Epidermal Growth Factor Treatment Research Articles

The evaluation of melatonin and EGF interaction on breast cancer metastasis.

Metastasis in breast cancer is the first cause of death in patients. The epidermal growth factor (EGF) increases cancer cells' invasion, and migration. Melatonin's inhibitory effects on various types of cancer were confirmed. This study aimed to investigate whether melatonin could apply its impact through the EGF-related pathways or not. First, MDA-MB-231 and MCF7 cells were cultured, and then melatonin effects on cell viability were determined by MTT assay. Transwell invasion assay was applied to identify the invasiveness of these breast cancer cell lines under treatment of EGF and melatonin. Real-time RT-PCR then investigated the expression of MMP9 and MMP2 in determined groups. Cell proliferation was also assayed under EGF and melatonin treatment using Ki67 assessment by flow cytometry. The rate of invasion and migration of EGF-treated cells increased in both groups, in which melatonin caused increased invasion by EGF just in MCF7 cells. MMP9 and MMP2 expression increased significantly in both cell lines under EGF treatment, and melatonin increased these genes' expression in both cell lines (p<0.05). EGF increased the MMP9 and MMP2 gene expression, and melatonin increased EGF-induced expression (p<0.05). The EGF reduced the expression of the Ki67 protein in the MCF7 cell line, which was negatively affected by melatonin and EGF. In contrast, along with melatonin, EGF did not affect the proliferation of the MDA-MB-231 cell line. The results of this study show that melatonin in the presence of EGF does not show the anti-cancer properties previously described for this substance.

Natural products to enhance anti-tumor effects of trastuzumab and paclitaxel on the SKBR3, which is human breast cancer cell line.

e13051 Background: Neoadjuvant chemotherapy (ChT) with targeted therapy is the standard treatment for human epidermal growth factor 2 (HER2)- positive breast cancer (BC). In this context, Trastuzumab (Tz) and ChT significantly improved the pathological complete response (pCR) rate. Tz and Paclitaxel (PTX) combination therapy is also one of the neoadjuvant treatments used in HER2+BC, and despite the effectiveness of this treatment, there is a critical need for innovative approaches and further research that will improve the neoadjuvant response rate. Addressing this, the NaturaProDB tool presents a novel avenue for predicting in silico natural non-toxic compounds with potential anti-cancer properties. Our study aims to investigate synergistic effect of natural products proposed by this database on theurapeutic action of Tz and PTX on BC. Methods: The NaturaProDB database was searched using the ‘Target Cancer’ option in order to identify those natural products with anti-BC potential and the corresponding experimentally verified target genes. Using this database, Biochanin A (BCA) and Delphinidin (Dp) were selected as natural compounds because they both commonly inhibit the MAPK signaling pathway. We used the HER2+ SKBR3 cell line to determine IC20, IC30 and IC50 doses for BCA, Dp, Tz, and PTX through MTT assay. Notably, all experiments were done after 72 hrs treatment of all compounds. Further, various combinations within different concentrations of natural products and Tz and PTX were tested to define additive anti-proliferative effects on SKBR3 cell line. Results: We first defined two natural products with drug-like properties, BCA and Dp, which have anti-proliferative effect on BC. Treatment of SKBR3 with increasing concentrations of BCA and Dp caused dose-dependent proliferation inhibition and IC50 dose of compounds were defined as 42,70uM and 40,70uM, respectively. In addition, IC20 dose of Tz was 2,8µM, while IC30 dose of PTX was 2,3nM.Then, we showed that combinations of increasing doses of Dp (from 30µM to 90 µM) with IC20 Tz caused significant dose-dependent cell death compared to the control group (p&lt;0.0001). And the combinations of increasing doses of BCA (from 45µM to 360 µM) with IC20 Tz led similar significant results (p&lt;0.0001). Conclusions: Herein, it is shown that combining natural compounds (Dp, BCA) with conventional treatments (PTX,Tz) predicted for the SKBR3 cell line can enhance the inhibitory effect on cell proliferation. This study is one of the rare first cell line studies in which the synergistic effect of BCA and Dp, which are natural compounds selected using the NaturaProDB database, were combined with ChT and Tz to show their synergistic effect, and these findings indicate that synergistic combination treatments acting through different pathways (MAPK signaling pathway) can be used in the treatment of HER2+ and TN BC. It opens avenues for further research.

