EGF Levels Research Articles

PCN68 COST EFFECTIVENESS OF CIMAVAX EGF VS. STANDARD-OF-CARE AS MAINTENANCE TREATMENT FOR ADVANCED NSCLC THAT EXPRESSES HIGH LEVELS OF EGF IN COLOMBIA

The aim of the study was to evaluate the cost effectiveness of CIMAvax-EGF, compared with standard-of-care (SoC) as maintenance treatment in patients with non-small-cell lung cancer (NSCLC) that expresses high levels of EGF, from a Colombia third-party public healthcare payer perspective. We conducted a markov model with a cycle length of three months and a time horizon of four years. Parametric survival models were fitted to Kaplan–Meier estimates of overall survival from a randomized clinical trial (patients aged ≥18 years with stage IIIB – IV NSCLC, ECOG 0-2, who received systemic chemotherapy). Progression free survival were validated from registry data. Costs (COP, year 2016 values) for drug acquisition/administration and clinical management were included. Costs and outcomes were discounted at 5% per year. For the base-case scenario data was evaluated for patients with squamous NSCLC. An alternative scenario was evaluated for all patients with NSCLC comparing CIMAvax-EGF to other maintenance therapies. A deterministic and probabilistic sensitivity analyses were performed. In the base-case scenario, CIMAvax resulted in an expected gain of survival. The incremental cost per LY gain was COP 30.132.000/LY. The cost-effectiveness threshold in Colombia established for the year 2016 is approximately COP 43,500,000 / LY. So, it is considered that CIMAvax-EGF is a cost-effective therapy. The sensitivity analysis showed that the cost of second-line therapy is the variable that most affects the mode. In 95% of the cases evaluated, CIMAvax-EGF was the most cost-effective therapy. CIMAvax-EGF is a cost-effective option compared with SoC as maintenance treatment in adults with NSCLC expressing high levels of EGF.

Association of parameters of the stress system and the immunity in patients with chronic pyelonephritis in dependence on the duration of the disease and the frequency of relapses

The immune response in chronic pyelonephritis (CP) is a compensatory-adaptive response of the growing organism, which determines the character of the course and prognosis of the disease. The aim of the study was the investigation of relationships of the immune and endocrine systems in dependence on the duration of the course and frequency of recurrence of CP in children. There were observed a total of 168 CP children patients at the age of 6-16 years, in a state of clinical and laboratory remission. The reference group was consisted of 20 apparently healthy children of the same age. During the remission of CP duration of the disease and the frequency of CP relapses were established to influence on parameters of the stress-realizing and stress-limiting systems. There was revealed a direct correlation between the levels both of ACTH and cortisol with the number of CP relapses, which were dependent, in turn, on their frequency and duration of disease. There was revealed the inversion of the rhythm of melatonin secretion, the more pronounced the longer course of CP and their relapses occur more frequently. There was revealed adaptive directedness of the impact of melatonin on the immune system, as evidenced by the revealed negative correlations of the peak value of night 6- sulfatoxymelatonin (6-COMT) with TGF-β1 and IL-17. The increasing of the duration of CP was accompanied by an elevation in the concentrations of TGF-β1 and its positive correlation with ACTH levels, as well as a significant decrease in IL-10, IGF-1 and EGF levels. The authors suggest that in CP patients there is formed the imbalance between stress-realizing and stress-limiting systems, the more pronounced the longer course of CP and its relapses occur more often.

