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Expression of EGF receptor and its signals in HB-EGF KO and control mice. Representative images of immunoblots showing EGF receptor and b-actin (a), ErbB4 and b-actin (b), p-EGF receptor and total EGF receptor (c), p-ERK, ERK, and b-actin (d), p-Akt, Akt, and b-actin (e), and p-ErbB4 and ErbB4 (f) in the prefrontal cortex (upper). Quantitative analysis of EGF receptor (a), ErbB4 (b), p-EGF receptor (c), p-ERK (d), p-Akt (e), and p-ErbB4 (f) by densitometric scanning of immunoreactive bands, with normalization to the signals for b-actin (a and b), EGF receptor (c), ERK (d), Akt (e), or ErbB4 (f) (lower). Data for control (n = 5) and KO (n = 5) mice are expressed as a percentage of the control value. Values are means6SEM. * p,0.05 vs. control mice. doi:10.1371/journal.pone.0007461.g006
Source publication
Recently, neurotrophic factors and cytokines have been shown to be associated in psychiatric disorders, such as schizophrenia, bipolar disorder, and depression. Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a member of the EGF family, serves as a neurotrophic molecular and plays a significant role in the brain. We generated...
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... that might lead to the observed behavioral abnormalities and altered monoamine levels, we compared the expression of the EGF receptor and its signaling in HB-EGF KO and control mice. No significant differences in the expression of total EGF receptor or ErbB4 proteins were observed between controls and HB-EGF KO mice (P.0.05 vs. control mice) ( Fig. 6a and b). Despite these equivalent levels of EGF receptor expression, p-EGF receptor protein was significantly decreased in the prefrontal cortex of HB-EGF KO mice (P,0.05 vs. control mice) (Fig. 6c). Moreover, the expression of phosphorylated extracellular signal-regulated kinase (p-ERK) protein was significantly decreased in the prefrontal ...
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... differences in the expression of total EGF receptor or ErbB4 proteins were observed between controls and HB-EGF KO mice (P.0.05 vs. control mice) ( Fig. 6a and b). Despite these equivalent levels of EGF receptor expression, p-EGF receptor protein was significantly decreased in the prefrontal cortex of HB-EGF KO mice (P,0.05 vs. control mice) (Fig. 6c). Moreover, the expression of phosphorylated extracellular signal-regulated kinase (p-ERK) protein was significantly decreased in the prefrontal cortex of HB-EGF KO mice (P,0.05 vs. control mice), despite comparable levels of total ERK expression (Fig. 6d). No significant differences in the expression of phosphorylated and total Akt or ...
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... significantly decreased in the prefrontal cortex of HB-EGF KO mice (P,0.05 vs. control mice) (Fig. 6c). Moreover, the expression of phosphorylated extracellular signal-regulated kinase (p-ERK) protein was significantly decreased in the prefrontal cortex of HB-EGF KO mice (P,0.05 vs. control mice), despite comparable levels of total ERK expression (Fig. 6d). No significant differences in the expression of phosphorylated and total Akt or ErbB4 proteins were observed between controls and HB-EGF KO mice (P.0.05 vs. control mice) ( Fig. 6e and f). On the other hand, we did not detect significant differences between controls and KO mice regarding the relative expression of other EGF family ...
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... (p-ERK) protein was significantly decreased in the prefrontal cortex of HB-EGF KO mice (P,0.05 vs. control mice), despite comparable levels of total ERK expression (Fig. 6d). No significant differences in the expression of phosphorylated and total Akt or ErbB4 proteins were observed between controls and HB-EGF KO mice (P.0.05 vs. control mice) ( Fig. 6e and f). On the other hand, we did not detect significant differences between controls and KO mice regarding the relative expression of other EGF family growth factors, such as EGF [Control; 10064.58 (mean6S.E.M., n = 10), KO; 10466.94 (n = 10)], TGF-a [Control; 100611.8 (n = 10), KO; 10469.36 (n = 10)], and betacelulin [Control; 10066.59 (n = ...
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Citations
... The same subunit was reported as a substrate for EGFR in glioma cells [119], where EGFR phosphorylates the COOH-terminal domain of the subunit, leading to an increase in glutamate-NMDAR signaling. Similarly, in an HB-EGF knockout mice model, the absence of the EGFR ligand led to reduced protein levels of the GluN1 subunit and calcium influx [120]. LTP, a maladaptive plasticity involved in numerous pathologies, including hyperalgesia, drug addiction, and tolerance, can be triggered by opioids in the spinal cord of rats and mice [100,101]. ...