Regulation of STAT1 and STAT4 Expression by Growth Factor and Interferon Supplementation in Sjögren's Syndrome Cell Culture Models.

Our goal was to investigate the effects of epidermal growth factor (EGF) and interferons (IFNs) on signal transducer and activator of transcription STAT1 and STAT4 mRNA and active phosphorylated protein expression in Sjögren's syndrome cell culture models. iSGECs (immortalized salivary gland epithelial cells) and A253 cells were treated with EGF, IFN-alpha, -beta, -gamma, or mitogen-activated protein kinase p38 alpha (p38-MAPK) inhibitor for 0-24-48-72 h. STAT1 and STAT4 mRNA expression was quantified by qRT-PCR. Untreated and treated cells were compared using the delta-delta-CT method based on glyceraldehyde-3-phosphate dehydrogenase (GAPDH) normalized relative fold changes. phospho-tyrosine-701-STAT1 and phospho-serine-721-STAT4 were detected by Western blot analysis. STAT4 mRNA expression decreased 48 h after EGF treatment in A253 cells, immortalized salivary gland epithelial cells iSGECs nSS2 (sicca patient origin), and iSGECs pSS1 (anti-SSA negative Sjögren's Syndrome patient origin). EGF and p38-MAPK inhibitor decreased A253 STAT4 mRNA levels. EGF combined with IFN-gamma increased phospho-STAT4 and phospho-STAT1 after 72 h in all cell lines, suggesting additive effects for phospho-STAT4 and a major effect from IFN-gamma for phospho-STAT1. pSS1 and nSS2 cells responded differently to type I and type II interferons, confirming unique functional characteristics between iSGEC cell lines. EGF/Interferon related pathways might be targeted to regulate STAT1 and STAT4 expression in salivary gland epithelial cells. Further investigation is required learn how to better target the Janus kinases/signal transducer and activator of transcription proteins (JAK/STAT) pathway-mediated inflammatory response in Sjögren's syndrome.

Gastric Cancer: Clinical Features, Screening, Diagnosis, Treatment, and Prevention.

The objective of this article is to highlight the clinical features, screening, diagnosis, treatment, and prevention of gastric cancer (GC). Early GC is often asymptomatic leading to frequent delays in diagnosis. Weight loss and persistent abdominal pain are the most common symptoms at initial diagnosis. The diagnosis of GC typically involves a combination of endoscopy, biopsy, and imaging studies. Endoscopic resection techniques are emerging as successful treatment options for early GC. Treatment options for advanced GC include surgery and chemotherapy. The first line chemotherapy for advanced GC consists of doublet therapy with a combination of platinum and fluoropyrimidines. Trastuzumab, a monoclonal antibody, is used in the treatment of human epidermal growth factor 2 positive GCs. Antiangiogenic agents and immunotherapy are also useful in the treatment of GC. Currently there are no GC screening guidelines in the United States, but they exist in other regions where there is increased prevalence of GC. Prevention strategies for GC include Helicobacter pylori eradication and adoption of a healthy diet consisting of fruits and vegetables.

Artificial Dermis and Human Recombinant Epidermal Growth Factor Application for the Management of Critical Size Calvarial Defect.

Restoration of the 3-dimensional structure of the facial and calvarial skeleton after trauma or ablative oncologic surgeries serves as a framework for soft tissue reconstruction. In the present study, the authors aimed to evaluate the osteogenic effect of artificial dermis and epidermal growth factor treatment in critical-sized calvarial defects, which cannot be healed spontaneously. 8 mm calvarial defects were created in 28 male rats and filled with the artificial dermis, the artificial dermis and growth factor, growth factor or left untreated. Atomic absorption spectrometry was used to determine the amount of calcium, scanning electron microscopy was used to show the bone tissue in 3 dimensions, and immunohistochemistry was used to assess the bone formation and cell density. Histologic evaluation at 6 weeks showed incomplete bone regeneration in all groups. No statistical differences were found between the groups with regard to their scores for the following: inflammation, new bone formation, osteocyte density, resorption of bone at the edges of the defect, or fibrous tissue formation in the defect area. In conclusion, the predictability of bone formation in critical-size defects is not clear. Contrary to popular belief, the combined use of epidermal growth factor with artificial dermis or alone did not enhance the potential for osseous healing.