THE PRETERM INFANT GUT MICROBIOME, HUMAN MILK VOLUME AND IMMUNOBIOLOGY, AND GROWTH

We conducted a study of gut microbiome in very low birth weight (VLBW) infants along with measures of milk feedings (type and volume) and milk sIgA, cytokines and chemokines for six weeks after admission in 78 infants. Stools were collected weekly, and human milk collected daily and pooled weekly for a panel of immune markers. Mothers of these infants gave informed consent and the study was approved by the university IRB. Mean levels of sIgA, EGF, MIP1α, IP10, MCP1, IL‐8, IL‐4, IL‐6, IL‐10 and TNFα were measured each week for six weeks by MagPix.4.2. Stool DNA was extracted using MoBio kits, the V3–V4 region of the 16S rRNA gene was amplified and then sequenced on an Illumina MiSeq.These infants had a mean gestation age at birth of 28 weeks, 3 days, and a mean birth weight of 1078 Gms. The mean time to full enteral feeding was 12 days, and they were fed mothers own milk (MOM), donor milk or preterm infant formula in varying amounts. The infant stool demonstrated marked dysbosis over time, with a dominance of Gram negative Gammaproteobacteria. Mean levels of sIgA, EGF, MIP1α, IP10, MCP1, IL‐8, IL‐4, IL‐6, IL‐10 and TNFα were measured each week for six weeks. Data reported here are mean Shannon diversity, and mean number of operational taxonomic units (OTUs) over the first six weeks of admission to the NICU, the ratio of human milk volumes to total feed volumes, the mean concentrations of milk immune markers over the 6 weeks of measurement, and the relationship between growth and the microbiome in these infants.Shannon diversity significantly increased with increasing corrected gestational age (cGA) (Multiple R2: 0.09221, p= 1.28e‐08). Adjusting for cGA during the NICU stay, the alpha diversity also showed weak but significant correlations with overall rate of weight gain (Multiple R2: 0.144, p=0.00208) and total ratio of mother's own milk (Multiple R2: 0.1097, p = 0.024918).Over the 6 weeks of admission, the mean number of fecal OTUs was correlated with mean milk IL‐4 (pg/ml) (r=.25, p=.03), but no other milk immune markers. The time to enteral feeding was inversely correlated to the number of OTUs (r=‐.24, p=.036). Lower diversity was observed in infants with lower gestational age (t=.31, p=.006), sepsis (t=2.43, p=.017), and days on oxygen (r=.23, p=.04).This is the first study to show that growth of VLBWs may be influenced by the microbiome in these early weeks of life. There were few relationships between specific immune markers and gut microbiome, but the amount of MOM an infant receives may partially ameliorate the dysbiosis.Future studies are planned to investigate the growth/nutrition/microbiome relationship in VLBWs as they grow and develop into early school age.Support or Funding InformationNIH R01NR15546This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Association of EGF A61G polymorphism and EGF expression with type 2 diabetes mellitus in Indian population

BackgroundEpidermal Growth Factor (EGF) plays critical role in regeneration of pancreatic β-cells and insulin secretion. The deficiency in EGF expression affects pancreatic β-cell's proliferation, maintenance and its function like insulin secretion which may result in individual susceptibility to Type 2 Diabetes Mellitus (T2DM). EGF A61G is a functional SNP that modulate EGF levels and eventually affects activity of β-cells. ObjectiveThe present study explores EGF A61G polymorphism (rs4444903) and correlates it with the risk of developing T2DM and mRNA expression. MethodsThe study included 90 subjects, T2DM patients (n = 30) and healthy controls (n = 60). Blood sample was collected from subjects to study EGF A61G polymorphism by PCR-RFLP and EGF mRNA expression level by Real-time PCR. ResultsThe EGF gene A61G polymorphism was found to be in significant association with T2DM. A/G genotype (OR = 4.07, 95%CI = 1.303 to 12.71, p = 0.0206) and ‘A’ allele (OR = 1.91, 95%CI = 1.017 to 3.568, p = 0.043) were significantly associated with risk of T2DM. The prevalence of allele ‘A’ was found to be higher in diabetic males and females (52.5% and 55% respectively). As compared to healthy subjects EGF mRNA expression levels showed significant down-regulation in T2DM patients (p = 0.005). The relative EGF mRNA expression in A/A and A/G genotype was significantly lower than G/G genotype. ConclusionThe present study suggests that EGF gene A61G polymorphism may be a genetic risk factor for susceptibility of T2DM in Indian population. However, this marker deserves further investigation using more sample size.

