Search

Background: HOTAIR, one of the most widely studied long non-coding RNAs in tumors, is closely related to tumor proliferation, migration, invasion and chemoresistance. Objective: Here, we studied the mechanism behind proliferation and chemoresistance processes. Methods: A total of 75 samples were collected from patients who underwent surgical resection of their gastric cancer and received trastuzumab treatment. Primary cells were isolated and cultured. We also developed a cell line overexpressing HOTAIR by constructing a lentiviral vector. These cell lines were studied using an array of established biomolecular methods. Results: We found that HOTAIR levels were inversely associated with sensitivity to trastuzumab in gastric cancer and that overexpression of HOTAIR can promote the proliferation and invasion of gastric cancer cells. The sensitivity of cells overexpressing HOTAIR to two different types of human epidermal growth factor receptor 2 (HER2) inhibitors (trastuzumab and afatinib) showed that overexpression of HOTAIR is specific for trastuzumab resistance. Furthermore, luciferase reporter gene assay and western blot assay showed that there is a HOTAIR-miRNA330-ERBB4 competitive endogenous RNA regulatory network with miRNA330 as the core. Conclusion: HOTAIR can not only promote tumor proliferation but also enhance the resistance of tumor cells to drugs. Our experimental data not only showed strong expression of HOTAIR in gastric cancer, but also that strong expression of HOTAIR caused the sensitivity of gastric cancer cells to trastuzumab, which is a useful reference for postoperative medication.

The employment of an 8-hydroxyquinoline carbohydrazone tetradentate ligand, 8-hydroxyquinoline-2-carboxaldehyde-(benzoyl)hydrazone (H2L), HoCl3·6H2O and Et3N has led to the formation of dinuclear H...

Polycystic ovary syndrome (PCOS) is one of the most common endocrine disorders leading to infertility in women affecting reproductive, endocrine and metabolic systems. Recent genomewide association studies on PCOS cohorts revealed a single nucleotide polymorphism (SNP) in the ERBB4 receptor tyrosine kinase 4 gene, but its role in ovary development or during folliculogenesis remains poorly understood. Since no genetic animal models mimicking all PCOS reproductive features are available, we conditionally deleted Erbb4 in murine granulosa cells (GCs) under the control of Amh promoter. While we have demonstrated that Erbb4 deletion displayed aberrant ovarian function by affecting the reproductive function (asynchronous oestrous cycle leading to few ovulations and subfertility) and metabolic function (obesity), their ovaries also present severe structural and functional abnormalities (impaired oocyte development). Hormone analysis revealed an up-regulation of serum luteinizing hormone, hyperandrogenism, increased production of ovarian and circulating anti-Müllerian hormone. Our data implicate that Erbb4 deletion in GCs leads to defective intercellular junctions between the GCs and oocytes, causing changes in the expression of genes regulating the local microenvironment of the follicles. In vitro culture assays reducing the level of Erbb4 via shRNAs confirm that Erbb4 is essential for regulating Amh level. In conclusion, our results indicate a functional role for Erbb4 in the ovary, especially during folliculogenesis and its reduced expression plays an important role in reproductive pathophysiology, such as PCOS development.

The neuregulins (Nrgs 1–4) are a family of signaling molecules that play diverse roles in the nervous system. Nrg1 has been implicated in the formation of synapses and in synaptic plasticity. Previous studies have shown Nrg1 can affect neurite outgrowth in several neuronal populations, while the role of Nrg2 and Nrg3 in this process has remained understudied. The Nrgs can bind and activate the ErbB4 receptor tyrosine kinase which is preferentially expressed in GABAergic interneurons in the rodent hippocampus and cerebral cortex. In the present study, we evaluated the effects of Nrgs 1, 2, and 3 on neurite outgrowth of dissociated rat cortical ErbB4-positive (+)/GABA+ interneurons in vitro. All three Nrgs were able to promote neurite outgrowth during the first 2 days in vitro, with increases detected for both the axon (116–120%) and other neurites (100–120%). Increases in the average number of primary and secondary neurites were also observed. Treatment with the Nrgs for an additional 3 days promoted an increase in axonal length (86–96%), with only minimal effects on the remaining neurites (8–13%). ErbB4 expression persisted throughout the dendritic arbor and cell soma at all stages examined, while its expression in the axon was transient and declined with cell maturation. ErbB4 overexpression in GABAergic neurons promoted neurite outgrowth, an effect that was potentiated by Nrg treatment. These results show that Nrgs 1, 2, and 3 are each capable of influencing dendritic and axonal growth at early developmental stages in GABAergic neurons grown in vitro.

Background The regulatory roles of human epidermal growth factor receptor [erb-b2 receptor tyrosine kinase 4 (ERBB)] family in tumors was received widespread attention. Although ERBB4 was crucial regulator in metastasis of malignant tumors, the exact mechanism of ERBB4 in inflammatory breast cancer (IBC) remains unclarified. Methods In this study, we collected IBC tissues and cell lines, and explored the expression levels of ERBB4 and platelet-derived growth factor receptor alpha (PDGFRA) using real-time quantitative polymerase chain reaction (RT-PCR), immunohistochemistry (IHC) and western blot assays. Furthermore, cell viability with ERBB4 silencing in SUM149 cells was examined by MTT assay and cell migration and invasion were detected by transwell assay. Results The data indicated thatERBB4abolishing dramatically depressed capacity of proliferation, migration and invasion of IBC cells. Moreover, PDGFRA was an important factor for the function of ERBB4 and PDGFRA overexpression could, at least, partly rescue the ERBB4 silencing-mediated inhibition in proliferation and metastasis of IBC cells. Conclusions Take together, we verified the first time that ERBB4 promoted the progression of IBC through regulating PDGFRA. Thus, inhibition of ERBB4 might be a novel therapeutic candidate against IBC.

