TGF-alpha Research Articles

Efficacy of Supra-HFR in Removing FGF23 and Cytokines: A Single Session Analysis.

Supra hemodiafiltration with reinfusion of the endogenous ultrafiltrate (Supra-HFR) is a dialysis technique used to improve uremic toxin removal in the range of the middle molecular weight molecules. Supra-HFR does not require the preparation and online infusion of high-purity dialysis water because it allows the production of an endogenous ultrafiltrate that undergoes detoxification through an adsorbing resin. We investigated the ability of Supra-HFR to remove fibroblast growth factor 23 (FGF23), interleukin 6 (IL-6), tumor necrosis factor alpha (TNF-alpha), interleukin 8 (IL-8), and transforming growth factor alpha (TGF-alpha) after a single session dialysis in nine patients affected by end stage renal disease (ESRD). The same patients underwent a single session of online hemodiafiltration (OL-HDF) to evaluate possible differences in FGF23 and IL-6 levels. A significant reduction in FGF23 was observed with both Supra-HFR (p=0.001) and OL-HDF. As for TNF-alpha and TGF-alpha, which were measured using Supra-HFR only, their percentage values were significantly lower at the end of dialysis than at the start (p=0.0028 and p=0.03, respectively). This did not change with post-dialysis rebound. Supra-HFR was found to have no effect on IL-6 and IL-8. Interestingly, the removal rate for FGF23 and IL-6 was similar to that observed with OL-HDF. Supra-HFR was not superior to OL-HDF, with suboptimal convective volume in the removal of the molecules tested, especially FGF23, which is considered a large middle molecular weight uremic toxin.

Safety and Efficacy of Enalapril Combined with Metoprolol in Patients of Hypertension Complicated with Coronary Heart Disease: A Randomised Clinical Trial

Introduction: Enalapril, which inhibits Angiotensin-Converting Enzyme (ACE), is a commonly used drug for the treatment of hypertension. Metoprolol, a β-blocker, is widely prescribed for patients with cardiac diseases. Essential hypertension has been identified as an independent risk factor of Coronary Heart Disease (CHD). Until now, there are few studies and reports on the application of the two drugs in patients with Hypertension and Coronary Heart Disease (HCHD). Aim: To assess the safety and efficacy of enalapril combined with metoprolol in patients with HCHD. Materials and Methods: The present single-centre randomised clinical trial was conducted on 109 patients with HCHD at Jingdezhen Municipal First People’s Hospital from April 2018 to January 2020. The participants were randomly divided into into two groups, one group on enalapril (n=58), and the other on enalapril and metoprolol combination (n=51). Patients in both groups were treated for eight weeks. The blood pressure, cardiac function index, serum C-Reactive Protein (CRP), Homocysteine (Hcy), and Matrix Metalloproteinase-9 (MMP-9) levels were compared between the two groups, and the adverse drug reactions of the two groups were statistically analysed using independent sample t-test or Mann-Whitney U test. Results: The mean age of enalapril group was 64.78±7.29 years, and the gender ratio was 1:1.42. The mean age of enalapril/ metoprolol combination group was 63.90±6.91 years, and the gender ratio was 1:0.96. After treatment, both systolic and diastolic blood pressure of the enalapril/metoprolol combination group were significantly lower than that of enalapril group (p<0.05). Enalapril combined with metoprolol significantly improved cardiac function and significantly decreased the levels of serum inflammatory markers (CRP 18.57±3.61 vs 27.50±3.60, p<0.001; Hcy 12.14±2.07 vs 13.83±2.17 mmol L-1, p<0.001; MMP-9 372.35±12.34 vs 436.69±13.89 pg L-1, p<0.001; Transforming Growth Factor ALPHA (TGF-a) 8.18±1.38 vs 10.40±1.44 mmol L-1, p<0.001). The overall rate of adverse reactions was not significantly different between the groups (enalapril/metoprolol combination group vs enalapril group, 15.69% vs 10.34%, p=0.406). Conclusion: The combined treatment of enalapril and metoprolol was more effective than enalapril alone in reducing the inflammatory response, reducing blood pressure, and improving cardiac function in patients with HCHD. This combinational therapeutic strategy may be a better choice for patients with HCHD.