Accuracy of Urinary Epidermal Growth Factor to Creatinine Ratio to Predict 24-Hour Urine Epidermal Growth Factor and Interstitial Kidney Fibrosis in Patients with IgA Nephropathy.

Urinary levels of EGF may be a noninvasive biomarker of the degree of interstitial fibrosis. However, all the available data are based on studies that examined the EGF/creatinine ratio in spot urine samples. The agreement between EGF/creatinine ratio and 24-hours EGF excretion has not been analyzed, neither has it been established which of these two measurements is a better predictor of the degree of interstitial fibrosis. To investigate whether the EGF/creatinine ratio can predict 24-hours EGF, and which of these two measures is a better predictor of interstitial fibrosis in patients with IgA nephropathy (IgAN). This is a cross-sectional study including 80 patients with IgAN. EGF levels were measured by ELISA in spot second-morning and 24-hours urine samples. We analyzed the concordance between these two measures and their respective ability to predict interstitial kidney fibrosis. The intraclass correlation coefficient between 24-hours and spot EGF/creatinine was 0.63 (95% CI: 0.54 - 0.70), bias was 2.7 µg/mL (95% CI: 2.1 - 7.5). Passing-Bablok regression did not show a significant deviation from linearity (p = 0.72). Bland-Altman showed a systematic and proportional error between both EGF measures. Spot EGF/creatinine ratios overestimated the 24-hours EGF at low excretion values and underestimated it at high excretion values. In univariate analyses, 24-hours excretion of EGF was a better predictor of interstitial fibrosis than spot EGF/creatinine ratio (R2: 0.43 vs. 0.30, p = 0.000). In multivariate analyses, the 24-hours excretion of EGF plus GFR, significantly improved the prediction of interstitial fibrosis when compared with GFR alone (R2: 0.52 vs. 0.39, p = 0.000). When spot-urine EGF was introduced instead of the 24-hours excretion, the model was statistically significant but had a lower predictive capacity (R2: 0.46 spot EGF/creatinine vs. R2: 0.52 24-hours EGF excretion, p = 0.000). The 24-hours excretion of EGF should be considered as the first-choice measure to estimate the interstitial fibrosis. The EGF/creatinine ratio cannot accurately estimate the total EGF excretion of but it also improves the estimation of the fibrosis surface, and, consequently, could be an alternative whenever 24-hours urine samples cannot be obtained.

The Prognostic Value of Serum Cytokines in Patients with Acute Ischemic Stroke.

The inflammatory response is an unavoidable process and contributes to the destruction of cerebral tissue during the acute ischemic stroke (AIS) phase and has not been addressed fully to date. Insightful understanding of correlation of inflammatory mediators and stroke outcome may provide new biomarkers or therapeutic approaches for ischemic stroke. Here, we prospectively recruited 180 first-ever AIS patients within 72 hrs after stroke onset. We used the National Institutes of Health Stroke Scale (NIHSS) to quantify stroke severity and modified Rankin scale (mRS) to assess the 3-month outcome for AIS patients. Initially, we screened 35 cytokines, chemokines, and growth factors in sera from 75 AIS patients and control subjects. Cytokines that were of interest were further investigated in the 180 AIS patients and 14 heathy controls. We found that IL-1RA, IL-1β, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-13, IL-15, EGF, G-CSF, Flt-3L, GM-CSF and Fractalkine levels were significantly decreased in severe stroke patients. In particular, IL-1β, IL-4, IL-5, IL-7, IL-9, IL-10, IL-15, G-CSF and GM-CSF were significantly reduced in AIS patients with poor outcome, compared to those with good prognosis. IL-6 was notably higher in the poor outcome group. Only IL-9 level decreased in the large infarct volume group. After adjusting for confounders, we found that IL-5 was an independent protective factor for prognosis in AIS patients with an adjusted OR of 0.042 (P = 0.007), whereas IL-6 was an independent risk predictor for AIS patients with an adjusted OR of 1.293 (P = 0.003). Our study suggests the levels of serum cytokines are related to stroke severity, short-term prognosis and cerebral infarct volume in AIS patients.

