Figure 1
Effect of HB-EGF on endogenous VEGF in vitro. (a) VEGF mRNA expression in mouse endothelial cell (CD31+ ) treated with 0, 10, and 100 ng/ml of recombinant human HB-EGF. (a') A phase-contrast micrograph displaying endothelial cell reaching approximately 70% confluent state prior to HB-EGF treatment. (b) VEGF mRNA expression in mouse non-endothelial cell (CD31− ) treated with 0, 10, and 100 ng/ml of recombinant human HB-EGF. Error bar represents the standard error (n = 4 for 1, 3, 6, and 12 h; n = 4 for 24 and 72 h data, respectively). (b') A phase-contrast micrograph displaying non-endothelial cell reaching
Source publication
Heparin binding epidermal growth factor-like growth factor (HB-EGF) is an angiogenic factor mediating radial migration of the developing forebrain, while vascular endothelial growth factor (VEGF) is known to influence rostral migratory stream in rodents. Cell migratory defects have been identified in animal models of hydrocephalus; however, the rel...
Contexts in source publication
Context 1
... In a higher dose (HB-EGF = 100 ng/ml), further increases (3.2-3.5 fold) in VEGF mRNA levels were detected at three time points (3,6 and 12 h) relative to the controls (p = 0.02, p = 0.01 and p = 0.03, respectively). In the negative controls (CD31 negative and α smooth muscle actin or α SMA positive), however, no significant change was observed (Fig. 1a,b). We then assayed the VEGF protein level in the conditioned media of CD31+ cell culture. The anti-CD31 immunoreactivity did not show a significant difference or an altered staining pattern between the vehicle control and HB-EGF (100 ng/ml) treated culture. VEGF protein was significantly elevated in the culture media of CD31+ endothelial ...
Context 2
... did not show a significant difference or an altered staining pattern between the vehicle control and HB-EGF (100 ng/ml) treated culture. VEGF protein was significantly elevated in the culture media of CD31+ endothelial cell treated with HB-EGF at 10 ng/ml, 100 ng/ml, and 1 μ g/ml as compared to the control (vehicle treated), respectively (Fig. 1c,d). This in vitro experiment has shown that exogenous administration of human soluble HB-EGF induces endoge- nous VEGF in the brain vascular endothelial ...
Context 3
... three groups of infusions were designed with coiled PE50 tube: Group 1 (n = 6) of SD rats received intraventricular VEGF-A 165 at 25 μ g/ml for 7 days and sacrificed on day 14. Group 2 (n = 5) of SD rats received intraventricular VEGF-A 165 at 25 μ g/ml for 7 days (Day 1 to 7) and intraventricular semaxanib (100 ng/ml) for 7 days (Day 8 to 14). Group 3 (n = 5) of SD rats received intraventricular VEGF-A 165 at 25 μ g/ml with bevacizumab at 25 mg/ml for 14 days. ...
Context 4
... of mice expressing human HB-EGF. A pIRES1hyg plasmid by Clontech (6061-1, Mountain View, CA) was used as the starting agent to create a vector for the HB-EGF overexpression transgene (Supplementary Fig. S1) B resistance for selection. The expression cassette contains human cytomegalovirus (CMV) major immediate early promoter/enhancer followed by a synthetic intron, multiple cloning site (MCS) known to enhance the sta- bility of the mRNA. ...
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Vascular endothelial growth factor (VEGF) is a potent growth factor playing diverse roles in vasculogenesis and angiogenesis. In the brain, VEGF mediates angiogenesis, neural migration and neuroprotection. As a permeability factor, excessive VEGF disrupts intracellular barriers, increases leakage of the choroid plexus endothelia, evokes edema, and...
Citations
... The interactional mechanism of the intracranial fluid and brain tissue is one of the controversial challenges in central nervous system disorders. Shedding light on the complexities of this mechanism can alleviate the pathophysiological ambiguities and biomechanical challenges in hydrocephalus [33][34][35][36] . The main goal of the present study is to introduce a mathematical framework to describe and dynamically compare the pulsatile interaction of blood, brain, and CSF in a hydrocephalus patient and a healthy subject based on their in vivo inputs. ...