Real-world data on neoadjuvant chemotherapy with dual-anti HER2 therapy in HER2 positive breast cancer

BackgroundNeoadjuvant chemotherapy with dual-targeted therapy is the standard treatment for human epidermal growth factor 2 (HER2)-positive breast cancer. Although the dual-targeted therapy has significantly improved the pathological complete response (pCR) rate, further investigation is needed to identify biomarkers that predict the response to neoadjuvant therapy.MethodsThis retrospective study analyzed 353 patients with HER2-positive breast invasive ductal carcinoma. The correlation between clinicopathological factors and pCR rate was evaluated. A nomogram was constructed based on the results of the multivariate logistic regression analysis to predict the probability of pCR.ResultsThe breast pCR (b-pCR) rate was 56.1% (198/353) and the total pCR (t-pCR) rate was 52.7% (186/353). Multivariate analysis identified ER status, PR status, HER2 status, Ki-67 index, and neoadjuvant chemotherapy regimens as independent indicators for both b-pCR and t-pCR. The nomogram had an area under the receiver operating characteristic curve (AUC) of 0.73 (95% CI: 0.68–0.78). According to the nomogram, the t- pCR rate was highest in the ER-PR- HER2-positive patients (131/208) and lowest in the ER + PR + HER2-positive patients (19/73). The subgroup analyses showed that there was no significant difference in pCR rate among the neoadjuvant chemotherapy regimens in ER positive, PR positive, HER2 IHC 2 + , Ki67 index < 30% population. However, for ER-PR-HER2-positive patients, the neoadjuvant chemotherapy regimen has a great influence on the pCR rates.ConclusionsPatients with ER-negative, PR-negative, HER2 3 + and high KI-67 index were more likely to achieve pCR. THP may be used as an alternative to AC-THP or TCbHP in selected HER2-positive patients.

Tracheobronchial mucosal keratosis: A literature review of this rare disorder

BackgroundTracheobronchial mucosal keratosis (TBMK) is a rare airway disease that may cause refractory cough and airway stenosis. The characteristics of this disease remain unknown. In the present study, we describe this disorder based on a review of the current literature, emphasizing its diagnostic and therapeutic aspects. MethodsA comprehensive search of TBMK was performed in Medline, Google Scholar, Web of Science, Cochrane Library (UK), Embase, China National Knowledge Infrastructure (CNKI) (China), and Wan Fang Med Online (China). The following data were collected: patient characteristics, chest imaging findings, bronchoscopy, histopathologic findings, pathogen testing, treatment, and prognosis. ResultsAs of 2023, eighteen cases of TBMK have been reported. The main clinical manifestations were cough and expectoration. Chest imaging findings were non-specific. The main bronchoscopy findings were nodular protrusion of airway lumen and yellow-white purulent moss above the nodular lesion. The lesions were mainly located in the trachea and mainstem bronchus. The main pathological manifestations include keratinocytes or keratinocyte beads, squamous metaplasia, and mucosal inflammatory changes. The treatments that were administered include antibiotics, symptomatic treatment, and glucocorticoids. All methods were ineffective except for bronchoscopy-guided high-frequency electric knife and recombinant human epidermal growth factor treatment. ConclusionsTBMK is a rare respiratory disease with atypical clinical manifestations and chest computed tomography findings. Bronchoscopy revealed that nodular hyperplasia of the airway and purulent fur-covered lesions are typical manifestations. The final diagnosis needs to be confirmed by histopathological examination. There is a lack of effective treatment for this disease, and bronchoscopy-guided intervention therapy may be a candidate treatment.

Hyaluronic acid-based biodegradable microneedles loaded with epidermal growth factor for treatment of diabetic foot

Hyaluronic acid-based biodegradable microneedles loaded with epidermal growth factor for treatment of diabetic foot

Trim9 regulates the directional differentiation of retinal Müller cells to retinal ganglion cells.