Association of MTHFR, BMP4, TGFA and IRF6 Polymorphisms with Non-Syndromic Cleft lip and Palate in North Indian Patients

Background:Non-Syndromic Cleft Lip and Palate (NSCL/P) is a multifactorial birth defect. The world-wide prevalence of NSCL/P is 1 in 1000 live births; it differs with race, ethnicity and gender. The aim of the present study was to find out the status of candidate gene polymorphisms in NSCL/P cases and its association in phenotype of the patients.Methods:We have screened five polymorphisms in four candidate genes MTHFR (rs1801133, rs1801131) BMP4 (rs17563), TGFA (rs1146297) and IRF6 (rs2235371) by restriction fragment length polymorphism and results were validated by Sanger sequencing. Our dataset consists of 200 NSCL/P cases and 200 healthy controls from the Indian population. Statistical data analysis was performed by SPSS software.Results:MTHFR (rs1801133), BMP4 (rs175563) and TGFA (rs11466297) gene polymorphisms showed significant association with NSCL/P and act as a risk factor in the Indian population (p=<0.05). However, MTHFR (rs1801131), and IRF6 (rs2235371) gene polymorphisms did not show significant association with NSCL/P in the Indian population.Conclusion:The result of the study suggests an association between MTHFR (rs1801133), BMP4 (rs175563) and TGFA (rs11466297) polymorphisms with NSCL/P in Indian population.

Collateral-resistance to estrogen and HER-activated growth is associated with modified AKT, ERα, and cell-cycle signaling in a breast cancer model.

Aim:A model of progressively endocrine-resistant breast cancer was investigated to identify changes that can occur in signaling pathways after endocrine manipulation.Methods:The MCF7 breast cancer model is sensitive to estrogens and anti-estrogens while variant lines previously derived from wild-type MCF7 are either relatively 17β-estradiol (E2 )-insensitive (LCC1) or fully resistant to estrogen and anti-estrogens (LCC9).Results:In LCC1 and LCC9 cell lines, loss of estrogen sensitivity was accompanied by loss of growth response to transforming growth factor alpha (TGFα), heregulin-beta and pertuzumab. LCC1 and LCC9 cells had enhanced AKT phosphorylation relative to MCF7 which was reflected in downstream activation of phospho-mechanistic target of rapamycin (mTOR), phospho-S6, and phospho-estrogen receptor alpha Ser167 [ERα(Ser167)]. Both AKT2 and AKT3 were phosphorylated in the resistant cell lines, but small interfering RNA (siRNA) knockdown suggested that all three AKT isoforms contributed to growth response. ERα(Ser118) phosphorylation was increased by E2 and TGFα in MCF7, by E2 only in LCC1, but by neither in LCC9 cells. Multiple alterations in E2-mediated cell cycle control were identified in the endocrine-resistant cell lines including increased expression of MYC, cyclin A1, cyclin D1, cyclin-dependent kinase 1 (CDK1), CDK2, and hyperphosphorylated retinoblastoma protein (ppRb), whereas p21 and p27 were reduced. Estrogen modulated expression of these regulators in MCF7 and LCC1 cells but not in LCC9 cells. Seliciclib inhibited CDK2 activation in MCF7 cells but not in resistant variants; in all lines, it reduced ppRb, increased p53 associated responses including p21, p53 up-regulated modulator of apoptosis (PUMA), and p53 apoptosis-inducing protein 1 (p53AIP1), inhibited growth, and produced G2/M block and apoptosis.Conclusions:Multiple changes occur with progression of endocrine resistance in this model with AKT activation contributing to E2 insensitivity and loss of ERα(Ser118) phosphorylation being associated with full resistance. Cell cycle regulation is modified in endocrine-resistant breast cancer cells, and seliciclib is effective in both endocrine-sensitive and resistant diseases.