Background
Shedding light on less-known aspects of intracranial fluid dynamics may be helpful to understand the hydrocephalus mechanism. The present study suggests a mathematical framework based on in vivo inputs to compare the dynamic interaction of pulsatile blood, brain, and cerebrospinal fluid (CSF) between the healthy subject and the hydrocephalus patient.
Method
The input data for the mathematical formulations was pulsatile blood velocity, which was measured using cine PC-MRI. Tube law was used to transfer the created deformation by blood pulsation in the vessel circumference to the brain domain. The pulsatile deformation of brain tissue with respect to time was calculated and considered to be inlet velocity in the CSF domain. The governing equations in all three domains were continuity, Navier-Stokes, and concentration. We used Darcy law with defined permeability and diffusivity values to define the material properties in the brain.
Results
We validated the preciseness of the CSF velocity and pressure through the mathematical formulations with cine PC-MRI velocity, experimental ICP, and FSI simulated velocity and pressure. We used the analysis of dimensionless numbers including Reynolds, Womersley, Hartmann, and Peclet to evaluate the characteristics of the intracranial fluid flow. In the mid-systole phase of a cardiac cycle, CSF velocity had the maximum value and CSF pressure had the minimum value. The maximum and amplitude of CSF pressure, as well as CSF stroke volume, were calculated and compared between the healthy subject and the hydrocephalus patient.
Conclusion
The present in vivo-based mathematical framework has the potential to gain insight into the less-known points in the physiological function of intracranial fluid dynamics and the hydrocephalus mechanism.
... Complex biomechanical recovery behavior of the hydrocephalic brain after shunting may an important reason for unpredictable complexities in shunt outcomes [57][58][59][60] . Hence, we performed a non-invasive computer simulation to calculate the volumes of CSF and brain, ICP, and exerted force on the brain tissue of hydrocephalus patients over 15 months after shunting. ...
The unpredictable complexities in hydrocephalus shunt outcomes may be related to the recovery behavior of brain tissue after shunting. The simulated cerebrospinal fluid (CSF) velocity and intracranial pressure (ICP) over 15 months after shunting were validated by experimental data. The mean strain and creep of the brain had notable changes after shunting and their trends were monotonic. The highest stiffness of the hydrocephalic brain was in the first consolidation phase (between pre-shunting to 1 month after shunting). The viscous component overcame and damped the input load in the third consolidation phase (after the fifteenth month) and changes in brain volume were stopped. The long-intracranial elastance (long-IE) changed oscillatory after shunting and there was not a linear relationship between long-IE and ICP. We showed the long-term effect of the viscous component on brain recovery behavior of hydrocephalic brain. The results shed light on the brain recovery mechanism after shunting and the mechanisms for shunt failure.
... The area calculated per each band was collected in the result file and saved in the spreadsheet. The relative fold change for each gene of interest was quantified with respect to the housekeeping gene (Schneider et al., 2012;Shim et al., 2013;Shim et al., 2016). ...
Symptoms of normal pressure hydrocephalus (NPH) and Alzheimer’s disease (AD) are somewhat similar, and it is common to misdiagnose these two conditions. Although there are fluid markers detectable in humans with NPH and AD, determining which biomarker is optimal in representing genetic characteristics consistent throughout species is poorly understood. Here, we hypothesize that NPH can be differentiated from AD with mRNA biomarkers of unvaried proximity to telomeres. We examined human caudate nucleus tissue samples for the expression of transient receptor potential cation channel subfamily V member 4 (TRPV4) and amyloid precursor protein (APP). Using the genome data viewer, we analyzed the mutability of TRPV4 and other genes in mice, rats, and humans through matching nucleotides of six genes of interest and one house keeping gene with two factors associated with high mutation rate: 1) proximity to telomeres or 2) high adenine and thymine (A + T) content. We found that TRPV4 and microtubule associated protein tau (MAPT) mRNA were elevated in NPH. In AD, mRNA expression of TRPV4 was unaltered unlike APP and other genes. In mice, rats, and humans, the nucleotide size of TRPV4 did not vary, while in other genes, the sizes were inconsistent. Proximity to telomeres in TRPV4 was <50 Mb across species. Our analyses reveal that TRPV4 gene size and mutability are conserved across three species, suggesting that TRPV4 can be a potential link in the pathophysiology of chronic hydrocephalus in aged humans (>65 years) and laboratory rodents at comparable ages.