... This leads to MOR lysosomal degradation via β-arrestin association, a subsequent impediment to interact with GPCR (comprised by α, β and γ subunits), and finally, hyperalgesia and tolerance. EGF knockout mice model, the absence of the EGFR ligand led to reduced protein levels of the GluN1 subunit and calcium influx [120]. LTP, a maladaptive plasticity involved in numerous pathologies, including hyperalgesia, drug addiction, and tolerance, can be triggered by opioids in the spinal cord of rats and mice [100,101]. ...
Oral cancer pain remains a significant public health concern. Despite the development of improved treatments, pain continues to be a debilitating clinical feature of the disease, leading to reduced oral mobility and diminished quality of life. Opioids are the gold standard treatment for moderate-to-severe oral cancer pain; however, chronic opioid administration leads to hyperalgesia, tolerance, and dependence. The aim of this review is to present accumulating evidence that epidermal growth factor receptor (EGFR) signaling, often dysregulated in cancer, is also an emerging signaling pathway critically involved in pain and opioid tolerance. We presented preclinical and clinical data to demonstrate how repurposing EGFR inhibitors typically used for cancer treatment could be an effective pharmacological strategy to treat oral cancer pain and to prevent or delay the development of opioid tolerance. We also propose that EGFR interaction with the µ-opioid receptor and glutamate N-methyl-D-aspartate receptor could be two novel downstream mechanisms contributing to pain and morphine tolerance. Most data presented here support that repurposing EGFR inhibitors as non-opioid analgesics in oral cancer pain is promising and warrants further research.
... However, other studies on ErbB4 knockout mice have provided controversial results: the signal blockade of neuregulin-1/ErbB4 elevates striatal dopamine levels [108,109]. This appears to be in accordance with the report that the gene ablation of HB-EGF, a ligand for ErbB1/B4 receptors, resulted in the abnormal behaviors relevant to the dopaminergic system or schizophrenia pathology [110]. Therefore, the neuregulin/ErbB4 functions in dopaminergic regulation differ significantly among target cell populations, their developmental stages, or neuregulin-splicing variants [65,111]. ...
Epidermal growth factor (EGF) and its homologs, such as neuregulins, bind to ErbB (Her) receptor kinases and regulate glial differentiation and dopaminergic/GABAergic maturation in the brain and are therefore implicated in schizophrenia neuropathology involving these cell abnormalities. In this review, we summarize the biological activities of the EGF family and its neuropathologic association with schizophrenia, mainly overviewing our previous model studies and the related articles. Transgenic mice as well as the rat/monkey models established by perinatal challenges of EGF or its homologs consistently exhibit various behavioral endophenotypes relevant to schizophrenia. In particular, post-pubertal elevation in baseline dopaminergic activity may illustrate the abnormal behaviors relevant to positive and negative symptoms as well as to the timing of this behavioral onset. With the given molecular interaction and transactivation of ErbB receptor kinases with Toll-like receptors (TLRs), EGF/ErbB signals are recruited by viral infection and inflammatory diseases such as COVID-19-mediated pneumonia and poxvirus-mediated fibroma and implicated in the immune–inflammatory hypothesis of schizophrenia. Finally, we also discuss the interaction of clozapine with ErbB receptor kinases as well as new antipsychotic development targeting these receptors.
... EGF knockout mice present behavioural abnormalities, including increased locomotor activity, damage to prepulse inhibition, and deficits in short-term memory. In addition, EGF knockout also reduces the activation of CaMKII and p21-activated kinase [35]. However, only a few studies focused on the function of EGF in patients with MDD, a previous study found that the concentration of EGF in plasma was significantly lower in patients with MDD [41], while we found that the expression of EGF was significantly increased after treatment with oleuropein combined with BDNF overexpression, which may contribute to reducing the inflammatory response after LPS stimulation. ...
Major depressive disorder (MDD) is a heterogeneous disease, involving multiple mechanisms and factors, which commonly result in injury to the psychosocial function of the central nervous system, and even suicidality of patients. However, effective treatment for MDD is still lacking. Oleuropein is a newly discovered natural compound extracted from olive leaves, which has a strong antioxidative effect by reducing the production of reactive oxygen species (ROS). Oleuropein also reduces blood pressure in humans and experimental animals, and protects blood vessels. Brain-derived neurotrophic factor (BDNF) is a member of the neurotrophin family, which supports the function of the central nervous system. BDNF plays an important role in the development of the nervous system via the regulation of cellular differentiation, survival neurogenesis and synaptic plasticity; therefore, we hypothesized that overexpression of BDNF might contribute to the therapeutic effect of oleuropein. Here, we first demonstrated that oleuropein reverses depressive-like behaviour and restores the inflammatory response in a mouse lipopolysaccharide (LPS) model of MDD. We further established a cell model of BDNF overexpression and inhibition in SH-SY5Y cells, and found that the concentration of intercellular calcium was increased after treatment with oleuropein combined with BDNF overexpression, which may be mediated by the BDNF-TrkB-CaMKII signalling pathway. In addition, we observed that the expression of neurotrophic factors, including epidermal growth factor (EGF), neurotrophin-3 (NT-3), and neurotrophin-4 (NT-4), was increased, which may be mediated by inhibition of the RhoA-ROCK signalling pathway.