Background and purpose Amyotrophic lateral sclerosis (ALS)-related genes and mutations have been increasingly discovered recently and an improved understanding of genotype-phenotype relationships may help to predict the disease course and refine genetic diagnosis. Methods We collected clinical data and blood samples from 268 patients and used next-generation sequencing to comprehensively assay genetic variations in a panel of known ALS genes from 2015 to 2019. Results Among these patients, the mean age of onset was 52.30 ± 10.42 years with a mean diagnosis delay of 15.90 ± 11.88 months. Patients with SOD1, TARDBP and FUS variants were more likely to suffer from familial ALS. Additionally, carriers of FUS variants displayed the earliest onset, followed by those with SOD1 variants. Patients with NEFH variants showed a closer link to pesticide exposure. Patients with SETX variants were prone to bulbar onset with moderate anxiety problems. No genotype-phenotype relations were found in SPG11 and ERBB4 mutants. Conclusion Our findings uncovered some genotype-phenotype relationships and may help to predict the disease course of patients with ALS in southern China.

Background Amyotrophic lateral sclerosis/frontotemporal dementia (ALS/FTD) includes a large spectrum of neurodegenerative disorders. Objective To identify the relationship of ErbB4 mutation and ALS/FTD. Methods Here, we report an atypical case of frontal variant behavioral abnormalities at the initial stage, a stable plateau stage of 5 years, and paralysis involving both upper and lower motor neurons followed by progressive cognitive dysfunction at the advanced stage. The clinical findings suggested a diagnosis of ALS/FTD, and genetic testing revealed erb-b2 receptor tyrosine kinase 4 (ErbB4) heterozygous mutation (c.2136 T>G, p.I712M), identified in an ALS pedigree previously. We modeled mutant ErbB4 protein through the SWISS-MODEL Server, and speculated on the structural change caused by the mutation. We also identified that ErbB4 (I712M) mutation led to reduced auto-phosphorylation of ErbB4 upon neuregulin-1 (NRG1) stimulation. Results A functional analysis of ErbB4 mutation demonstrated an obviously decreased auto-phosphorylation of ErbB4 involving in the pathogenesis of ALS/FTD. Conclusion We firstly found ErbB4 mutation to be identified in ALS/FTD.

Amyotrophic lateral sclerosis (ALS) is a devastating motoneuron (Mn) disease without effective cure currently available. Death of MNs in ALS is preceded by failure of neuromuscular junctions and axonal retraction. Neuregulin 1 (NRG1) is a neurotrophic factor highly expressed in MNs and neuromuscular junctions that support axonal and neuromuscular development and maintenance. NRG1 and its ErbB receptors are involved in ALS. Reduced NRG1 expression has been found in ALS patients and in the ALS SOD1(G93A) mouse model; however, the expression of the isoforms of NRG1 and its receptors is still controversial. Due to the reduced levels of NRG1 type III (NRG1-III) in the spinal cord of ALS patients, we used gene therapy based on intrathecal administration of adeno-associated virus to overexpress NRG1-III in SOD1(G93A) mice. The mice were evaluated from 9 to 16 weeks of age by electrophysiology and rotarod tests. At 16 weeks, samples were harvested for histological and molecular analyses. Our results indicate that overexpression of NRG1-III is able to preserve neuromuscular function of the hindlimbs, improve locomotor performance, increase the number of surviving MNs, and reduce glial reactivity in the treated female SOD1(G93A) mice. Furthermore, the NRG1-III/ErbB4 axis appears to regulate MN excitability by modulating the chloride transporter KCC2 and reduces the expression of the MN vulnerability marker MMP-9. However, NRG1-III did not have a significant effect on male mice, indicating relevant sex differences. These findings indicate that increasing NRG1-III at the spinal cord is a promising approach for promoting MN protection and functional improvement in ALS. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s13311-019-00811-7) contains supplementary material, which is available to authorized users.