Glaucoma is a leading cause of irreversible blindness, and effective therapies to reverse the visual system damage caused by glaucoma are still lacking. Recently, the stem cell therapy enable the repair and regeneration of the damaged retinal neurons, but challenges regarding the source of stem cells remain. This study aims to investigate a protocol that allows the dedifferentiation of Müller cells into retinal stem cells, following by directed differentiation into retinal ganglion cells with high efficiency, and to provide a new method of cellular acquisition for retinal stem cells. Epidermal cell growth factor and fibroblast growth factor 2 were used to induce the dedifferentiation of rat retinal Müller cells into retinal neural stem cells. Retinal stem cells derived from Müller cells were infected with a Trim9 overexpression lentiviral vector (PGC-FU-Trim9-GFP), and the efficiency of viral infection was assessed by fluorescence microscopy and flow cytometry. Retinoic acid and brain-derived neurotrophic factor treatments were used to induce the differentiation of the retinal stem cells into neurons and glial cells with or without the overexpression of Trim9. The expressions of each cellular marker (GLAST, GS, rhodopsin, PKC, HPC-1, Calbindin, Thy1.1, Brn-3b, Nestin, Pax6) were detected by immunofluorescence, PCR/real-time RT-PCR or Western blotting. Rat retinal Müller cells expressed neural stem cells markers (Nestin and Pax6) with the treatment of epidermal cell growth factor and fibroblast growth factor 2. The Thy1.1 positive cell rate of retinal stem cells overexpressing Trim9 was significantly increased, indicating their directional differentiation into retinal ganglion cells after treatment with retinoic acid and brain-derived neurotrophic factor. In this study, rat retinal Müller cells are dedifferentiated into retinal stem cells successfully, and Trim9 promotes the directional differentiation from retinal stem cells to retinal ganglion cells effectively.

Downregulation of cellular retinoic acid binding protein 1 fosters epithelial-mesenchymal transition in thyroid cancer.

Cellular retinoic acid binding protein 1 (CRABP1) participates in the regulation of retinoid signaling. Previous studies showed conflicting results regarding the role of CRABP1 in tumor biology, including protumorigenic and tumor-suppressive effects in different types of cancer. Our bioinformatics analyses suggested that CRABP1 expression was downregulated in thyroid cancer. Ectopic expression of CRABP1 in thyroid cancer cells suppressed migratory and invasive activity without affecting cell growth or cell cycle distribution. In transformed normal thyroid follicular epithelial cells, silencing of CRABP1 expression increased invasiveness. Additionally, CRABP1 overexpression was associated with downregulation of the mesenchymal phenotype. Kinase phosphorylation profiling indicated that CRABP1 overexpression was accompanied by a decrease in phosphorylation of epidermal growth factor (EGF) receptor and downstream phosphorylation of Akt, STAT3, and FAK, which were reversed by exogenous EGF treatment. Immunohistochemical analysis of our tissue microarrays revealed an inverse association between CRABP1 expression and disease stage of differentiated thyroid cancer. Taken together, our results suggest that CRABP1 expression is aberrantly lost in thyroid cancer, and this downregulation promotes the epithelial-mesenchymal transition at least partly through modulating EGF receptor signaling.

Effect of Growth Factors and Hormones during In Vitro Growth Culture of Cumulus-Oocyte-Complexes Derived from Small Antral Follicles in Pigs

This study evaluated the effect of various growth factors and hormones in an in vitro growth (IVG) medium on the in vitro maturation (IVM) and developmental competence of oocytes derived from small antral follicles (SAFs) in pigs. Cumulus-oocyte complexes (COCs) derived from SAFs were either untreated or treated with epidermal growth factor (EGF), insulin-like factor-1 (IGF-1), insulin, or growth hormone (GH) for 2 days of IVG. Following IVG, COCs were cultured for maturation, and IVM oocytes were induced for parthenogenesis (PA). During IVG, the nuclear maturation of oocytes was significantly increased by the insulin treatment compared to other treatments. Moreover, the insulin treatment significantly increased blastocyst formation after PA relative to the No-IVG, control, EGF, and GH treatments. The cumulus expansion score after IVG-IVM was significantly higher in the insulin group than in the other groups. The glutathione (GSH) contents in IVM oocytes were increased through treatment with IGF, insulin, and GH compared to those of No-IVG oocytes. The level of reactive oxygen species (ROS) in IVM oocytes in all treatment groups was significantly lower after IVG culture than in the No-IVG group. The maturation-promoting factor (MPF) activity after IVM in the insulin-treated oocytes was significantly higher than that of the oocytes treated with EGF, IGF-1, and GH. In conclusion, this study demonstrates that insulin treatment during IVG culture improves the maturational and developmental competence of oocytes derived from SAFs in pigs through its effect on cumulus cell expansion and cytoplasmic microenvironments, such as GSH, ROS, and MPF activity.