... [53][54][55][56][57] We additionally identified a variant near HBEGF, a growth factor implicated in the pathobiology of CADASIL (Cerebral Autosomal Dominant Arteriopathy with Sub-cortical Infarcts and Leukoencephalopathy), 58 the major Mendelian prototype of vascular dementia. HBEGF also has an effect on angiogenesis, expression of VEGF-A, 59 on inflammation and oxidative stress and has been implicated in hydrocephalus. 60 APOE was strongly associated both with ACD and VaD in our meta-analysis. ...
Dementia is multifactorial with Alzheimer (AD) and vascular (VaD) pathologies making the largest contributions. Genome-wide association studies (GWAS) have identified over 70 genetic risk loci for AD but the genomic determinants of other dementias, including VaD remain understudied. We hypothesize that common forms of dementia will share genetic risk factors and conducted the largest GWAS to date of “all-cause dementia” (ACD) and examined the genetic overlap with VaD. Our dataset includes 809,299 individuals from European, African, Asian, and Hispanic ancestries with 46,902 and 8,702 cases of ACD and VaD, respectively. We replicated known AD loci at genome-wide significance for both ACD and VaD and conducted bioinformatic analyses to prioritize genes that are likely functionally relevant, and shared with closely related traits and risk factors. For ACD, novel loci identified were associated with energy transport ( SEMA4D ), neuronal excitability ( ANO3 ), amyloid deposition in the brain ( RBFOX1 ), and MRI markers of small vessel disease ( HBEGF ). Novel VaD loci were associated with hypertension, diabetes, and neuron maintenance ( SPRY2, FOXA2, AJAP1 , and PSMA3 ). Our study identified genetic risks underlying all-cause dementia, demonstrating overlap with neurodegenerative processes, vascular risk factors (Type-II diabetes, blood pressure, lipid) and cerebral small vessel disease. These novel insights could lead to new prevention and treatment strategies for all dementias.
... Western blot analysis was used to study the molecular mechanism of lysosomal accumulation caused by NAGLU knockdown in HUVEC. Previous studies have shown that extracellular HSPGs can bind and regulate the activity of HB-EGF (Iwamoto et al., 2010), whereas HB-EGF can induce VEGF production in several diseases (Nakai et al., 2009;Karakida et al., 2011;Shim et al., 2016). In addition, the VEGF-VEGFR2 pathway has been implicated in atherosclerosis (Khurana et al., 2005;Ylä-Herttuala et al., 2007;Taher et al., 2016). ...
... It has been proved that the extracellular HSPGs accumulation contributes to GAGs metabolism defects (Pan et al., 2005), which could bind and regulate the activity of HB-EGF (Iwamoto et al., 2010;De Pasquale et al., 2018). Even cells treated with HB-EGF in vitro could induce the production of VEGF (Nakai et al., 2009;Shim et al., 2016). In addition, a growing body of evidence suggests that activation of the VEGF-VEGFR2 pathway is involved in the GAGs metabolism (Zhang, 2010;Freudenberg et al., 2015) and the development of AS (Khurana et al., 2005;Ylä-Herttuala et al., 2007;Taher et al., 2016). ...