Kinase-targeted cancer therapy (KTCT) with ATP-competitive inhibitors has been widely applied in clinics. However, a number of kinase missense mutations were observed to confer acquired drug resistance during therapy, largely limiting the clinical application of kinase inhibitors in KTCT. Instead of directly influencing inhibitor binding, kinase mutations can also cause generic resistance to ATP-competitive inhibitors by increasing ATP affinity. Herein, the intermolecular interaction of the ATP molecule with clinically observed drug-resistant EGFR family kinase mutations involved in human cancer are systematically characterize. Rigorous quantum mechanics/molecular mechanics (QM/MM) calculation and empirical Poisson?Boltzmann/surface area (PB/SA) analysis as well as in vitro kinase assay and surface plasmon resonance analysis were integrated to explore the binding capability of ATP to mutant residues in the structural context of the kinase domain, which resulted in a comprehensive profile of ATP response to acquired drug-resistant mutations of four EGFR family kinases (EGFR/ErbB1, ErbB2, ErbB3 and ErbB4). From the profile, it was possible to identify those potent mutations that may influence ATP binding significantly; such mutations are potential candidates to cause generic resistance for ATP-competitive inhibitors. Consequently, the well documented generic drug-resistant mutation EGFR T790M and its counterpart ErbB2 T798M are found to increase ATP affinity by establishing an additional S?? interaction between the side-chain thioether group of the mutant Met residue and the aromatic adenine moiety of the ATP molecule, while EGFR D761Y is identified as a new generic drug-resistant mutation that can increase ATP affinity by eliminating unfavorable electrostatic repulsion. In contrast, ErbB2 K753E and T768I are considered to be two generic drug-sensitive mutations that can decrease ATP affinity by unfavorable charge reversal and by impairing favorable polar interaction, respectively. In addition, the EGFR L858R mutation is located at the kinase activation loop and nearby the kinase active site, thus largely complicating the multiply dependent relationship of kinase, ATP and inhibitor, which therefore exhibits divergent effects on different tested inhibitors.

This study investigated the role of ErbB4, the schizophrenia susceptibility gene, in synaptogenesis and synaptic transmission and its association with the pathogenesis of schizophrenia. We found that there was no significant difference in the size of inhibitory synapsin Gephyrin clusters in the dendrites of gamma-aminobutyric acid (GABA) neurons in the cultured cortex of ErbB4 knockout (ErbB4−/−) mice and ErbB4+/+ mice. The density and area of PSD-95 clusters in GABA-labelled neurons were not significantly different from those in the control group (p > 0.05). Specific pIkBα labelling of the pyramidal neurons and the axon initiation segments showed that the number of neuronal dendrites and the inhibitory postsynaptic proteins Gephyrin in the ErbB4 knockout group were significantly lower than those in the control group, whereas the size of the Gephyrin cluster remained unchanged. The number and area of the VGAT clusters on the somaclonal and axonal surfaces of the pyramidal neurons were also similar. ErbB4 knockout influenced excitatory synapses in intermediate neuronal cells and inhibitory synapses in pyramidal neurons, however, did not affect the inhibitory synapses in intermediate neuronal cells and excitatory synapses in pyramidal neurons. The number and area of PSD-95 clusters on the neuronal surface were significantly lower than those of the 1NMPP1 group, but the number and area of Gephyrin clusters on the neuronal surface were not changed (p > 0.05). These findings demonstrated that ErbB4 gene had no effect on the inhibitory synapses in interneurons and the excitatory synapses in pyramidal neurons.

Sharp wave ripples (SW-Rs) in the hippocampus are synchronized bursts of hippocampal pyramidal neurons (PyNs), critical for spatial working memory. However, the molecular underpinnings of SW-Rs remain poorly understood. We show that SW-Rs in hippocampal slices from both male and female mice were suppressed by neuregulin 1 (NRG1), an epidermal growth factor whose expression is enhanced by neuronal activity. Pharmacological inhibition of ErbB4, a receptor tyrosine kinase for NRG1, increases SW-R occurrence rate in hippocampal slices. These results suggest an important role of NRG1-ErbB4 signaling in regulating SW-Rs. To further test this notion, we characterized SW-Rs in freely moving male mice, chemical genetic mutant mice, where ErbB4 can be specifically inhibited by the bulky inhibitor 1NMPP1. Remarkably, SW-R occurrence was increased by 1NMPP1. We found that 1NMPP1 increased the firing rate of PyN neurons, yet disrupted PyN neuron dynamics during SW-R events. Furthermore, 1NMPP1 increased SW-R occurrence during both nonrapid eye movement (NREM) sleep states and wake states with a greater impact on SW-Rs during wake states. In accord, spatial working memory was attenuated in male mice. Together these results indicate that dynamic activity of ErbB4 kinase is critical to SW-Rs and spatial working memory. This study reveals a novel regulatory mechanism of SW-Rs and a novel function of the NRG1-ErbB4 signaling.SIGNIFICANCE STATEMENTSharp wave ripples (SW-Rs) are a hippocampal event, important for memory functioning. Yet the molecular pathways that regulate SW-Rs remain unclear. Neuregulin 1 (NRG1), previously known to be increased in pyramidal neuron's (PyNs) in an activity dependent manner, signals to its receptor, ErbB4 kinase, that is in important regulator of GABAergic transmission and long-term potentiation in the hippocampus. Our findings demonstrate that SW-Rs are regulated by this signaling pathway in a dynamic manner. Not only so, we show that this signaling pathway is dynamically needed for spatial working memory. These data suggest a molecular signaling pathway, NRG1-ErbB4, that regulates an important network event of the hippocampus, SW-Rs, that underlies memory functioning.