... Both of these receptors have previously been implicated in psychiatric disorders, including schizophrenia, mood disorders, and antidepressant response [23][24][25][26][27][28][29]. Furthermore, dysregulation of HBEGF has been implicated in depression [30], and mice in which HBEGF have been knocked out display behavioral phenotypes, which are responsive to antipsychotics as well as altered dopamine and serotonin levels in the brain [31]. In addition to its relationship with escitalopram response, we also found it to be differentially expressed in the brain in individuals with depression who died by suicide, suggesting that our findings in the blood may also indicate central effects. ...
Identifying biomarkers of antidepressant response may advance personalized treatment of major depressive disorder (MDD). We aimed to identify longitudinal changes in gene expression associated with response to antidepressants in a sample of MDD patients treated with escitalopram. Patients ( N = 153) from the CAN-BIND-1 cohort were treated for 8 weeks, and depressive symptoms were assessed using the Montgomery-Åsberg Depression Rating Scale at 0, 2, 4, 6, and 8 weeks. We identified three groups of patients according to response status: early responders (22.9%), later responders (32.0%), and nonresponders (45.1%). RNA sequencing was performed in blood obtained at weeks 0, 2, and 8. RNA expression was modeled using growth models, and differences in the longitudinal changes in expression according to response were investigated using multiple regression models. The expression of RNAs related to response was investigated in the brains of depressed individuals, as well as in neuronal cells in vitro. We identified four RNAs ( CERCAM, DARS-AS1, FAM228B, HBEGF ) whose change over time was independently associated with a response status. For all except HBEGF , responders showed higher expression over time, compared to nonresponders. While the change in all RNAs differentiated early responders from nonresponders, changes in DARS-AS1 and HBEGF also differentiated later responders from nonresponders. Additionally, HBEGF was downregulated in the brains of depressed individuals, and increased in response to escitalopram treatment in vitro. In conclusion, using longitudinal assessments of gene expression, we provide insights into biological processes involved in the intermediate stages of escitalopram response, highlighting several genes with potential utility as biomarkers of antidepressant response.
... The EGF protein levels in the prefrontal cortex, striatum, and serum are lower in patients with schizophrenia [29,30]. In addition, conditional knockout of ventral forebrain-specific HB-EGF resulted in schizophrenia-like phenotypes, which could be ameliorated by typical or atypical antipsychotic agents [31]. Neonate animals treated with EGF exhibited schizophrenia-like behavioral abnormalities after puberty, some of which were ameliorated by treatment with antipsychotic agents [32]. ...
The phenomenon wherein the signaling by a given receptor is regulated by a different class of receptors is termed transactivation or crosstalk. Crosstalk between receptor tyrosine kinases (RTKs) and G protein-coupled receptors (GPCRs) is highly diverse and has unique functional implications because of the distinct structural features of the receptors and the signaling pathways involved. The present study used the epidermal growth factor receptor (EGFR) and dopamine D3 receptor (D3R), which are both associated with schizophrenia, as the model system to study crosstalk between RTKs and GPCRs. Loss-of-function approaches were used to identify the cellular components involved in the tyrosine phosphorylation of G protein-coupled receptor kinase 2 (GRK2), which is responsible for EGFR-induced regulation of the functions of D3R. SRC proto-oncogene (Src, non-receptor tyrosine kinase), heterotrimeric G protein Gβγ subunit, and endocytosis of EGFR were involved in the tyrosine phosphorylation of GRK2. In response to EGF treatment, Src interacted with EGFR in a Gβγ-dependent manner, resulting in the endocytosis of EGFR. Internalized EGFR in the cytosol mediated Src/Gβγ-dependent tyrosine phosphorylation of GRK2. The binding of tyrosine-phosphorylated GRK2 to the T142 residue of D3R resulted in uncoupling from G proteins, endocytosis, and lysosomal downregulation. This study identified the molecular mechanisms involved in the EGFR-mediated regulation of the functions of D3R, which can be extended to the crosstalk between other RTKs and GPCRs.