Genome-wide Studies of knee Osteoarthritis: Review

Background. Knee osteoarthritis (OA) is a multifactorial disease resulting from the interaction of many environmental, epigenetic and genetic risk factors, and the latter account for 40% to 65%. Genetic bases of the knee OA based on genome-wide association study (GWAS) are being actively studied by many scientific teams around the world. At the same time, the results obtained are often contradictory and ambiguous, as for the conducted replicative studies of knee OA. This dictates the need for additional replicative studies in various populations, including populations of Russia. The aim of the study was to analyze genome-wide studies of knee OA and to establish GWAS-significant polymorphic loci associated with OA. Materials and Methods. The search for publications was carried out in the electronic databases PubMed, PubMedCentral, eLIBRARY, in the GWAS catalog for the period from 2008 to the present by the keywords: knee osteoarthritis, GWAS studies, candidate genes. Results. Firstly, for the period from 2008 to 2021, 15 genome-wide studies of knee OA were performed (8 GWAS, 6 meta-analyzes of GWAS data, 1 study a combination of GWAS and meta-analysis of GWAS data), as a result of which 78 polymorphic loci were found associated with the risk of developing osteoarthritis of the knee joint at p510-08. Second ly, the vast majority of these loci (62 out of 78 SNPs, 79%) showed GWAS-significant associations with OA in meta-analyzes of GWAS data and only 16 loci (21%) in GWAS studies. Third ly, almost 95% of GWAS-significant loci for knee OA (74 SNPs) were found in samples of Caucasian origin. Fourthly, 21 out of 78 GWAS-significant SNPs are associated with isolated knee OA, and 57 SNPs are associated with knee, hip and hand OA (mixed sample). Fifth, all genome-wide studies of knee OA and meta-analyzes of GWAS data were carried out abroad on samples from various foreign populations, and samples from the Russian Federation were not included in these studies. Sixth, only two GWAS-significant polymorphic loci for OA (rs143384 of the GDF5 gene for knee OA isolated localization and rs3771501 of the TGFA gene for OA of any localization) were replicated at the whole genome level of significance (p510-08) in two different studies. Conclusion. The main genome-wide studies of knee OA were reviewed and GWAS-significant polymorphisms associated with OA were identified. The obtained materials on GWAS-significant loci can be used both in the selection of polymorphisms in replicative studies of OA in various populations of Russia, and for expanding the understanding of the molecular genetic mechanisms of the disease development.

Role of Epidermal Growth Factor (EGFR) in Oral Cancer

Oral cancer is a pronounced disease and has the sixth-highest incidence among all the cancers occurring worldwide. Epidermal Growth Factor (EGFR) is of most prognostic significance among various markers found in oral squamous cell carcinoma. But no established criterion is universally used to evaluate EGFR expression. EGFR is known to perform a significant role in oral cancer development, and its overexpression dictates the poor clinical course of cancer. It is considered to be not only a valuable prognostic biomarker but also a promising therapeutic target in cancer treatment. Stimulation of EGFR enhances cancer processes, including increased cell division, neovascularization, invasion/metastasis, and escape from apoptosis. Increased EGFR levels are also observed in other cancers of the body. EGFR detection in OSCC can fulfill multiple roles in cancer diagnostics, such as early-stage, prognosis assessment, and treatment plan. The EGFR is a proto-oncogene activated at the cell membrane surface by transforming growth factor-α serves to advance cellular proliferation in cancer tissue. The literature demonstrates that EGFR is an important target for anti-cancer drugs in an advanced stage of head and neck cancers. Hence, understanding EGFR is essential to learning the development cascade of cancer.

Transforming Growth Factor-α Acts in Hypothalamic Paraventricular Nucleus to Upregulate ERK1/2 Signaling and Expression of Sympathoexcitatory Mediators in Heart Failure Rats

Activation of epidermal growth factor receptor (EGFR) tyrosine kinase is associated with increased extracellular signal-regulated kinase (ERK) 1/2 signaling in the hypothalamic paraventricular nucleus (PVN), which contributes to the sympathetic excitation in heart failure (HF). Transforming growth factor (TGF)-α is a major endogenous ligand for EGFR. The present study sought to determine whether TGF-α increases in the PVN in HF and promotes the activation of EGFR to increase ERK1/2 activity. Male rats received bilateral PVN microinjections of an EGFR siRNA or a scrambled siRNA followed by an intracerebroventricular (ICV) injection of TGF-α or vehicle one week later. In rats pretreated with the scrambled siRNA, ICV TGF-α increased phosphorylated (p-) EGFR and upregulated the expression of p-ERK1/2 and mRNA levels of proinflammatory cytokines (PICs) and renin-angiotensin system (RAS) components in the PVN, when compared with the untreated age-matched control rats. These responses to ICV TGF-α were significantly attenuated in rats pretreated with the EGFR siRNA. Furthermore, bilateral PVN microinjection of a TGF-α siRNA in HF rats significantly decreased the elevated levels of TGF-α, p-EGFR, p-ERK1/2 and the mRNA expression of PICs and RAS components in the PVN, compared with the HF rats treated with a scrambled siRNA. The TGF-α siRNA-treated HF rats also exhibited lower plasma norepinephrine levels and improved peripheral manifestations of HF. These data suggest that TGF-α expression is upregulated in the PVN in HF and induces the activation of EGFR-mediated ERK1/2 signaling to augment the inflammation and RAS activity that drives sympathetic excitation in HF.