[ES] En el presente trabajo se abordará la estructura y función de los circuitos neuronales que participan en los procesos de formación de la memoria, y como estos se ven alterados en modelos animales en enfermedades del neurodesarrollo. Específicamente, se evaluará los déficits en el rendimiento cognitivo en un modelo murino de trastorno del neurodesarrollo (donde el gen Erbb4 se encuentra deleccionado) mediante estudios de una tarea conductual en roedores denominada ¿condicionamiento contextual del miedo¿. Además, estudiaremos las base neurobiológica de los trastornos conductuales mediante el marcaje y trazado de circuitos neuronales que consiste en la combinación de genética de roedores, imagen de fluorescencia y análisis neuroanatómico . De esta forma, se obtuvieron imágenes de dos áreas del cerebro del ratón - hipocampo y corteza prefrontal - tanto en animales control como en mutantes en condiciones con diferentes niveles de rendimiento cognitivo. Nuestros hallazgos nos permitirán entender qué circuitos neuronales están alterados en áreas específicas del cerebro en modelos animales deficientes en Erbb4 que recapitulan alteraciones cognitivas similares a las encontradas en enfermedades como la esquizofrenia, [EN] In this proyect, we will study the structure and function of neural circuits involved in the different memory formation process; and how they are altered in neurodevelopmental disease animal models. Specifically, we will evaluate cognitive performance déficits in neurodevelopment disease murine model (with the gen Erbb4 knocked out) through the behaviour anaylisis commonly call “contextual fear conditioning). Furthermore, we will study the neurological basis of the behavioural disorders through labeling of neural circuits, which implies a combination of mices´ genetics, fluorescence imaging and neuroanatomical analysis. This way, images from differents regions of the mices´brains – hipoccampus and prefrontal cortex – were obtained, both in control and mutants for Erbb4, in conditions with differents levels of cognitive performance. Our findings will allow us to understand what neural circuits are altered in specific brain areas, in animal models with a deficiency in Erbb4, which engage similar cognitive disruptions to the ones found in diseases as schizophrenia.

Resumen Las neuregulinas (NRGs) son una familia de factores de crecimiento tipo epidérmico que actúan modulando diversas respuestas celulares a través de la interacción con los receptores ErbBs. Las NRG1 y NRG2 son las principales en el Sistema Nervioso Central (SNC) y su principal receptor en el cerebro es el ErbB4. La señalización NRG1/ErbB4 ha sido involucrada en procesos relevantes para la formación y mantenimiento del SNC como la migración de interneuronas, la mielinización, la neurotransmisión, la modulación de sinapsis, entre otras. Por otro lado, la NRG2 también se une y activa al receptor ErbB4, sin embargo, su señalización ha sido menos explorada. Existe evidencia de que alteraciones funcionales en estos módulos de señalización podrían provocar disfunciones en redes neuronales, de relevancia en el contexto de enfermedades neuropsiquiátricas. Por lo tanto, es de gran importancia contar con modelos in vitro para el estudio de su señalización, caracterizados en cuanto a la proporción de tipos de neuronas en las cuales las proteínas se expresan, y a la expresión de sus genes. En el presente trabajo se determinó una proporción aproximada de 85% de neuronas glutamatérgicas y 15% de interneuronas GABAérgicas en cultivos primarios de neuronas de hipocampo de ratones C57BL/6 preparados del día embrionario (E) 18, similar a lo que se ha reportado in vivo e in vitro. Además, se determinó la expresión de los genes Nrg1 tipo III, Nrg2 y ErbB4 para el día in vitro (DIV) 7, siendo la expresión de Nrg1 tipo III mayor que Nrg2, similar a lo reportado en el hipocampo in vivo. Esto sugiere que los cultivos pueden ser utilizados como modelos in vitro válidos para el estudio de la señalización de las NRGs. Abstract Neuregulins (NRGs) comprise a family of EGF-like growth factors that modulate diverse cellular responses by interacting with their receptors ErbBs. In the Central Nervous System (CNS) the principal members include NRG1 and NRG2, and their principal receptor is ErbB4. The NRG1/ ErbB4 signaling has been involved in relevant processes for the development and maintenance of the CNS such as interneuron migration, myelinization, neurotransmission, and synapse modulation. NRG2 can also bind to ErbB4 and activate it: however, the NRG2/ErbB4 signaling remains less studied. There is evidence that functional disturbances in these signaling modules could lead to neuronal networks dysfunction, of importance in the context of neuropsychiatric diseases. Therefore, a characterized in vitro model in terms of the neuronal population in which these proteins are expressed, and their gene expression is greatly important for the study of NRG/ErbB4 signaling. Here we determined an approximate proportion of 85% glutamatergic neurons and 15% GABAergic interneurons in hippocampal cultures, prepared from C57BL/6 mice in embryonic (E) day 18, similar to the proportion reported in vivo and in vitro. We also determined the expression of Nrg1 type III, Nrg2 and ErbB4 genes by the day in vitro (DIV) 7. Nrg1 type III had higher expression levels than Nrg2, similar to the reported in the hippocampus in vivo. Together, these results suggest that these cultures can be used as a valid model for the study of NRGs signaling in vitro.