... HBEGF is a EGF-like binding partner of EGFR and mice lacking Hb-egf in the ventral forebrain showed abnormalities in psychomotor behavior and neurotransmission which can be ameliorated by typical or atypical antipsychotics (Oyagi et al., 2009). ...
Background:
Bipolar disorder (BD) is a common and highly heritable disorder of mood. Genome-wide association studies (GWAS) have identified several independent susceptibility loci. In order to extract more biological information from GWAS data, multi-locus approaches represent powerful tools since they utilize knowledge about biological processes to integrate functional sets of genes at strongly to moderately associated loci.
Methods:
We conducted gene set enrichment analyses (GSEA) using 2.3 million single-nucleotide polymorphisms, 397 Reactome pathways and 24,025 patients with BD and controls. RNA expression of implicated individual genes and gene sets were examined in post-mortem brains across lifespan.
Results:
Two pathways showed a significant enrichment after correction for multiple comparisons in the GSEA: GRB2 events in ERBB2 signaling, for which 6 of 21 genes were BD associated (PFDR = 0.0377), and NCAM signaling for neurite out-growth, for which 11 out of 62 genes were BD associated (PFDR = 0.0451). Most pathway genes showed peaks of RNA co-expression during fetal development and infancy and mapped to neocortical areas and parts of the limbic system.
Limitations:
Pathway associations were technically reproduced by two methods, although they were not formally replicated in independent samples. Gene expression was explored in controls but not in patients.
Conclusions:
Pathway analysis in large GWAS data of BD and follow-up of gene expression patterns in healthy brains provide support for an involvement of neurodevelopmental processes in the etiology of this neuropsychiatric disease. Future studies are required to further evaluate the relevance of the implicated genes on pathway functioning and clinical aspects of BD.
... Y-maze test. The Y-maze test was performed as previously described with minor modification 28 . The apparatus consisted of three arms (10 cm long × 40 cm wide × 12 cm high) made of gray plastic. ...
... The social interaction test was conducted as previously described with minor modification 28 . Two mice of the same genotype that were bred in different cages were placed in a plastic cage (24.5 cm long × 17.5 cm wide × 12.5 cm high) and allowed to explore freely for 10 min. ...
VGF nerve growth factor inducible (VGF) is a neuropeptide induced by nerve growth factor and brain-derived neurotrophic factor. This peptide is involved in synaptic plasticity, neurogenesis, and neurite growth in the brain. Patients with depression and bipolar disorder have lower-than-normal levels of VGF, whereas patients with schizophrenia and other cohorts of patients with depression have higher-than-normal levels. VGF knockout mice display behavioral abnormalities such as higher depressive behavior and memory dysfunction. However, it is unclear whether upregulation of VGF affects brain function. In the present study, we generated mice that overexpress VGF and investigated several behavioral phenotypes and the brain structure. These adult VGF-overexpressing mice showed (a) hyperactivity, working memory impairment, a higher depressive state, and lower sociality compared with wild-type mice; (b) lower brain weight without a change in body weight; (c) increased lateral ventricle volume compared with wild-type mice; and (d) striatal morphological defects. These results suggest that VGF may modulate a variety of behaviors and brain development. This transgenic mouse line may provide a useful model for research on mental illnesses.
... Unexpectedly, the HB-EGF blocking antibodies caused neurological side effects, which caused discontinuation of human trial [70]. Similar neurological effect was also shown in mice with brainspecific HB-EGF knockout [74]. Because ASO does not pass through blood brain barrier, we expect absence or far less brain-associated side effects in applying the ASO for human system [46]. ...
Objective
The upregulated expression of heparin binding EGF-like growth factor (HB-EGF) in the vessel and circulation is associated with risk of cardiovascular disease. In this study, we tested the effects of HB-EGF targeting using HB-EGF-specific antisense oligonucleotide (ASO) on the development of aortic aneurysm in a mouse aneurysm model.
Approach and results
Low-density lipoprotein receptor (LDLR) deficient mice (male, 16 weeks of age) were injected with control and HB-EGF ASOs for 10 weeks. To induce aneurysm, the mice were fed a high fat diet (22% fat, 0.2% cholesterol; w/w) at 5 week point of ASO administration and infused with angiotensin II (AngII, 1,000ng/kg/min) for the last 4 weeks of ASO administration. We confirmed that the HB-EGF ASO administration significantly downregulated HB-EGF expression in multiple tissues including the liver. Importantly, the HB-EGF ASO administration significantly suppressed development of aortic aneurysms including thoracic and abdominal types. Interestingly, the HB-EGF ASO administration induced a remarkable anti-hyperlipidemic effect by suppressing very low density lipoprotein (VLDL) level in the blood. Mechanistically, the HB-EGF targeting suppressed hepatic VLDL secretion rate without changing heparin-releasable plasma triglyceride (TG) hydrolytic activity or fecal neutral cholesterol excretion rate.