Abstract P4-07-35: Changes in Left Ventricular Global Longitudinal Strain in breast cancer patients receiving anti-HER2 and/or Adriamycin therapy and outcomes with early implementation of cardio-protective measures

Abstract Background: The treatment of Human Epidermal Growth Factor -2 (HER2) positive breast cancer has advanced since Trastuzumab and subsequently Pertuzumab were introduced and approved as antibody-targeted therapy. With the incorporation of anti-HER2 therapy, cardiotoxicity poses a significant risk and is a well know side effect. This toxicity can increase with concomitant use of Anthracyclines. Left Ventricular Global Longitudinal Strain is assessed using speckle tracking analysis on 2D echocardiogram and a relative reduction in LV global longitudinal strain (GLS) of 10-15% from baseline appears to have specificity to predict downstream reduction in Left Ventricular Ejection Fraction (LVEF). There is growing use for identification of GLS changes in these patients and its incorporation into medical decision making that impacts oncological and cardiac care. Methods: We conducted an analysis of 200 patients receiving anti-HER2 therapy at Henry Ford Cancer institute from Jan 1, 2016 to June 1, 2022 to determine if there was a 10-15% reduction in GLS detected prior to a decrease in ejection fraction and if and how these detections resulted in the implementation of cardio protective measures and downstream effects on cancer therapy. Results: There were 198 patients with GLS and LVEF data. 175 patients (88.3%) completed one year of cancer therapy 107/198 patients (54%) had no change in GLS or LVEF. 91/198 patients (45%) had changes in GLS and or LVEF. 41/91 (45%) patients with LVEF decline did not have cardioprotective intervention implemented. Despite this, 34 (83%) of these patients completed cancer treatment. 50/91 (55%) patients did have cardioprotective interventions with 40 (78%) patients from this subgroup completing cancer treatment. 81% of patients with EF decline were able to complete treatment. 50% of patients with EF decline had EF recovery within 6 months of completion of cancer therapy. The odds of LVEF decline were 9.8 times higher for those with GLS decline (OR=9.0, p&amp;lt; 0.001). Patients with LVEF decline were more likely to have cardio preventive intervention (OR=18.8, p&amp;lt; 0.001). Multivariate analysis did not find an association between cardiac risk factors such as hypertension, diabetes mellitus, smoking, obesity and hyperlipidemia with GLS decline. There was no disparity by race. Conclusion: Our study revealed that early changes in GLS and LVEF did not impact the completion of cancer treatment irrespective of implementation of cardioprotective measures. Citation Format: Hussna E. Abunafeesa, Cortney Mckay, Pin Li, Madhulata Reddy, Vrushali Dabak. Changes in Left Ventricular Global Longitudinal Strain in breast cancer patients receiving anti-HER2 and/or Adriamycin therapy and outcomes with early implementation of cardio-protective measures [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P4-07-35.

Red LED light therapy associated with epidermal growth factor on wound repair process in rats.

Epidermal growth factor (EGF) and light-emitting diode (LED) are currently deployed as promissory treatments for skin repair; however, the mechanisms of their association are not yet evidenced. Thus, the present study aimed to evaluate the effects of combined treatment with EGF and red LED on the wound healing processes in rats. Adult Wistar rats were randomized in control group (CG) wounds without treatment; wounds submitted to EGF treatment (EGF); wounds submitted to LED treatment (LED); wounds submitted to EGF associated with LED treatments (EGF/LED). Treatments were performed immediately after the surgical procedure and each 24h, totaling 8 sessions. Moreover, LED was applied before EGF treatment at a single point in the center of the wound. Morphological characteristics and the immunoexpression of COX-2, VEGF, and TGF-β were measured. The results demonstrated that EGF/LED group presented a higher wound healing index. Additionally, all experimental groups presented similar findings in the histological evaluation, the degree of inflammation, and the area of dermis-like tissue. However, for EGF-treated animals (with or without LED), neoepithelial length was higher. Furthermore, all the treated groups decreased COX-2 and increased VEGF immunoexpression, and only EGF/LED group enhanced the TGF-β protein expression when compared to the untreated group. This research shows that EGF and LED modulate inflammatory process and increase the vascularity. In addition, treatment of EGF associated with LED promoted a more evident positive effect for increasing TGF-β expression and may be promising resources in the clinical treatment of cutaneous wounds.