Extensive evidence suggests a dysfunction of the glutamate NMDA receptor (NMDAR) in schizophrenia, a severe psychiatric disorder with putative early neurodevelopmental origins, but clinical onset mainly during late adolescence. On the other hand, pharmacological models using NMDAR antagonists and the clinical manifestation of anti-NMDAR encephalitis indicate that NMDAR blockade/hypofunction can trigger psychosis also at adult stages, without any early developmental dysfunction. Previous genetic models of NMDAR hypofunction restricted to parvalbumin-positive interneurons indicate the necessity of an early postnatal impairment to trigger schizophrenia-like abnormalities, whereas the cellular substrates of NMDAR-mediated psychosis at adolescent/adult stages are unknown. Neuregulin 1 (NRG1) and its receptor ErbB4 represent schizophrenia-associated susceptibility factors that closely interact with NMDAR. To determine the neuronal populations implicated in “late” NMDAR-driven psychosis, we analyzed the effect of the inducible ablation of NMDARs in ErbB4-expressing cells in mice during late adolescence using a pharmacogenetic approach. Interestingly, the tamoxifen-inducible NMDAR deletion during this late developmental stage did not induce behavioral alterations resembling depression, schizophrenia or anxiety. Our data indicate that post-adolescent NMDAR deletion, even in a wider cell population than parvalbumin-positive interneurons, is also not sufficient to generate behavioral abnormalities resembling psychiatric disorders. Other neuronal substrates that have to be revealed by future studies, may underlie post-adolescent NMDAR-driven psychosis.

Neurons use local protein synthesis as a mechanism to support their morphological complexity, which requires independent control across multiple subcellular compartments including individual synapses. However, to what extent local translation is differentially regulated at the level of specific synaptic connections remains largely unknown. Here, we identify a signaling pathway that regulates the local synthesis of proteins required for the formation of excitatory synapses on parvalbumin-expressing (PV+) interneurons in the mouse cerebral cortex. This process involves the regulation of the mTORC1 inhibitor Tsc2 by the receptor tyrosine kinase ErbB4, which enables the local control of mRNA translation in a cell type-specific and synapse-specific manner. Ribosome-associated mRNA profiling reveals a molecular program of synaptic proteins that regulates the formation of excitatory inputs on PV+ interneurons downstream of ErbB4 signaling. Our work demonstrates that local protein translation is regulated at the level of specific connections to control synapse formation in the nervous system.

Background Immune checkpoint inhibitors (ICIs) have witnessed the achievements of convincing clinical benefits that feature the significantly prolonged overall survival (OS) of patients suffering from advanced non-small cell lung cancer (NSCLC), according to reports recently. Sensitivity to immunotherapy is related to several biomarkers, such as PD-L1 expression, TMB level, MSI-H and MMR. However, a further investigation into the novel biomarkers of the prognosis on ICIs treatment is required. In addition, there is an urgent demand for the establishment of a systematic hazard model to assess the efficacy of ICIs therapy for advanced NSCLC patients. Methods In this study, the gene mutation and clinical data of NSCLC patients was obtained from the TCGA database, followed by the analysis of the detailed clinical information and mutational data relating to two advanced NSCLC cohorts receiving the ICIs treatment from the cBioPortal of Cancer Genomics. The Kaplan–Meier plot method was used to perform survival analyses, while selected variables were adopted to develop a systematic nomogram. The prognostic significance of ERBB4 in pan-cancer was analyzed by another cohort from the cBioPortal of Cancer Genomics. Results The mutation frequencies of TP53 and ERBB4 were 54% and 8% in NSCLC, respectively. The mutual exclusive analysis in cBioPortal has indicated that ERBB4 does show co-occurencing mutations with TP53. Patients with ERBB4 mutations were confirmed to have better prognosis for ICIs treatment, compared to those seeing ERBB4 wild type (PFS: exact p = 0.017; OS: exact p p = 0.021). The mutation status of ERBB4 and TP53 was tightly linked to DCB of ICIs treatment, PD-L1 expression, TMB value, and TIICs. Finally, a novel nomogram was built to evaluate the efficacy of ICIs therapy. Conclusion ERBB4 mutations could serve as a predictive biomarker for the prognosis of ICIs treatment. The systematic nomogram was proven to have the great potential for evaluating the efficacy of ICIs therapy for advanced NSCLC patients.

As key inflammatory biomarkers C-reactive protein (CRP) and interleukin-6 (IL6) play an important role in the pathogenesis of non-inflammatory diseases, including specific cancers, such as breast cancer (BC). Previous genome-wide association studies (GWASs) have neither explained the large proportion of genetic heritability nor provided comprehensive understanding of the underlying regulatory mechanisms. We adopted an integrative genomic network approach by incorporating our previous GWAS data for CRP and IL6 with multi-omics datasets, such as whole-blood expression quantitative loci, molecular biologic pathways, and gene regulatory networks to capture the full range of genetic functionalities associated with CRP/IL6 and tissue-specific key drivers (KDs) in gene subnetworks. We applied another systematic genomics approach for BC development to detect shared gene sets in enriched subnetworks across BC and CRP/IL6. We detected the topmost significant common pathways across CRP/IL6 (e.g., immune regulatory, chemokines and their receptors, interferon γ, JAK-STAT, and ERBB4 signaling), several of which overlapped with BC pathways. Further, in gene–gene interaction networks enriched by those topmost pathways, we identified KDs—both well-established (e.g., JAK1/2/3, STAT3) and novel (e.g., CXCR3, CD3D, CD3G, STAT6)—in a tissue-specific manner, for mechanisms shared in regulating CRP/IL6 and BC risk. Our study may provide robust, comprehensive insights into the mechanisms of CRP/IL6 regulation and highlight potential novel genetic targets as preventive and therapeutic strategies for associated disorders, such as BC.