Relationship between reduced IL-10 expression and improvement of periodontal conditions of periodontal and diabetic patients

OBJECTIVE: Th is study aims to determine the profi le of infl ammatory proteins in unstimulated whole saliva of periodontal patients with or without type 2 diabetes two months aft er non-surgical periodontal therapy. METHODS: Fift y-six patients were enrolled: PD group with 21 patients, PD + DM group with 20 patients and control group with 15 patients. Clinical examination and glycated hemoglobin (HbA1c) measurement were performed before and two months aft er periodontal therapy. Saliva samples were collected to assess 39 salivary proteins using Multiplex Cytokine Profi ling Assay. RESULTS: Th ere were signifi cant reduction two months aft er periodontal therapy for salivary levels of interleukin (IL)-1β, IL-17, IL-10, eotaxin, and monocyte chemoattractant protein (MCP)-3 in PD group, and for salivary levels of IL-1ra, IL-10 and transforming growth factor-alpha (TGF-α) in PD+DM group. Clinical parameters improved signifi – cantly aft er periodontal therapy in both diseased groups, and HbA1c levels in PD+DM group (p&lt;0.05). CONCLUSIONS: Non-surgical periodontal therapy was eff ective in producing shortterm clinical benefi ts to the patients with a signifi cant reduction of important infl ammatory markers related to the pathogenesis of the disease. Among the studied cytokines only IL-10 shared similar outcome in the diseased groups, suggesting its potential applicability as a salivary biomarker for monitoring periodontitis associated or not to diabetes and infl ammation during the early response to therapy.

A molecular mechanism for the generation of ligand-dependent differential outputs by the epidermal growth factor receptor

The epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase that couples the binding of extracellular ligands, such as EGF and transforming growth factor-α (TGF-α), to the initiation of intracellular signaling pathways. EGFR binds to EGF and TGF-α with similar affinity, but generates different signals from these ligands. To address the mechanistic basis of this phenomenon, we have carried out cryo-EM analyses of human EGFR bound to EGF and TGF-α. We show that the extracellular module adopts an ensemble of dimeric conformations when bound to either EGF or TGF-α. The two extreme states of this ensemble represent distinct ligand-bound quaternary structures in which the membrane-proximal tips of the extracellular module are either juxtaposed or separated. EGF and TGF-α differ in their ability to maintain the conformation with the membrane-proximal tips of the extracellular module separated, and this conformation is stabilized preferentially by an oncogenic EGFR mutation. Close proximity of the transmembrane helices at the junction with the extracellular module has been associated previously with increased EGFR activity. Our results show how EGFR can couple the binding of different ligands to differential modulation of this proximity, thereby suggesting a molecular mechanism for the generation of ligand-sensitive differential outputs in this receptor family.

Regulation of osteoarthritis development by ADAM17/Tace in articular cartilage.

A disintegrin and metalloproteinase 17 (Adam17), also known as TNFα-converting enzyme (Tace), is a membrane-anchored protein involved in shedding of TNF, IL-6 receptor, ligands of epidermal growth factor receptor (EGFR), and Notch receptor. This study aimed to examine the role of Adam17 in adult articular cartilage and osteoarthritis (OA) pathophysiology. Adam17 expression was examined in mouse knee joints during OA development. We analyzed OA development in tamoxifen-inducible chondrocyte-specific Adam17 knockout mice of a resection of the medial meniscus and medial collateral ligament (medial) model, destabilization of the medial meniscus (DMM) model, and aging model. We analyzed downstream pathways by in vitro experiments, and further performed intra-articular administration of an Adam17 inhibitor TAPI-0 for surgically induced mouse OA. Adam17 expression in mouse articular cartilage was increased by OA progression. In all models, Adam17 knockout mice showed ameliorated progression of articular cartilage degradation. Adam17 knockout decreased matrix metallopeptidase 13 (Mmp13) expression in both in vivo and in vitro experiments, whereas Adam17 activation by phorbol-12-myristate-13-acetate (PMA) increased Mmp13 and decreased aggrecan in mouse primary chondrocytes. Adam17 activation enhanced release of soluble TNF and transforming growth factor alpha, a representative EGF ligand, from mouse primary chondrocytes, while it did not change release of soluble IL-6 receptor or nuclear translocation of Notch1 intercellular domain. Intra-articular administration of the Adam17 inhibitor ameliorated OA progression. This study demonstrates regulation of OA development by Adam17, involvement of EGFR and TNF pathways, and the possibility of Adam17 as a therapeutic target for OA.