EGF promotes human trophoblast cell invasion by downregulating CTGF expression via PI3K/AKT signaling.

Although the pro-invasive role of epidermal growth factor (EGF) has been reported in human trophoblast cells, the underlying mechanism remains largely unexplored. This work reveals that EGF-induced downregulation of connective tissue growth factor (CTGF) mediates the EGF-stimulated human trophoblast cell invasion. During the development of the placenta, trophoblast cell invasion must be carefully regulated. Although EGF has been shown to promote trophoblast cell invasion, the underlying mechanism remains largely undetermined. Our previous study using RNA-sequencing (RNA-seq) has identified that kisspeptin-1 is a downstream target of EGF in a human trophoblast cell line, HTR-8/SVneo, and mediates EGF-stimulated cell invasion. In the present study, after re-analysis of our previous RNA-seq data, we found that the CTGF was also downregulated in response to the EGF treatment. The inhibitory effects of EGF on CTGF mRNA and protein levels were confirmed in HTR-8/SVneo cells by reverse transcription quantitative real-time PCR and western blot, respectively. Treatment with EGF activated both PI3K/AKT and ERK1/2 signaling pathways. Using pharmacological inhibitors, our results showed that EGFR-mediated activation of PI3K/AKT signaling was required for the EGF-downregulated CTGF mRNA and protein levels. Matrigel-coated transwell invasion assays demonstrated that EGF treatment stimulated cell invasion. In addition, the invasiveness of HTR-8/SVneo cells was suppressed by treatment with recombinant human CTGF. By contrast, siRNA-mediated knockdown of CTGF increased cell invasion. Notably, the EGF-promoted HTR-8/SVneo cell invasion was attenuated by co-treatment with CTGF. This study provides important insights into the molecular mechanisms mediating EGF-stimulated human trophoblast cell invasion and increases the understanding of the biological functions of CTGF in the human placenta.

Abundance of Dual Specificity Phosphatase (DUSP) 1 and DUSP6 mRNA Is Regulated by Hippo Signaling in Bovine Pre-ovulatory Granulosa Cells.

Ovulatory disorders are a major cause of infertility in humans as well as economically important species. In physiological conditions, the LH surge induces the expression of epidermal growth factor (EGF)-like ligands that activate the EGR receptor (EGFR) and subsequently the mitogen-activated protein kinase (MAPK) pathway. The magnitude and duration of MAPK phosphorylation are regulated by dual-specificity phosphatases (DUSPs). Besides this well-known cascade, other signaling pathways such as the Hippo pathway modulate the ovulatory cascade and are reported to crosstalk with MAPK signaling. Here, we tested the hypothesis that LH and the Hippo pathway regulate DUSP expression in bovine pre-ovulatory granulosa cells. The abundance of DUSP6 mRNA but not DUSP1 was decreased by LH (P < 0.05). Cells were then pre-treated (1h) with two inhibitors of Hippo signaling, verteporfin (1µM) or peptide-17 (25µM), before exposure for 6h to LH or to EGF. Treatment with verteporfin increased DUSP1 mRNA levels (P < 0.05) in the presence or absence of EGF or LH and treatment with peptide-17 increased DUSP6 and not DUSP1 mRNA abundance. These data indicate a differential regulation of DUSP1 and DUSP6 mRNA by the Hippo pathway in pre-ovulatory granulosa cells, which suggests a complex control of MAPK signaling around ovulation.