By knocking out all four EGFR ligands expressed in MDCK cells, this study shows the redundant and specific roles of each EGFR ligand in the ERK activation waves during collective cell migration., Epidermal growth factor receptor (EGFR) plays a pivotal role in collective cell migration by mediating cell-to-cell propagation of extracellular signal-regulated kinase (ERK) activation. Here, we aimed to determine which EGFR ligands mediate the ERK activation waves. We found that epidermal growth factor (EGF)–deficient cells exhibited lower basal ERK activity than the cells deficient in heparin-binding EGF (HBEGF), transforming growth factor alpha (TGFα) or epiregulin (EREG), but all cell lines deficient in a single EGFR ligand retained the ERK activation waves. Surprisingly, ERK activation waves were markedly suppressed, albeit incompletely, only when all four EGFR ligands were knocked out. Re-expression of the EGFR ligands revealed that all but HBEGF could restore the ERK activation waves. Aiming at complete elimination of the ERK activation waves, we further attempted to knockout NRG1, a ligand for ErbB3 and ErbB4, and found that NRG1-deficiency induced growth arrest in the absence of all four EGFR ligand genes. Collectively, these results showed that EGFR ligands exhibit remarkable redundancy in the propagation of ERK activation waves during collective cell migration.

Epidermal growth factor receptor (EGFR) is a vital intermediate in cell signaling pathway including cell proliferation, angiogenesis, apoptosis, and metastatic spread and also having four divergent members with similar structural features, such as EGFR (HER1/ErbB1), ErbB2 (HER2/neu), ErbB3 (HER3), and ErbB4 (HER4). Despite this, clinically exploited inhibitors of EGFR (including erlotinib, lapatinib, gefitinib, selumetinib, etc.) are not specific thus provoking unenviable adverse effects. Some of the paramount obstacles to generate and develop new lead molecules of EGFR inhibitors are drug resistance, mutation, and also selectivity which inspire medicinal chemists to generate novel chemotypes. The discovery of therapeutic agents that inhibit the precise stage in tumorous cells such as EGFR is one of the chief successful targets in many cancer therapies, including lung and breast cancers. This review aims to compile the various recent progressions (2016-2021) in the discovery and development of diverse epidermal growth factor receptor (EGFR) inhibitors belonging to distinct structural classes like pyrazoline, pyrazole, imidazole, pyrimidine, coumarin, benzothiazole, etc. We have summarized preclinical and clinical data, structure-activity relationships (SAR) containing mechanistic and in silico studies to provide proposals for the design and invention of new EGFR inhibitors with therapeutic significance. The detailed progress of the work in the field will provide inexorable scope for the development of novel drug candidates with greater selectivity and efficacy.

Diabetes mellitus is a major debilitating disease whose global incidence is progressively increasing with currently over 463 million adult sufferers and this figure will likely reach over 700 million by the year 2045. It is the complications of diabetes such as cardiovascular, renal, neuronal and ocular dysfunction that lead to increased patient morbidity and mortality. Of these, cardiovascular complications that can result in stroke and cardiomyopathies are 2- to 5-fold more likely in diabetes but the underlying mechanisms involved in their development are not fully understood. Emerging research suggests that members of the Epidermal Growth Factor Receptor (EGFR/ErbB/HER) family of tyrosine kinases can have a dual role in that they are beneficially required for normal development and physiological functioning of the cardiovascular system (CVS) as well as in salvage pathways following acute cardiac ischemia/reperfusion injury but their chronic dysregulation may also be intricately involved in mediating diabetes-induced cardiovascular pathologies. Here we review the evidence for EGFR/ErbB/HER receptors in mediating these dual roles in the CVS and also discuss their potential interplay with the Renin-Angiotensin-Aldosterone System heptapeptide, Angiotensin-(1-7), as well the arachidonic acid metabolite, 20-HETE (20-hydroxy-5, 8, 11, 14-eicosatetraenoic acid). A greater understanding of the multi-faceted roles of EGFR/ErbB/HER family of tyrosine kinases and their interplay with other key modulators of cardiovascular function could facilitate the development of novel therapeutic strategies for treating diabetes-induced cardiovascular complications.

Identification of candidate risk genes and alteration in the expression of proteins involved in regulating inhibitory neuron function in various psychiatric disorders, support the notion that GABAergic neuron dysfunction plays an important role in disease etiology. Genetic variations in neuregulin and its receptor kinase ErbB4, expressed exclusively by GABAergic neurons in the CNS, have been linked with schizophrenia. In the amygdala, ErbB4 is highly expressed in GABAergic intercalated cell clusters (ITCs), which play a critical role in amygdala-dependent behaviors. It is however unknown whether ErbB4 deletion from ITCs affects their synaptic properties and function in amygdala circuitry. Here, we examined the impact of ErbB4 deletion on inhibitory and excitatory circuits recruiting medial paracapsular ITCs (mpITCs) using electrophysiological techniques. Ablation of ErbB4 in mpITCs suppressed NMDA receptor-mediated synaptic transmission at thalamo-mpITC synapses and enhanced thalamic driven GABAergic transmission onto mpITCs. Furthermore, long-term potentiation (LTP) at thalamo-mpITC synapses was compromised in ErbB4 mutant mice, indicating that ErbB4 activity is critical for LTP at these synapses. Together, our findings suggest that ErbB4 deletion from mpITCs disrupts excitation-inhibition balance and learning mechanisms in amygdala circuits.