Nodal/SMAD2 and Bone Morphogenic Protein 4 RNA Expression in Microgravity. Implications for Spacial Orientation Signaling in Hematopoesis and the Implications for the Hematosuppressive Effects of Space Flight

Introduction: Nodal proteins are members of the TGF alpha family which direct right left orientation in the developing embryo. Nodal/SMAD2 signaling controls BMP4 expression and with canonical Wnt signaling, regulates the posteriorization of the primitive streak, development of the mesoderm, development of early hematopoetic stem cells, and the property of self renewal in adult hematopoiesis. This study investigated the extent that microgravity alters expression of these spatially orienting proteins of BMP4, Nodal, Wnt, and SMAD2 thus influencing the anemia, thrombocytopenia, and immune alteration associated with space flight.Method: The Gene Expression Omnibus Database (GEO) of the National Institutes of Health was accessed to define RNA expression of Nodal, SMAD2, Wnt and BMP4 in hematopoetic cells in microgravity. Initial queries for “Low Gravity” studies resulted in 565 investigations within the GEO Omnibus. Of the 565 studies, the GSE136939, GSE101102, and the GSE101309 datasets were explored which assessed the lymphoid and macrophage cell lines TK6, U937, and Jurkat. RNA expression with normal and microgravity environments were compared. In the U937 cells hypergravity environments were also studied. Nodal, Lefty, Wnt, BMP4, as well as the NODAL signaling cascade proteins of SMAD 1/5, SMAD2, SMAD3, and the inhibitory SMAD 7. Affymetrix Expression Arrays were utilized for the TK^ and Jurkat cells assays. The U937 cell lines were assayed with NimbleGen Human Gene Assay Arrays.Results: Significant decreases in BMP4 expression were seen in microgravity environments (P value, <0.005) where SMAD2 expression was significantly elevated (P value <0.05) in microgravity. Conversely, hypergravity effects on U937 cells resulted in a reverse expression pattern. (BMP4 expression increased while SMAD2 decreased (P value <0.01) ). Comparisons of 14, 12, and 4 expression assays were made in U937, Jurkat, and TK6 cells respectively. LEFTY, NODAL, Wnt, SMA1/5/3/7 expression assays all showed changes in expression, but the small sample numbers resulted in poor statistical power and insignificant P values to detect differences in the RNA expression assessed. Comparisons of the gravitational pressures and time spent in these environments for the cellular cultures resulted in insignificant statistical differences.Conclusions: Microgravity induces decreases in BMP4 RNA expression and increases in SMAD2 RNA expression in TK6, U937, and Jurkat cell as defined by the GSE101309, GSE101102 and GSE136939 datasets. BMP4 is required for stem cell posteriorization, HSC number, and adult hematopoetic self renewal. This diminishment in BMP4 expression in microgravity may result in diminished HSC number and self renewal capacity. This qualitative change in gene expression in microgravity and may be a significant component of the anemia, thrombocytopenia, and immune alterations of space flight. Moreover such changes in RNA expression in microgravity implicates the importance of hematopoetic spacial orientation signals in normal hematopoiesis. Therapeutic targeting of the BMP/SMAD2 pathway in refractory multilineage hematopoetic diseases may prove to be fruitful. Statistical interpretation of the data is hindered by the small number of samples, the cell type, hematopoetic maturation stage of the samples, and microenvironmental differences in the studies utilized in these datasets. If this result is confirmed in future and larger studies, it would have significant biologic and therapeutic implications for long term space flight and the developing concept of spacial hematopoiesis. DisclosuresNo relevant conflicts of interest to declare.

Redundant roles of EGFR ligands 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.

Magnitude of Ubiquitination Determines the Fate of Epidermal Growth Factor Receptor Upon Ligand Stimulation

Receptor tyrosine kinases (RTK) bind growth factors and are critical for cell proliferation and differentiation. Their dysregulation leads to a loss of growth control, often resulting in cancer. Epidermal growth factor receptor (EGFR) is the prototypic RTK and can bind several ligands exhibiting distinct mitogenic potentials. Whereas the phosphorylation on individual EGFR sites and their roles for downstream signaling have been extensively studied, less is known about ligand-specific ubiquitination events on EGFR, which are crucial for signal attenuation and termination. We used a proteomics-based workflow for absolute quantitation combined with mathematical modeling to unveil potentially decisive ubiquitination events on EGFR from the first 30 seconds to 15 minutes of stimulation. Four ligands were used for stimulation: epidermal growth factor (EGF), heparin-binding-EGF like growth factor, transforming growth factor-α and epiregulin. Whereas only little differences in the order of individual ubiquitination sites were observed, the overall amount of modified receptor differed depending on the used ligand, indicating that absolute magnitude of EGFR ubiquitination, and not distinctly regulated ubiquitination sites, is a major determinant for signal attenuation and the subsequent cellular outcomes.