Human Epidermal Growth Factor Improves Atopic Disease-like Skin Lesions in DFE/DNCB Induced BALB/c Mice and Human Keratinocytes

PURPOSE: Atopic skin disease is an inflammatory disease with a multifactorial pathogenesis that presents as itchy, dry skin. Human epidermal growth factor (EGF) plays an essential role in regulating cell growth, proliferation, and differentiation. Accordingly, this study aimed to investigate the effects of EGF on atopic diseases.METHODS: Atopic disease was induced by the &lt;i&gt;Dermatophagoides farinae&lt;/i&gt; house dust mite extract and 2,4-dinitrochlorobenzene in BALB/c mice. The mice were divided into the following five atopic disease induction and treatment groups: control, atopic disease only, atopic disease+epidermal growth factor-1, atopic disease+epidermal growth factor-10, and positive control atopic disease+ceramide. We examined dermal and epidermal ear thickness, mast cell and T cell infiltration, serum immunoglobulin E and immunoglobulin G2a levels, and mRNA expression of pathogenic cytokines in murine ear tissue and human keratinocyte cells.RESULTS: Epidermal growth factor treatment increased epidermal and dermal ear thickness and inhibited mast and T cell infiltration in the ear in a dose-dependent manner. Treatment also suppressed immunoglobulin E and G2a levels and the mRNA expression of pathogenic cytokines in murine ear tissue and human keratinocytes. The therapeutic outcome of epidermal growth factor was stronger than that of the positive control ceramide.CONCLUSIONS: EGF may be beneficial in the treatment of atopic disease. Key words: Epidermal growth factor, Atopic disease, Keratinocyte cells, Cytokine

Norcantharidin attenuates epidermal growth factor-induced proliferation, EMT and motility in ARPE-19 cells by modulating the AKT/snail/E-cadherin axis

AimsNorcantharidin (NCTD) is a demethylated derivative of cantharidin demonstrated to have anti-proliferative, anti-inflammatory, and anti-fibrosis properties. The purpose of the current study is to investigate the underlying mechanisms and signaling pathways affected by NCTD in human ARPE-19 cells. Main methodsCell growth and rate of proliferation were assayed by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay, colony formation assay, and cell cycle distribution/quantification. Cell motility was detected with in vitro migration assay. The level of epithelial-mesenchymal transition (EMT)-related proteins and mRNA (Snail, Slug, E-cadherin) were detected using Western blotting, real-time quantitative polymerase chain reaction, and immunofluorescence assay. Overexpression of Snail plasmid was determined by transfection assay. Key findingsWe found that NCTD reduced epidermal growth factor (EGF)-induced ARPE-19 cell viability and proliferation through increasing the p21 and p27 expression and decreasing the cyclin D1 expression. NCTD also inhibited EGF-mediated EMT and cell motility through increased protein and mRNA levels of E-cadherin and decreased Snail in EGF-induced ARPE-19 cells. Overexpression of Snail significantly decreased ARPE-19 cell motility and increased E-cadherin expression in NCTD-treated cells. Additionally, when NCTD was combined with a PI3K inhibitor (LY294002) significantly decreased the p-AKT and Snail expression, and increased the E-cadherin expression of EGF treatment in ARPE-19 cells. SignificanceThe current findings revealed that NCTD suppresses the EGF-induced proliferation, motility, and EMT of ARPE-19 cells through inactivation of the AKT-mediated Snail/E-cadherin pathway. NCTD may be a potential preventive agent for proliferative vitreoretinopathy.

1001P Epidermal growth factor receptor (EGFR) testing and treatment patterns associated with diagnosis of non-small cell lung cancer (NSCLC) with EGFR exon 20 insertions (ex20ins) in the US

