in striking contrast to the reduced activity seen in lithium-treated animals. These results suggest that PKC may play an important role in regulating NMDA and AMPA receptor functions. The biochemical profile of the PKC pathway thus encompasses both pro- and anti-manic effects on behavior. These results suggest that PKC modulators or their intracellular targets may ultimately represent novel avenues for the development of new therapeutics for mood disorders. To gain mechanistic insight into the synaptic and behavioral changes associated with GluR1 deletion, hippocampal genome-wide expression profiling was conducted using groups of GluR1 knockout (KO) mice and their wild-type littermates. Regulation of 38 genes was found to be altered more than 30% (P < 0.01, n = 8), and seven of these genes were studied with additional quantitative experiments. A large portion of the altered genes encoded molecules involved in calcium signaling, including calcium channel components, calcium-binding proteins and calcium-calmodulin-dependent protein kinase II subunits. At the protein level, we further evaluated some genes in the calcium pathway that were altered in GluR1 KO mice. Protein levels of two key molecules in the calcium pathway - GluR, ionotropic, N-methyl-d-aspartate-1 and calcium/calmodulin-dependent protein kinase II alpha - showed similar changes to those observed in mRNA levels. These findings raise the possibility that calcium signaling and other plasticity molecules may contribute to the hippocampal plasticity and behavioral deficits observed in GluR1 KO mice. Recent genome-wide association studies (GWAS) have made progress in identifying genetic markers that are reproducibly associated with BD. However, these markers explain very little of the inherited risk of BD and the implicated genets bear no obvious pathophysiological relationship to the disease. Here we sought to identify etiologic pathways for BD by examining genes that lie near each of the top 500 SNPs associated with BD in a GWAS meta-analysis that comprised about 12000 cases and controls. We classified the genes into functional groups, based on the published literature, then compared this grouping with that obtained by DAVID, a widely-used data mining tool. As a comparison group, we used the results of a large GWAS meta-analysis of height, a classic polygenic trait. Genes that lay near SNPs associated with BD were overrepresented in the cytoskeleton interaction gene cluster. Most genes in this cluster modulate synaptic strength by regulating vesicle trafficking and structural stability of the synapses. Recent pharmacological studies have shown that synaptic strength may be involved in the pathophysiology and treatment of BD. DAVID analysis also detected a significant over-representation of these within the cytoskeleton interaction and synapses groups. Overall, we found that about 50% of BD associated SNPS lie near genes whose known functions directly regulate synaptic strength. We detected no significant over-representation of these gene groups in the height data. We conclude that the human genetic data from GWAS support the hypothesis from pharmacological research that synaptic strength is involved in the etiology of BD.

项目成果

Genome-wide gene expression profiling in GluR1 knockout mice: key role of the calcium signaling pathway in glutamatergically mediated hippocampal transmission.

GLUR1基因敲除小鼠中全基因组基因表达分析：谷氨酸能介导的海马传播中钙信号通路的关键作用。

• DOI: 10.1111/j.1460-9568.2009.07022.x
• 发表时间: 2009-12
• 期刊: The European journal of neuroscience
• 作者: Zhou R;Holmes A;Du J;Malkesman O;Yuan P;Wang Y;Damschroder-Williams P;Chen G;Guitart X;Manji HK
• 通讯作者: Manji HK