Conclusion
This result suggested that the HB-EGF targeting induced protection against aneurysm development through anti-hyperlipidemic effects. Suppression of hepatic VLDL production process appears to be a key mechanism for the anti-hyperlipidemic effects by the HB-EGF targeting.
... The Morris water maze test was performed as previously described (Oyagi et al., 2009(Oyagi et al., , 2012. A circular metal pool (diameter 120 cm  height 45 cm) was filled to a depth of 30 cm with water (20-22°C). ...
HYBID (HYaluronan Binding Protein Involved in hyaluronan [HA] Depolymerization, KIAA1199) is one of the HA binding proteins that is involved in the depolymerization of HA. HYBID mRNA is highly expressed in the brain, however, the role of HYBID in the brain remains unclear. In this study, we bred Hybid knock-out (KO) mice and evaluated the function of Hybid in the central nervous system. Hybid mRNA was expressed in the brain, especially in the hippocampus and cerebellum, in wild-type mice. Hybid KO mice demonstrated decreased mnemonic ability in novel object recognition and Morris water maze tests. The average molecular mass of hippocampal HA increased in KO mice, accompanied by a significant increase in the total HA amount. Hybid KO mice did not differ in behavior from wild-type mice in the open field test, evaluation of acoustic startle responses, or drug-induced seizure test. In real-time PCR, Hyal1 and Hyal2 mRNA levels, which code hyaluronidases 1 and 2, respectively, did not differ between the Hybid KO and wild-type mouse brain. These results indicate that Hybid plays a key role in memory function in the brain.
... MMPs are a group of calcium-dependent zinc-containing endopeptidases (Verma and Hansch, 2007). In terms of zinc and calcium, HB-EGF promotes cancerous cell proliferation via leukemia zinc -finger (PLZF) protein (Ozeki et al., 2013), and a calcium/calmodulin-cascade induces psychiatric disorders in HB-EGF knockout animals (Oyagi et al., 2009). Thus, it may be speculated that HB-EGF stimulates MMP9 and endopeptidase activities via zinc fingers, generating calcium signals that damage BBB in CCH. ...
Chronic cerebral hypoperfusion (CCH) manifests Alzheimer's Disease (AD) neuropathology, marked by increased amyloid beta (Aβ). Besides, hypoxia stimulates Heparin-binding EGF-like growth factor (HB-EGF) mRNA expression in the hippocampus. However, involvement of HB-EGF in CCH-induced Aβ pathology remains unidentified. Here, using Bilateral Common Carotid Artery Occlusion mouse model, we explored the mechanism of HB-EGF regulated Aβ induction in CCH. We found that HB-EGF inhibition suppressed, while exogenous-HB-EGF triggered hippocampal Aβ, proving HB-EGF-dependent Aβ increase. We also detected that HB-EGF affected the expression of primary Aβ transporters, receptor for advanced glycation end-products (RAGE) and lipoprotein receptor-related protein-1 (LRP-1), indicating impaired Aβ clearance across the blood-brain barrier (BBB). An HB-EGF-dependent loss in BBB integrity supported impaired Aβ clearance. The effect of HB-EGF on Amyloid Precursor Protein pathway was relatively insignificant, suggesting a lesser effect on Aβ generation. Delving into BBB disruption mechanism demonstrated HB-EGF-mediated stimulation of Matrix metalloprotease-9 (MMP9), which affected BBB via HB-EGF-ectodomain shedding and epidermal growth factor receptor activation. Examining the intersection of HB-EGF-regulated pathway and hypoxia revealed HB-EGF-dependent increase in transcription factor, Hypoxia-inducible factor-1alpha (HIF1α). Further, via binding to hypoxia-responsive elements in MMP9 gene, HIF1α stimulated MMP9 expression, and therefore appeared as a prominent intermediary in HB-EGF-induced BBB damage. Overall, our study reveals the essential role of HB-EGF in triggering CCH-mediated Aβ accumulation. The proposed mechanism involves an HB-EGF-dependent HIF1α increase, generating MMP9 that stimulates soluble-HB-EGF/EGFR-induced BBB disintegration. Consequently, CCH-mediated hippocampal RAGE and LRP-1 deregulation together with BBB damage impair Aβ transport and clearance where HB-EGF plays a pivotal role.