Le cancer de la prostate est le cancer le plus fréquemment diagnostiqué et la troisième cause de mortalité liée au cancer chez les hommes au Canada. Un quart des patients diagnostiqués développeront une forme plus agressive de ce cancer. Bien que nous possédions plusieurs indices cliniques pronostiques dans les cancers localisés (score de Gleason, taux sérique d’antigène prostatique spécifique (APS), stade, etc.), ceux-ci sont insuffisants pour adéquatement distinguer les patients à faible risque de progression de ceux à haut risque. A ce jour, aucun biomarqueur pronostique n’est encore utilisé en clinique. Les cliniciens ont donc besoin de nouveaux outils plus efficaces pour stratifier ce cancer et pour s’assurer d’adapter au mieux le traitement à chaque patient. En nous basant sur la littérature et sur des études préliminaires (cohortes de moins de 65 patients), notre hypothèse est que les protéines PUMA-NOXA et les récepteurs membranaires de la famille ERBB seraient, lorsqu’utilisés en combinaison, des biomarqueurs prédictifs de la progression du cancer de la prostate. Les objectifs de cette thèse sont : 1) identifier et valider de bons anticorps pour chaque biomarqueur d’intérêt, 2) définir les niveaux d’expression de chaque biomarqueur sur une cohorte de 285 patients, et 3) établir les corrélations entre les niveaux d’expression et les données cliniques des patients. Dans l’optique d’une utilisation en clinique, des anticorps de type monoclonal ont été choisis pour identifier les biomarqueurs d’intérêts. Ces anticorps ont été testés et validés pour leur spécificité par immunobuvardage de type western blot et par immunofluorescence. La localisation de la protéine d’intérêt a été validée sur des échantillons de tissus de patients suivie de l’optimisation du multi-marquage sur les cellules épithéliales et basales. Après perfectionnement de l’analyse d’images, nous avons montré qu’une expression extrême (faible ou forte) de PUMA couplée à une forte expression de NOXA dans les glandes bénignes est associée à la rechute biochimique des patients. La présence de ces biomarqueurs dans les glandes bénignes permet d’envisager d’améliorer l’identification lors des premières biopsies des patients se qualifiant pour la surveillance active. Par ailleurs, le suivi de l’expression des récepteurs de la famille ERBB dans les glandes tumorales permet une stratification des patients atteints d’un cancer de la prostate en fonction des risques de rechute biochimique, de développement de métastases et de mort liée au cancer. Ainsi, les patients présentant la combinaison d’une forte expression de EGFR et d’une faible expression de ERBB3 sont les plus susceptibles de mourir spécifiquement de leur cancer de la prostate, en particulier si les cellules tumorales présentes en plus une faible expression de ERBB2 entrainant un fort risque de développer des métastases. Mon projet de doctorat aura donc permis d’identifier et de valider des biomarqueurs d’intérêt pour prédire l’évolution du cancer de la prostate et démontrer l’intérêt de suivre ces biomarqueurs en combinaison afin d’obtenir une meilleure stratification des patients. Ces résultats devront être validés sur une cohorte indépendante et multicentrique en vue de fournir aux cliniciens un plus grand nombre d’outils pour leur permettre de réaliser une stratification fine des patients atteints d’un cancer de la prostate, et ouvrirait la voie à de nouvelles stratégies thérapeutiques plus ciblées., Prostate cancer is the most frequently diagnosed cancer and the third leading cause of cancer-related death in men in Canada. A quarter of patients will develop a more aggressive form of this cancer. While there are several clinical prognostic variables for localized prostate cancer (Gleason score, prostate specific antigen (PSA) levels, stage, etc.), these are insufficient to adequately distinguish between low and high-risk of progression cases. As a result, clinicians need new, more effective tools to stratify this cancer and to ensure that treatments are best tailored to each patient. To date, no prognostic biomarker has yet been used clinically. Based on the literature and preliminary studies of small cohorts (less than 65 patients), we hypothesize that the protein expression of PUMA-NOXA and ERBB family members could help with the prediction of prostate cancer progression. The objectives of this thesis are: 1) to identify and validate antibodies for each biomarker of interest, 2) to define the expression levels of each biomarker on a 285 patient cohort, 3) to evaluate the correlation between marker expression levels and patient clinical data. For clinical use, monoclonal-type antibodies were chosen to identify the biomarkers of interest. These antibodies were validated for specificity by western blot and immunofluorescence techniques. The localization of the protein of interest was further identified within samples of patient tissues and additional optimization involving combinatorial staining for epithelial and basal cells. After refining the imaging and statistical analysis of PUMA and NOXA in benign glands, we found that extreme (weak or strong) PUMA expression coupled with high NOXA expression was associated with biochemical relapse. In addition, these proteins have significant potential for predicting disease evolution based on the initial radical prostatectomy sample. The presence of these proteins in benign glands would allow the identification of patients less suitable for active surveillance. Additionally, statistical analysis of ERBB family receptors in tumor glands, when used alone, allow stratification of prostate cancer patients for the prediction of biochemical relapse, development of metastases and also specific death from prostate cancer. Moreover, patients expressing a combination of high EGFR expression and low ERBB3 expression are at high risk of biochemical relapse and are at higher risk of prostate cancer specific mortality. In addition, coupling this high EGFR – low ERBB3 combination to a low ERBB2 expression helps classify patients at high risk of developing metastases. My doctoral research project will have made it possible to identify and validate biomarkers of interest for predicting the progression of prostate cancer and demonstrating the interest of combining these biomarkers in order to achieve better stratification of patients with prostate cancer. In the context of clinical utility, these results need to be validated on an independent and multicenter cohort in order to confirm these findings. This would eventually provide clinicians with a greater number of tools at their disposal to correctly anticipate patient trajectories and possibly identify new therapeutic targets for the control of the disease.