Cardiovascular diseases (CVDs), predominantly caused by atherosclerosis (AS), are the leading cause of mortality worldwide. Although a great number of previous studies have attempted to reveal the molecular mechanism of AS, the underlying mechanism has not been fully elucidated. The aberrant expression profiling of vascular endothelial cells (VECs) gene in early atherosclerosis (EAS) was analyzed according to the dataset (GSE132651) downloaded from the Gene Expression Omnibus (GEO) database. We primarily performed functional annotation analysis on the downregulated genes (DRGs). We further identified that α-N-acetylglucosaminidase (NAGLU), one of the DRGs, played a critical role in the progression of EAS. NAGLU is a key enzyme for the degradation of heparan sulfate (HS), and its deficiency could cause lysosomal accumulation and lead to dysfunctions of VECs. We found that siRNA knockdown of NAGLU in human umbilical vein endothelial cell (HUVEC) aggravated the abnormal accumulation of lysosomes and HS. In addition, the expression of NAGLU was reduced in the EAS model constructed by ApoE −/- mice. Furthermore, we also showed that heparin-binding EGF-like growth factor (HB-EGF) protein was upregulated while NAGLU knockdown in HUVEC could specifically bind to vascular endothelial growth factor receptor 2 (VEGFR2) and promote its phosphorylation, ultimately activating the phosphorylation levels of extracellular signal-regulated kinases (ERKs). However, the application of selective VEGFR2 and ERKs inhibitors, SU5614 and PD98059, respectively, could reverse the abnormal lysosomal storage caused by NAGLU knockdown. These results indicated that downregulation of NAGLU in HUVEC increases the abnormal accumulation of lysosomes and may be a potential biomarker for the diagnosis of EAS.
... The cause of this ventriculomegaly is claimed to be multi-factorial: excess CSF production, obstruction of CSF pathways, impaired CSF outflow, and/or imbalance of production and reabsorption at subarachnoid granulation (Ballabh, 2014;Bassan, 2009;Bateman, 2008;Bhattathiri et al., 2006;Garton et al., 2016;Kazan et al., 2005;Koschnitzky et al., 2018;Limbrick et al., 2010;Mbabazi-Kabachelor et al., 2019;McAllister, 2012;Murphy et al., 2002; that trajectories of doublecortin (DCX)-expressing young neurons in hydrocephalus were different in TMEM67 -/mutants as compared to WT during neonatal period (postnatal day 17, P17). A disorganized and widespread GFAP+ cells in the vicinity of radially and tangentially migrating neural progenitors in hydrocephalus is consistent with several previous studies by others (Doetsch and Alvarez-Buylla, 1996;Doetsch et al., 2002;Sawamoto et al., 2006;Shim et al., 2016) (Bothwell et al., 2019). In breeding animals, it has come to our attention that TMEM67 -/rats at P12 die in less than 10 days (~P21), even if many young neurons migrate postnatally in the presence of CH (Shim et al., 2019;Smith et al., 2006). ...
... Previously, we characterized three different models of hydrocephalus starting from human CSF assays (Shim et al., 2016;Shim et al., 2013;Shim et al., 2019). Hence, our genomic analysis was centered on factors associated with high mutation rates (Nusbaum et al., 2006) in hydrocephalus resulting from genetic mutations. ...
... Emmert et al., 2019;Shim et al., 2016;Shim et al., 2019) may shed light on mechanisms underlying post-hemorrhagic hydrocephalus. Similarly, finding a phenotype of "age-related loss or toxin-induced injury in neural connectivity(Luk et al., 2012)" along the nigrostriatal pathway might better explain how genetic and environmental factors alike can contribute to the pathogenesis of fPD. ...