Protein array profiling of circulating angiogenesis-related factors during bevacizumab containing treatment in metastatic colorectal cancer.

BackgroundProlonged angiogenesis inhibition may improve treatment outcome in metastatic colorectal cancer (mCRC) patients. However, due to the complexity of the angiogenic pathways there is a lack of valid predictive biomarkers for anti-angiogenic agents. Here, we describe and optimize a procedure for simultaneous dynamic profiling of multiple angiogenesis related proteins in patient serum to explore associations with the response and acquired resistance to anti-angiogenic therapy.Materials and methodsPatients (n=22) were selected from a clinical trial investigating maintenance treatment with bevacizumab alone after response to induction chemotherapy + bevacizumab in mCRC. Serum samples were analysed for 55 unique angiogenesis related proteins using a commercial proteome profiler array and a publicly available image analysis program for quantification. Samples were collected at baseline before induction treatment start, at start of maintenance treatment, and at end of treatment after tumour progression.Main results and conclusionFor eight proteins, the antibody array signals were below detection range in all patient samples. None of the proteins showed levels at baseline or at start of maintenance with strong evidence for correlation to time to progression (lowest nominal p-value 0.03). The dynamic ranges of protein levels measured during the induction treatment period and during the maintenance period were analysed separately for time trends. Evidence for changing trends (up/down) in the levels of MMP-8, TIMP-4 and EGF was observed both during response to induction treatment and at progressive disease, respectively. For three of the proteins (IL-8, Activin A and IGFBP-2), weak evidence for correlation between increasing protein levels during induction with chemotherapy and bevacizumab and time to progression was observed.In conclusion, semi-quantitative profiling of angiogenesis related proteins in patient serum may be a versatile tool to screen for protein patterns aiming at identifying resistance mechanisms of anti-angiogenic treatment in patients with mCRC.

Epidermal Growth Factor Rejuvenates Aging Hematopoietic Stem Cells

Aging hematopoietic stem cells display distinct abnormalities such as myeloid skewing, decreased repopulating capacity, and leukemia predisposition. Radiation exposure potentiates aging of the hematopoietic system, as evidenced by premature and persistent senescence of HSCs in mice exposed to moderate dose irradiation (Blood, 2014. 123(20): p. 3105-15). Our laboratory demonstrated that EGF promotes hematopoietic stem cell (HSC) regeneration after radiation injury (Nat Med, 2013. 19(3): p. 295-304). Based on these results, we hypothesized that EGF and EGFR signaling may have rejuvenating effects on aging HSCs.For comparisons of young and aged mice, we utilized 2-4 month old mice and >18-24 month old female C57BL/6 mice, respectively. We discovered that aged C57BL/6 mice have decreased levels of EGF in the peripheral blood compared to young mice (p= 0.02) and decreased expression of EGFR on bone marrow (BM) ckit+sca-1+lin- (KSL) stem/progenitor cells (p= 0.03). BM KSL cells from aged mice displayed increased DNA damage in culture with thrombopoietin, SCF and Flt-3 ligand (TSF) compared to young KSL cells (p< 0.0001). Aged BM KSL cells treated with 100 ng/ml EGF had decreased DNA damage compared to aged BM KSL cells treated with TSF only (p< 0.01) Treatment with EGF for 7 days increased the colony forming capacity of aged BM KSL cells (p< 0.01). Importantly, competitive repopulation assays revealed that 7 day in vitro treatment of aged BM KSL cells with EGF significantly increased primary donor cell engraftment (p< 0.01) at 16 weeks compared to aged BM KSL cells cultured with TSF alone.Based on these in vitro observations, we next tested whether systemic administration of a recombinant, pegylated EGF (pEGF), 20 mcg three times weekly for 4 weeks could alter the hematopoietic characteristics of aged C57BL/6 mice. pEGF treatment decreased BM myeloid skewing (p< 0.01) and increased CD3 T cell content (p< 0.01) in aged mice compared to saline-injected controls. Furthermore, competitive repopulation assays revealed that pEGF treatment of aged mice caused a significant increase in functional HSCs capable of competitive engraftment in recipient congenic mice. Total primary donor cell (p< 0.01), myeloid cell (p< 0.01), T cell (p< 0.01) and B cell (p= 0.01) engraftment were significantly increased in recipient mice transplanted with BM cells from aged, pEGF-treated donors compared to recipients of aged, saline-treated donors at 20 weeks. Additionally, secondary transplantation also resulted in increased donor engraftment (p< 0.01) and increased donor myeloid engraftment (p< 0.05) at 12 weeks. These results suggested that systemic administration of pEGF rejuvenates the repopulating ability of aged HSCs.We next sought to determine if deficiency in EGFR signaling could accelerate hematopoietic aging in mice. For this purpose, we utilized a doxycycline-inducible, hematopoietic cell specific EGFR dominant negative mutant model (SCL-tTA;EGFR-DN mice). Interestingly, adult (13 month old) SCL-tTA;EGFR-DN mice displayed increased myeloid skewing in the peripheral blood (p< 0.05) and BM cells from SCL-tTA;EGFR-DN mice generated significantly decreased colony forming cells, compared to age-matched, EGFR-expressing mice (p< 0.01). Mechanistically, BM KSL cells from 13 month old SCL-tTA;EGFR-DN mice demonstrated increased senescence (p< 0.01) and increased expression of p16 (p< 0.05) compared to young mice. In contrast, BM KSL cells from aged C57BL/6 mice that were treated with 100 ng/ml EGF (plus TSF) for 7 days displayed decreased senescence and decreased p16 expression compared to BM KSL cells from aged mice treated with TSF alone. Taken together, these studies suggest that EGF/EGFR signaling declines with age and that reactivation of EGFR signaling via EGF treatment can ameliorate clinically relevant features of hematopoietic aging, including HSC self-renewal capacity. DisclosuresNo relevant conflicts of interest to declare.