Pilot Study Analysis of Serum Cytokines to Differentiate Pediatric Septic Arthritis and Transient Synovitis.

In pediatric patients, the presentation of the nontraumatic acutely painful joint/limb poses a diagnostic dilemma due to the similarity of presentations of the most likely diagnoses [septic arthritis (SA), transient synovitis (TS), osteomyelitis]. Current tools employed to differentiate these diagnoses rely on nonspecific inflammatory markers, radiologic imaging, and arthrocentesis. Diagnostic algorithms utilizing these clinical, radiographic, and biochemical parameters have produced conflicting results. The purpose of this study was to identify a serum-based inflammatory signature which can differentiate SA from TS in pediatric patients. Serum samples were collected from 22 pediatric patients presenting with joint/extremity pain whose working diagnosis included SA or TS. Each sample was analyzed for serum abundance of 72 distinct biomarkers and cytokines using enzyme linked immunosorbent assay based arrays. Linear discriminant analysis was performed to identify a combinatorial biomarker panel to predict a diagnosis of SA or TS. Efficacy of the biomarker panel was compared with definitive diagnoses as based on laboratory tests, arthrocentesis results, and clinical scenario. At the time of presentation: (1) mean erythrocyte sedimentation rate in the SA group was 56.6 mm/h and 12.4 mm/h in the TS group (P<0.001), (2) mean C-reactive protein was 55.9 mg/dL in the SA group and 13.7 mg/dL in the TS group (P=0.12), and (3) mean white blood cell was 10.9 k/mm3 in the SA group and 11.0 k/mm3 in the TS group (P=0.95). A combined panel of 72 biomarkers was examined using discriminant analysis to identify a limited set of predictors which could accurately predict whether a patient was diagnosed with SA or TS. A diagnostic algorithm consisting of transforming growth factor alpha, interleukin (IL)-7, IL-33, and IL-28A serum concentration correctly classified 20 of the 22 cases with a sensitivity and specificity of 90.9% (95% confidence interval: 73.9%-100.0%). This study identifies a novel serum-based 4-cytokine panel that accurately differentiates SA from TS in pediatric patients with joint/limb pain. Level II-diagnostic study.

Molecular-genetic mechanisms of the signal cascade RAS-RAF-MEK-ERK associated with the development of the tumor process and the purpose of targeted drugs for colorectal cancer

Colorectal cancer occupies one of the leading positions in the world in the structure of cancer incidence. The vital processes of cancer cells largely depend on the production of growth factors and their receptors. One of these is epidermal growth factor (EGFR), which is a tyrosine kinase receptor for cell membranes. Normally, binding of EGFR ligands and transforming growth factor alpha (TGFα) induces receptor activation, which triggers ERK and PI3K signaling pathways that control cell proliferation, migration, invasion, and many other processes. It was found that in 80% of cases, colorectal cancer occurs as a result of EGFR overexpression, which leads to increased growth and division of tumor cells due to hyperactivation of the RAS-RAF-MEKERK signaling cascade. The RAS-RAF-MEK-ERK cascade is a pathway that regulates cell proliferation, cell cycle, and cell migration. In the development of human cancer, mutations of the RAS/RAF family are most often the cause of dysregulation of signal transduction through this pathway. According to current data, about a third of all malignant neoplasms are associated with mutations in the genes of the RAS family, which include HRAS, KRAS, NRAS, RRAS, and other homologous proteins. Proteins of the RAS family are involved in the activation of tyrosine kinase signaling pathways, which leads to gene mutations. This process determines proliferative activity, ability to differentiate, metastasis, avoidance of apoptosis, induction of angiogenesis. Permanent RAS activation leads to malignant cell degeneration. Thus, the expression and mutation of the EGFR gene are associated with various variants of tumor progression and poor prognosis in malignant neoplasms of various localizations. Over the past decades, significant advances have been made in the treatment of metastatic colorectal cancer. However, the expansion of the spectrum of effective anticancer drugs also creates a number of difficulties in choosing the optimal drug therapy regimens in patients with metastases of colorectal cancer.

The Aryl Hydrocarbon Receptor-Dependent TGF-α/VEGF-B Ratio Correlates With Disease Subtype and Prognosis in Multiple Sclerosis.