Congenital hydrocephalus (CH) is caused by genetic mutations, but whether factors impacting human genetic mutations are disease-specific remains elusive. Given two factors associated with high mutation rates, we reviewed how many disease-susceptible genes match with (i) proximity to telomeres or (ii) high adenine and thymine (A + T) content in human CH as compared to other disorders of the central nervous system (CNS). We extracted genomic information using a genome data viewer. Importantly, 98 of 108 genes causing CH satisfied (i) or (ii), resulting in >90% matching rate. However, such a high accordance no longer sustained as we checked two factors in Alzheimer's disease (AD) and/or familial Parkinson's disease (fPD), resulting in 84% and 59% matching, respectively. A disease-specific matching of telomere proximity or high A + T content predicts causative genes of CH much better than neurodegenerative diseases and other CNS conditions, likely due to sufficient number of known causative genes (n = 108) and precise determination and classification of the genotype and phenotype. Our analysis suggests a need for identifying genetic basis of both factors before human clinical studies, to prioritize putative genes found in preclinical models into the likely (meeting at least one) and more likely candidate (meeting both), which predisposes human genes to mutations.
... A previous study showed that activation of miR-126 inhibited VEGFA expression, but the downstream pathway of VEGFA promoted neuronal survival [43,44]. In addition, increased expression of VEGFA has also been shown to have a dual regulatory role in neuronal survival [45,46]. SPRED1 affected the transmission of angiogenic signals and obstructed angiogenesis by inhibiting the MAP kinase pathway [15]. ...
miR-126 which is considered one of the most important miRNAs for maintaining vascular integrity, plays an important role in neuroprotection after cerebral ischemia–reperfusion (I-R). Moreover, vascular endothelial growth factor A (VEGFA), sprouty-related EVH1 domain-containing protein 1 (SPRED1), and Raf-1 are also involved in physiological processes of vascular endothelial cells (ECs). This study investigated how miR-126 changes with reperfusion time in different brain tissues after global cerebral ischemia and focal cerebral ischemia and examined the underlying mechanism miR-126 involving VEGFA, SPRED1, and Raf-1 after I-R. The results indicated decreases in the levels of miR-126-3p and miR-126-5p expression in mice and gerbils after I-R, consistent with the results after oxygen and glucose deprivation and reperfusion (OGD/R) in PC12 cells. Glial cells were activated as neuronal damage gradually increased after I-R. Inhibition of miR-126-3p exacerbated the OGD/R-induced cell death and reduced cell viability. After miR-126-3p inhibition, the levels of SPRED1 and VEGFA expression were increased, and p-Raf-1 expression was decreased after OGD/R. Moreover, based on the intervention of miR-126-3p inhibition, we found that the expression of p-Raf-1 was significantly increased after the intervention of siSPRED1, while it was not statistically significant after intervention of siVEGFA. The reduction of miR-126 expression after global and focal cerebral ischemia exacerbated neuronal death, which was closely related to increasing the SPRED1 activation and inhibiting the Raf-1 expression.
... The i.c.v. infusion of VEGF into the rat induced ventriculomegaly with an increase in SVZ neural progenitor cells in the rostral migratory stream, while the inhibition of VEGF signaling prevented these changes (Shim et al., 2016(Shim et al., , 2013. Thus, the present results suggest that VEGF-dependent endothelial proliferation and the vascular density of the CP Fig. 4. Effects of IVH on the proliferation of endothelial cells in the CP and ventricular area of the adult mouse brain. ...
The choroid plexus (CP), located at the walls of the brain ventricles, produces and secretes cerebrospinal fluid (CSF). Hydrocephalus is a neurological disorder in which the CP abnormally secretes excess amounts of CSF into the ventricles. There is currently no information on the vascular dynamics of the CP in adult brains under normal and hydrocephalic conditions. In the present study, we reported the continuous proliferation of endothelial cells in the CP of normal mice, which depended on vascular endothelial cell growth factor (VEGF). The proliferation of endothelial cells increased in mice with intraventricular hemorrhage, which was attenuated by a pretreatment with the toll-like receptor 4 (TLR4) inhibitor VIPER. Moreover, the intracerebroventricular infusion of the TLR4 agonist, lipopolysaccharide, increased endothelial cell proliferation in the CP and induced ventriculomegaly. The present results provide insights into the importance of the TLR4-initiated and VEGF-dependent proliferation of endothelial cells in the pathogenesis of hydrocephalus.