N-O Reduction and ROS-Mediated AKT/FOXO1 and AKT/P53 Pathways Are Involved in Growth Promotion and Cytotoxicity of Cyadox.

Cyadox is a novel derivative of quinoxaline-1,4-dioxides (QdNOs) with the potential to be developed as a feed additive. However, the pharmacological and toxicological bioactive molecules of cyadox and the molecular mechanism of its pharmacological and toxic actions remain unclear. In the present study, cyadox and its main metabolites of cy1, cy4, cy6, and cy12 were selected; the growth promotion characteristic was indicated by the mRNA level of EGF; and the cytotoxicity of cyadox was determined by methylthiazol tetrazolium bromide (MTT) assay, lactate dehydrogenase (LDH) release, and Annexin V-FITC/PI apoptosis detection kit with flow cytometry. The intracellular ROS, cyclin D1, and Akt/P53/FOXO1 signaling pathway were also investigated. Our data suggested that cyadox showed relatively higher activity than its metabolites, and the ROS was generated from N-O reduction of cyadox. Moreover, cyadox (2 μM) activated the Akt and increased the EGF, cyclin D1, and FOXO1 expression levels. Cyadox (100 μM) induced cytotoxicity in L02 cells in a concentration- and time-dependent manner. Additionally, the activated P53 pathway, hyperactivated Akt, and apoptosis were found in L02 cells after incubation with 100 μM cyadox. Our data demonstrated that Akt promoted cell survival when it was mildly activated by cyadox at 2 μM, and Akt leads to apoptosis when it was severely activated by cyadox at 100 μM. Thus, the present study revealed that N-O reduction of cyadox and ROS-mediated AKT/FOXO1 and AKT/P53 pathways were involved in growth promotion and cytotoxicity of cyadox.

Tunable activation of therapeutic platelet-rich plasma by pulse electric field: Differential effects on clot formation, growth factor release, and platelet morphology.