A new class of pyrimidine derivatives were identified as potent protein tyrosine kinase (PTK) inhibitors for the treatment of idiopathic pulmonary fibrosis (IPF). Most of these small-molecule inhibitors displayed strong enzymatic activity against BTK and JAK3 kinases at concentrations lower than 10 nM. The representative compound N-(3-((5-chloro-2-(4-((1-morpholino)acetylamino)phenylamino)-4-pyrimidinyl)amino)phenyl)acrylamide (6 a) also exhibited high inhibitory potency toward both BTK and JAK kinase families, as well as ErbB4, at a concentration of 10 nM, achieving rates of inhibition higher than 57 %. Additionally, in vivo biological evaluations showed that 6 a can remarkably decrease the severity of IPF disease. All these investigations suggested that the multi-PTK inhibitor 6 a may serve as a promising agent for the treatment of IPF.

It is well established that the epidermal growth factor (EGF) receptor, receptor tyrosine-protein kinase erbB-2 (ERBB2)/human EGF receptor 2 (HER2), and, to a lesser extent, ERBB4/HER4, promote the pathogenesis of many types of human cancers. In contrast, the role that ERBB3/HER3, the fourth member of the ERBB family of receptor tyrosine kinases, plays in these diseases is poorly understood and, until recently, underappreciated. In large part, this was because early structural and functional studies suggested that ERBB3 had little, if any, intrinsic tyrosine kinase activity and, thus, was unlikely to be an important therapeutic target. Since then, however, numerous publications have demonstrated an important role for ERBB3 in carcinogenesis, metastasis, and acquired drug resistance. Furthermore, somatic ERBB3 mutations are frequently encountered in many types of human cancers. Dysregulation of ERBB3 trafficking as well as cooperation with other receptor tyrosine kinases further enhance ERBB3's role in tumorigenesis and drug resistance. As a result of these advances in our understanding of the structure and biochemistry of ERBB3, and a growing focus on the development of precision and combinatorial therapeutic regimens, ERBB3 is increasingly considered to be an important therapeutic target in human cancers. In this review, we discuss the unique structural and functional features of ERBB3 and how this information is being used to develop effective new therapeutic agents that target ERBB3 in human cancers.

HB-EGF is one of the most potent ligands of EGFR and ErbB4 receptors. This growth factor plays a pivotal role in many cellular processes, but its effect differs from one cell type to another and remains not fully understood. The aim of this work was to investigate the dependence between the rate of HB-EGF mediated cell proliferation and activation of EGFR and ErbB4 receptors. Therefore, the effects of human recombinant sHB-EGF (rsHB-EGF) on the proliferation of cell lines with different EGFR and ErbB4 quantity and ratio, as well as activation of the MARK-cascade p38 and ERK1/2 (p42/44) kinases, were analyzed. For comparison, a similar study of the effect of native sHB-EGF secreted by human histiocytic lymphoma cells U937 during co-cultivation with different cell lines was performed. It was proved that cell proliferation in response to sHB-EGF depends not only on the quantity but also on the ratio of EGFR and ErbB4. It was shown that signaling through ErbB4 is associated with activation of p38 kinase and signaling through EGFR associated with activation of ERK1/2 (p42/44) kinase. We assume the existence of two different mechanisms for sHB-EGF-mediated stimulation of cell proliferation, and the simultaneous launch of these mechanisms provides a maximal proliferative response. The results of this study support the feasibility of creating anti-proliferative drugs that target ErbB4.

Objective: This research was to establish a mitochondrial-related Drp1 gene and a lung cancer-related Erbb4 gene to participate in the regulatory network of lung cancer cell apoptosis, and to provide theoretical support for mitochondria to participate in tumor regulation. Method: The GO and KEGG methods were used to construct the regulatory networks of lung cancer related Drp1 and Erbb4 proteins that involved in the apoptosis of tumor cells, and to combine with the Bayesian network theory to screen out the largest possible action path acting on this network; The information about Drp1 in Oncomine database was collected, and the data in current database were analyzed twice. The role of Drp1 in lung cancer was meta-analyzed. Result: A regulatory network of Drp1 and Erbb4 involved in the apoptosis of tumor cells was successfully constructed; the optimal pathway was optimized using Bayesian theory; a total of 446 different types of research results were collected in the Oncomine database, of which there were 18 studies with statistical differences in Drp1 expression, 13 studies with increased Drp1’s expression, and 5 studies with decreased expression. Compared with the control group, Drp1 was expressed in lung cancer tissues highly (P