... In our work, we demonstrated that SS2 activated EGFR by increasing the shedding of the EGFR-associated ligands AREG, EREG, and HB-EGF, rather than interacting directly with EGFR. Previous studies show that AREG can upregulate mucin gene expression in obstructive airway diseases, EREG modulates Toll-like receptor (TLR)-mediated immune responses, and HB-EGF overexpression promotes vascular endothelial growth factor (VEGF) signaling resulting in hydrocephalus [56][57][58]. These ligands could induce the rapid phosphorylation of EGFR and subsequent intracellular signal transduction. ...
... These findings suggest a possibility that VEGF(R) is involved in the pathophysiologies not only in brain parenchyma but also in cerebral arteries after SAH. In addition, as EGFR-VEGFR cross-talk was demonstrated in cancer [13] and non-neoplastic diseases such as chronic hydrocephalus after SAH [14,15], we hypothesized that such a linkage might also contribute to post-SAH vasospasm development. Thus, this study was conducted to investigate whether EGFR activation is involved in post-SAH vasospasm, and if so, to reveal if the EGFR-VEGFR cross-talk contributes to signaling pathways downstream of EGFR in a mouse SAH model. ...
... Neurological impairments were blindly evaluated as previously described [18]. Briefly, neurological scores (3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18) were assessed by summing up six test scores (spontaneous activity; spontaneous movement of four limbs; forepaw outstretching; climbing; body proprioception; and response to whisker stimulation). Higher scores indicate better neurological status. ...
... Previous clinical trials targeting EGFR and VEGF in tumors suggested the importance to consider feed-back loops and cross-talk among different signaling pathways to obtain good results [13]. In addition, recent studies suggested that such cross-talk may also exist in non-neoplastic disease models: intraventricular infusion of recombinant human HB-EGF into rats as well as HB-EGF overexpression in mice caused hydrocephalus through activation of VEGF signaling [14,15]. Thus, it seems important issues to investigate whether such links exist in cerebral vasospasm after SAH, because the combination of EGFR and VEGFR inhibitors could be a strong therapeutic candidate for cerebral vasospasm. ...
Subarachnoid hemorrhage (SAH) is a devastating disease. Cerebral vasospasm is still an important cause of post-SAH poor outcomes, but its mechanisms remain unveiled. Activation of epidermal growth factor receptor (EGFR) is suggested to cause vasoconstriction in vitro, but no report has demonstrated the involvement of EGFR in vasospasm development after SAH in vivo. Cross-talk of EGFR and vascular endothelial growth factor (VEGF) receptor, which may affect post-SAH vasospasm, was also reported in cancer cells, but has not been demonstrated in post-SAH vasospasm. The aim of this study was to investigate whether EGFR as well as EGFR-VEGF receptor cross-talk engage in the development of cerebral vasospasm in a mouse SAH model. C57BL6 mice underwent endovascular perforation SAH or sham modeling. At 30 min post-modeling, mice were randomly administrated vehicle or 2 doses of selective EGFR inhibitors intracerebroventricularly. A higher dose of the inhibitor significantly prevented post-SAH neurological impairments at 72 h and vasospasm at 24 h associated with suppression of post-SAH activation of EGFR and extracellular signal-regulated kinase (ERK) 1/2 in the cerebral artery wall, especially in the smooth muscle cell layers. Anti-EGFR neutralizing antibody also showed similar effects. However, neither expression levels of VEGF nor activation levels of a major receptor of VEGF, VEGF receptor-2, were affected by SAH and two kinds of EGFR inactivation. Thus, this study first showed that EGFR-ERK1/2 pathways may be involved in post-SAH vasospasm development, and that EGFR-VEGF receptor cross-talk may not play a significant role in the development of vasospasm in mice.