BackgroundActivation of platelet-rich plasma (PRP) by pulse electric field (PEF) releases growth factors which promote wound healing (e.g., PDGF, VEGF for granulation, EGF for epithelialization).AimsTo determine after PEF activation of therapeutic PRP: 1) platelet gel strength; 2) profile of released growth factors; 3) alpha- and T-granule release; 4) platelet morphology.MethodsConcentrated normal donor PRP was activated by 5 μsec (long) monopolar pulse, ~4000 V/cm (PEF A) or 150 nsec (short) bipolar pulse, ~3000 V/cm (PEF B) in the presence of 2.5 mM (low) or 20 mM (high) added CaCl2. Clot formation was evaluated by thromboelastography (TEG). Surface exposure of alpha granule (P-selectin) and T-granule (TLR9 and protein disulfide isomerase [PDI]) markers were assessed by flow cytometry. Factors in supernatants of activated PRP were measured by ELISA. Platelet morphology was evaluated by transmission electron microscopy (TEM).ResultsTime to initial clot formation was shorter with thrombin (<1 min) than with PEF A and B (4.4–8.7 min) but clot strength (elastic modulus, derived from TEG maximum amplitude) was greater with PEF B than with either thrombin or PEF A (p<0.05). Supernatants of PRP activated with PEF A had higher EGF levels than supernatants from all other conditions. In contrast, levels of PF4, PDGF, and VEGF in supernatants were not significantly different after PEF A, PEF B, and thrombin activation. T-granule markers (TLR9 and PDI) were higher after thrombin than after PEF A or B with low or high CaCl2. By TEM, platelets in PEF-treated samples retained a subset of granules suggesting regulated granule release.ConclusionPulse length and polarity can be modulated to produce therapeutic platelet gels as strong or stronger than those produced by thrombin, and this is tunable to produce growth factor profiles enhanced in specific factors important for different stages of wound healing.

The Antimicrobial Peptide Nal-P-113 Exerts a Reparative Effect by Promoting Cell Proliferation, Migration, and Cell Cycle Progression.

Objective The primary purpose of this study was to evaluate the reparative efficacy of a novel antimicrobial peptide, Nal-P-113, in shortening the healing time of oral mucosal ulcers by promoting cell proliferation and migration and accelerating the cell cycle. Methods Cell counting kit-8 (CCK-8) and wound-healing assays were used to evaluate the proliferation and migration of human immortalized oral epithelial cells (HIOECs). The cell cycle distribution of HIOECs was analyzed by flow cytometry. Additionally, the RNA levels of EGF, FGF-2, and TGF-β1 of HIOECs were assessed by real-time PCR. Rats were divided into three groups randomly: (a) blank control group; (b) 20 μg/mL Nal-P-113; and (c) 10 ng/mL rhEGF. An oral mucosal ulcer was induced in every rat by the application of 30% acetic acid. An immunohistochemical assay was used to assess the expression of EGF, FGF-2, and TGF-β1 in the rat oral mucosa. Results In the CCK-8 assay, the optical density values in the Nal-P-113 and rhEGF groups were found to be significantly higher than that in the blank control group. In addition, the scratch areas in the Nal-P-113 and rhEGF groups were found to be significantly smaller (P<0.05). Cell cycle analysis showed that Nal-P-113 accelerated the entry of HIOECs into the S phase and expedited their cell cycles. The RT-PCR results suggested that Nal-P-113 upregulated the RNA levels of EGF and FGF-2 but downregulated that of TGF-β1 at 24 h and 48 h. Lastly, the immunohistochemical assay verified that Nal-P-113 changed the expression of the above cytokines in rat mucosal ulcers. Conclusion Nal-P-113 promoted the repair of oral mucosal ulcers by increasing the EGF and FGF-2 expression and decreasing that of TGF-β1 in HIOECs, accelerating their proliferation and cell cycle progression. The application of Nal-P-113 might serve as an effective therapeutic approach for recurrent aphthous stomatitis.