ObjectiveTo evaluate the aryl hydrocarbon receptor (AHR)-dependent transforming growth factor alpha (TGF-α)/vascular endothelial growth factor B (VEGF-B) ratio, which regulates the effects of metabolic, dietary, and microbial factors on acute and chronic CNS inflammation, as a potential marker in multiple sclerosis (MS).MethodsTGF-α, VEGF-B, and AHR agonistic activity were determined in serum of 252 patients with relapsing-remitting (RR) MS, primary and secondary progressive MS, as well as during active disease (clinically isolated syndrome [CIS] and RRMS relapse).ResultsThe TGF-α/VEGF-B ratio and AHR agonistic activity were decreased in all MS subgroups with a stable disease course as compared to controls. During active CNS inflammation in CIS and RRMS relapse, the TGF-α/VEGF-B ratio and AHR agonistic activity were increased. Conversely, in patients with minimal clinical impairment despite long-standing disease, the TGF-α/VEGF-B ratio and AHR agonistic activity were unaltered. Finally, the TGF-α/VEGF-B ratio and AHR agonistic activity correlated with neurologic impairment and time to conversion from CIS to MS.ConclusionsThe AHR-dependent TGF-α/VEGF-B ratio is altered in a subtype, severity, and disease activity–specific manner and correlates with time to conversion from CIS to MS. It may thus represent a novel marker and serve as additive guideline for immunomodulatory strategies in MS.Classification of EvidenceThis study provides Class III evidence that serum levels of AHR, TGF-α, and VEGF-B distinguish subtypes of MS and predict the severity and disease activity of MS.

Triple therapy with osimertinib, bevacizumab and cetuximab in EGFR-mutant lung cancer with HIF-1α/TGF-α expression.

Osimertinib, a third generation epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, is the standard treatment for patients with lung cancer harboring EGFR T790M; however, acquired resistance is inevitable due to genetic and epigenetic changes in cancer cells. In addition, a recent randomized clinical trial revealed that the combination of osimertinib and bevacizumab failed to exhibit superior progression-free survival compared with osimertinib alone. The present study aimed to investigate the effect of triple therapy with osimertinib, bevacizumab and cetuximab in xenograft tumors with different initial tumor volumes (conventional model, 200 mm3 and large model, 500 mm3). The results demonstrated that osimertinib significantly inhibited tumor growth in both the conventional and large models; however, maximum tumor regression was attenuated in the large model in which hypoxia-inducible factor-1α (HIF-1α) and transforming growth factor-α (TGF-α) expression levels increased. Although the combination of osimertinib and bevacizumab exerted a greater inhibitory effect on tumor growth compared with osimertinib in the conventional model, the effect of this combination therapy was attenuated in the large model. TGF-α attenuated sensitivity to osimertinib in vitro; however, this negative effect was counteracted by the combination of osimertinib and cetuximab, but not osimertinib and bevacizumab. In the large xenograft tumor model, the triple therapy induced the greatest inhibitory effect on tumor growth compared with osimertinib alone and its combination with bevacizumab. Clinical trials of the triple therapy are required for patients with lung cancer with EGFR mutations and HIF-1α/TGF-α.

Abstract 1817: Epidermal growth factor receptor alters major histocompatibility complex class I expression on pancreatic cancer cells

Abstract Pancreatic cancer is one of the deadliest neoplasms, with a poor 5-year survival rate of just 10%. The ability of pancreatic cancer cells to evade the immune system hinders an anti-tumor response and contributes to the poor survival rate. In some cases, down regulation of major histocompatibility complex (MHC) class I cell-surface expression aids in immune evasion by preventing endogenous tumor antigens from being presented to cytotoxic T cells. The goal of this study is to evaluate the role epidermal growth factor receptor (EGFR) plays in the regulation of MHC class I expression by pancreatic cancer cells. Pancreatic cancer cell lines were treated with epidermal growth factor (EGF) or transforming growth factor alpha (TGF-alpha) to activate EGFR or with the small molecule drug erlotinib to inhibit EGFR. Immunoprecipitation, western blot analysis, and flow cytometry were used to analyze cells' MHC class I expression post treatment. Human pancreatic cancer cell lines (BxPC-3, T3M4, S2-013, Panc-1) were found to decrease MHC class I expression after EGFR activation by EGF. However, activation of EGFR via TGF-alpha led to differential effects on MHC class I expression in a cell line-dependent manner. EGFR inhibition by erlotinib led to increased MHC class I expression by greater than 2.5-fold on both human and mouse pancreatic cancer cells. Mechanistic studies were done to evaluate the role of src homology 2 phosphatase (SHP2) in EGFR's regulation of MHC class I in pancreatic cancer cells, since in head and neck cancer cells SHP2 can be phosphorylated by activated EGFR and subsequently dephosphorylate STAT1 and reduce MHC class I expression. Following SHP2 knockdown by siRNA transfection, a slight increase in MHC class I expression was observed on pancreatic cancer cells, suggesting SHP2 has a minor down-regulatory role in MHC class I expression in these cells. EGFR was found to co-immunoprecipitate with MHC class I, indicating that EGFR might directly affect MHC class I expression by facilitating its endocytosis. However, after EGFR inhibition, no change in the MHC class I cell surface turnover rate was observed. Other potential mechanisms for EGFR regulation of MHC class I expression are currently being evaluated. Our discoveries defining a role for activated EGFR in reducing MHC class I expression and revealing that EGFR inhibitors can boost MHC class I levels on cancer cells could facilitate the adoption of new, more effective therapeutic tactics that combine the use of EGFR inhibitors with immunotherapies to augment patients' immune systems for effective targeting and killing of cancer cells. Citation Format: Shelby M. Knoche, Joyce C. Solheim. Epidermal growth factor receptor alters major histocompatibility complex class I expression on pancreatic cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1817.