With new targeted therapies approved for patients with NSCLC with EGFRex20ins, it is important to understand real-world diagnosis and treatment choice, as these mutations are associated with poor prognosis and require tailored treatment. The Flatiron Health electronic health record database was used to identify 75,067 patients with advanced NSCLC from 01/01/2011 to 12/31/2021. EGFR testing rates and proportion of tests done with next-generation sequencing (NGS) were described for all patients. First-line (1L) treatments were described for patients with EGFRex20ins(+) results in the last 5 years (2017-2021), stratified by initiation of 1L before or after EGFRex20ins(+) testing result. Overall, 67% of patients were tested for EGFR mutations (n=51,470). EGFR testing rates and the proportion of tests done with NGS increased from 2011 to 2021 (44.4 to 80.7% and <1.0 to 81.0%, respectively). Among patients with EGFRex20ins(+) results 2011-2017 (n = 250), 59 (24%) started 1L treatment prior to and 141 (56%) started 1L after their first EGFRex20ins(+) (50 were unknown). A higher proportion of patients received EGFR TKIs or were enrolled in clinical trials, and a lower proportion received chemotherapy if they started 1L treatment after their first EGFRex20ins(+) result, when compared to patients who started 1L treatment before. Similar proportions of patients were treated with IO monotherapy or IO + chemotherapy regardless of when treatment was started (table).Table: 1001P1L treatments in patients with EFRex20ins 2017-2021Started treatment before EGFRex20ins (+) result n (%)Started treatment after EGFRex20ins (+) result n (%)n = 59n = 141Chemotherapy27 (45.8)36 (25.5)IO monotherapy5 (8.5)16 (11.3)IO + chemotherapy21 (35.6)44 (31.2)EGFR TKI4 (6.8)32 (22.7)Clinical Trial1 (1.7)12 (8.5)Other1 (1.7)1 (0.7) Open table in a new tab EGFR testing increased over time; however, 20% of patients still remained untested in 2021. Among patients with EGFRex20ins mutations, a high proportion of patients started IO or EGFR TKIs after genetic testing despite limited effectiveness in this population. With the recent approval of therapies targeting EGFRex20ins mutations, increased testing and awareness of treatment outcomes in this population are needed.

Cellular context alters EGF-induced ERK dynamics and reveals potential crosstalk with GDF-15.

Cellular signaling dynamics are sensitive to differences in ligand identity, levels, and temporal patterns. These signaling patterns are also impacted by the larger context that the cell experiences (i.e., stimuli such as media formulation or substrate stiffness that are constant in an experiment exploring a particular variable but may differ between independent experiments which explore that variable) although the reason for different dynamics is not always obvious. Here, we compared extracellular-regulated kinase (ERK) signaling in response to epidermal growth factor treatment of human mammary epithelial cells cultures in either well culture or a microfluidic device. Using a single-cell ERK kinase translocation reporter, we observed extended ERK activation in well culture and only transient activity in microfluidic culture. The activity in microfluidic culture resembled that of the control condition, suggesting that shear stress led to the early activity and a loss of autocrine factors dampened extended signaling. Through experimental analysis we identified growth differentiation factor-15 as a candidate factor that led to extended ERK activation through a protein kinase C-α/β dependent pathway. Our results demonstrate that context impacts ERK dynamics and that comparison of distinct contexts can be used to elucidate new aspects of the cell signaling network.

Hyperstable EGF-like bleogen derived from cactus accelerates corneal healing in rats.

Corneal scarring reduces corneal transparency, compromises vision, and is a major cause of vision loss worldwide. Epidermal growth factor (EGF), which is the prototypic member of the EGF receptor (EGFR) agonists, is present in tears to provide repair and regeneration. Recently, we discovered bleogen pB1 in the cactus plant Pereskia bleo and showed that it is a non-canonical and hyperstable EGFR agonist with EGF-like wound healing properties for diabetic rats. Here, we apply bleogen pB1 to accelerate corneal wound healing in rats. To assess the corneal healing effects of bleogen pB1, we induced an acute alkali burn to the right eye of male Wistar rats. After five consecutive ophthalmic applications, fluorescein staining and opacity scores of the bleogen pB1-treated, and the positive control EGF-treated groups improved significantly compared to the saline control group. Immunohistochemical analyses revealed that infiltrated CD68+ macrophages and the expression of the myofibroblast marker alpha smooth muscle actin (α-SMA) were significantly decreased in the bleogen pB1- and the EGF-treated groups. By employing a differential gene expression analysis of bleogen pB1- and EGF-treated keratinocytes through RNA-seq, we demonstrated that bleogen pB1 or EGF treatments can affect the expression of genes associated with inflammatory responses and extracellular matrix remodeling. Taken together, our results indicate that the plant-derived EGFR agonist bleogen pB1 can produce similar effects to those of EGF in accelerating corneal wound healing as well as in reducing persistent inflammation and myofibroblast accumulation in the cornea.