Effects of oral administration of peptides with low molecular weight from Alaska Pollock (Theragra chalcogramma) on cutaneous wound healing

Protein supplements play an important role in wound healing. The effect of collagen peptide (PCP) and flesh protein peptide (FPP) from Alaska Pollock on wound healing was investigated. The molecular weight distributions of PCP and FPP were in a range of 430–1000 Da and 100–1000 Da, respectively. Compared to vehicle group, the wound healing activities in PCP and FPP were significantly enhanced, which could be corroborated by the increase of wound healing rate, hydroxylproline content, and tensile strength in rat wound model (P < 0.05). H&E showed a near-normal epidermis structure of PCP and FPP on day 12, but vehicle group displayed poor re-epithelialization. PCP and FPP increased the levels of bFGF, EGF, TGF-β1 and TβRII (P < 0.05), and decreased the content of Smad7 in TGF/Smad signaling pathway. Therefore, FPP and PCP can be used as effective ingredients in the treatment of wound healing.

Valproate augments Niraparib killing of tumor cells

ABSTRACTPARP1 inhibitors are approved therapeutic agents in ovarian carcinomas, and have clinical activity in some breast cancers. As a single agent, niraparib killed ovarian and mammary tumor cells via an ATM-AMPK-ULK1 pathway which resulted in mTOR inactivation and the formation of autophagosomes, temporally followed by autolysosome formation. In parallel, niraparib activated a CD95-FADD-caspase 8 pathway, and collectively these signals caused tumor cell death that was suppressed by knock down of Beclin1, ATG5, CD95, FADD or AIF; or by expression of c-FLIP-s, BCL-XL or dominant negative caspase 9. The HDAC inhibitors AR42 and sodium valproate enhanced niraparib lethality in a greater than additive fashion. HDAC inhibitors enhanced niraparib lethality by increasing activation of the ATM-AMPK-ULK1-autophagy and CD95-FADD-caspase 8 pathways. Knock down of eIF2α, ATM, AMPKα, ULK1, Beclin1 or ATG5 reduced tumor cell killing by the niraparib plus HDAC inhibitor combination. Blockade of either caspase 9 function or that of cathepsin B partially prevented cell death. As a single agent niraparib delayed tumor growth, but did not significantly alter the tumor control rate. Tumors previously exposed to niraparib had activated the ERK1/2 and AKT-mTOR pathways that correlated with increased plasma levels of IL-8, MIF, EGF, uPA and IL-12. Collectively our findings argue that the addition of HDAC inhibitors to niraparib enhances the anti-cancer activity of the PARP1 inhibitor niraparib.

Burn wounds in the young versus the aged patient display differential immunological responses

BackgroundIndividuals in the geriatric age range are more prone than younger individuals to convert their partial thickness thermal burns into full thickness injuries. We hypothesized that this often observed clinical phenomenon is strongly related to differential local injury responses mediated by the immune system. Materials & methodsSkin samples from areas with partial thickness thermal burns were obtained during routine excision and grafting procedures between post burn days 2–6. Tissue samples were grouped by age ranges with young patients defined as <30 years of age or aged patients defined as >65. Formalin fixed samples were used to confirm depth of burn injury and companion sections were homogenized for multiplex analysis using a Luminex platform. Immunohistochemical staining was used to quantify total macrophage numbers as well as the M1 and M2 subpopulations. ResultsOur analysis includes samples derived from 11 young subjects (mean age=23) and 3 aged subjects (mean age=79.2). Our initial survey of analytes examined 31 cytokines/chemokines. Twelve were excluded from consideration as they were present in concentrations either above or below the optimal detection range. Two analytes emerged as candidate molecules with significant differences between the young and the aged patient responses to burn injury. EGF levels were on average 21.69pg/ml in young vs 14.87pg/ml in aged (p=0.032). RANTES/CCL5 levels were on average 14.86pg/ml in young vs 4.26pg/ml in aged (p=0.026). Elevated macrophage numbers were present within wounds of younger patients compared to the old (p<0.01), with a higher concentration of the M1 type in the elderly (p>0.05). ConclusionOur study has identified at least 2 well known cytokines, CCL5 (RANTES) and EGF, which are differentially regulated in response to burn injury by young versus aged burn victims. Evidence suggests that a proinflammatory environment can explain the high conversion rate from partial to full thickness burns. Our data suggest the need for future studies at the point of injury (cutaneous targets) that may be modulated by post burn release of cytokines/chemokines.