Phorbol Diesters and 12-Deoxy-16-hydroxyphorbol 13,16-Diesters Induce TGFα Release and Adult Mouse Neurogenesis.

A small library of phorbol 12,13-diesters bearing low lipophilicity ester chains was prepared as potential neurogenic agents in the adult brain. They were also used in a targeted UHPLC-HRMS screening of the latex of Euphorbia resinifera. Two new 12-deoxy-16-hydroxyphorbol 13,16-diesters were isolated, and their structures were deduced using two-dimensional NMR spectroscopy and NOE experiments. The ability of natural and synthetic compounds to stimulate transforming growth factor alpha (TFGα) release, to increase neural progenitor cell proliferation, and to stimulate neurogenesis was evaluated. All compounds that facilitated TGFα release promoted neural progenitor cell proliferation. The presence of two acyloxy moieties on the tigliane skeleton led to higher levels of activity, which decreased when a free hydroxyl group was at C-12. Remarkably, the compound bearing isobutyryloxy groups was the most potent on the TGFα assay and at inducing neural progenitor cell proliferation in vitro, also leading to enhanced neurogenesis in vivo when administered intranasally to mice.

TGFβ1, MMPs and cytokines profiles in ocular surface: Possible tear biomarkers for pseudoexfoliation.

Pseudoexfoliation (PXF) is a unique form of glaucoma characterized by accumulation of exfoliative material in the eyes. Changes in tear profile in disease stages may give us insights into molecular mechanisms involved in causing glaucoma in the eye. All patients were categorized into three main categories; pseudoexfoliation (PXF), pseudoexfoliation glaucoma (PXG) and cataract, which served as control. Cytokines, transforming growth factor β1 (TGFβ1), matrix metalloproteases (MMPs) and fibronectin (FN1) were assessed with multiplex bead assay, enzyme-linked immunosorbent assay (ELISA), gelatin zymography, and immunohistochemistry (IHC) respectively in different ocular tissues such as tears, tenon's capsule, aqueous humor (AH) and serum samples of patients with PXF stages. We found that TGFβ1, MMP-9 and FN1 protein expression were upregulated in tears, tenon's capsule and AH samples in PXG compared to PXF, though the MMP-9 protein activity was downregulated in PXG compared with control or PXF. We have also found that in PXG tears sample the fold change of TGF-α (Transforming Growth Factor-α), MDC (Macrophage Derived Chemokine), IL-8 (Interleukin-8), VEGF (Vascular Endothelial Growth Factor) were significantly downregulated and the levels of GM-CSF (Granulocyte Macrophage Colony Stimulating Factor), IP-10 (Interferon- γ produced protein-10) were significant upregulated. While in AH; IL-6 (Interleukin-6), IL-8, VEGF, IFN-a2 (Interferon- α2), GRO (Growth regulated alpha protein) levels were found lower and IL1a (Interleukin-1α) level was higher in PXG compared to PXF. And in serum; IFN-a2, Eotaxin, GM-CSF, Fractalkine, IL-10 (Interleukin-10), IL1Ra (Interleukin-1 receptor antagonist), IL-7 (Interleukin-7), IL-8, MIP1β (Macrophage Inflammatory Protein-1β), MCP-1 (Monocyte Chemoattractant Protein-1) levels were significantly upregulated and PDGF-AA (Platelet Derived Growth Factor-AA) level was downregulated in the patients with PXG compared to PXF. Altered expression of these molecules in tears may therefore be used as a signal for onset of glaucoma or for identifying eyes at risk of developing glaucoma in PXF.