Inflammatory patterns in Takotsubo cardiomyopathy and acute coronary syndrome: A propensity score matched analysis

Background and aimsSystemic inflammatory activation can be observed in both Takotsubo cardiomyopathy (TTC) and acute coronary syndrome (ACS). The aim of this study was therefore to compare circulating cytokine levels during the acute and subacute phase of TTC and ACS. MethodsOne hundred thirty-six consecutive patients were enrolled in the study; after a propensity score matching, 32 TTC patients were compared with 32 subjects with ACS. Clinical baseline features and circulating levels of interleukin(IL)-1β, IL-1α, IL-2, IL-4, IL-6, IL-8, IL-10, IFN-γ, MCP1, EGF, VEGF, TNFα were obtained at admission (t0) and after 120 h (t1). ResultsAt t0, several circulating IL levels were higher in subjects with TTC (IL-2 2 vs. 0.5 pg/ml, IL-4 1.5 vs. 0.82 pg/ml, IL-10 3.34 vs. 1.62 pg/ml, TNF-α 5 vs. 2.3 pg/ml, IFN-γ 0.92 vs. 0.32 pg/ml, EGF 84.8 vs. 10.7 pg/ml, p < 0.05 in all cases), while IL-6 levels were higher in patients with ACS (25.4 vs. 12.4 pg/ml p = 0.03).At t1, IL-2 and EGF levels were still higher in patients with TTC vs. those with ACS (IL-2 4.6 vs. 0.72 pg/ml, p = 0.01; EGF 36.3 vs. 18.5 pg/ml, p = 0.03), while IL-6 serum levels were higher in ACS patients (19.6 vs. 7.35 pg/ml, p = 0.02). ConclusionsDifferent inflammatory patterns can be observed during the acute and subacute phase of TTC when compared to ACS. Increased levels of anti-inflammatory interleukins can be found during the acute phase of TTC while ACS is featured by higher levels of IL-6 during the acute and sub-acute phase.

Do high VEGFA and EGF levels in blood indicate liver metastases only?

e24007Background: Liver metastasis is a common sign of tumor progression. Growth factors are tissue malignancy markers, while their dynamics in the blood of cancer patients is poorly studied. The p...

Elevated EGF Levels in the Blood Serum of Dogs with Periodontal Diseases and Oral Tumours.

Paradontopathy and neoplasms of the oral cavity represent one of the greatest challenges in human and animal dentistry. EGF plays a key role in maintaining the integrity and proper rate of cell proliferation in normal oral epithelium. The aim of the present study was to study serum levels of EGF in dogs diagnosed with periodontal diseases and oral cavity tumours. The samples comprised of cancerous tissue sections and serum obtained from dogs of various breeds, aged between 5-13 years. Serum EGF concentrations were measured by an immunoenzymatic method. The median for EGF concentration in serum of dogs suffered from severe periodontal diseases was greater when compared to the control group. EGF concentration in dogs with malignant tumours was significantly higher than in those with non-malignant growths. A positive correlation between EGF concentration and tumour size was also observed. EGF level in dogs diagnosed with benign tumours was comparable to the control group. The blood serum level of EGF increases significantly in patients with malignant oral tumours and advanced periodontal disease. In malignant tumours, the high level of EGF correlates with the size and invasiveness of the neoplasm.

Development of c‐Cbl Antagonists to Promote Corneal Epithelial Wound Healing

Ligand mediated desensitization is a key negative regulator of cell surface receptors. In the human cornea, epidermal growth factor receptor (EGFR) desensitization, due to the relatively high levels of endogenous EGF in the tear fluid, limits the repair and restoration of the corneal epithelium. The E3 ubiquitin ligase, c‐Cbl is a critical modulator of EGFR desensitization, as it mediates the ubiquitylation of the activated receptor and targets it to the lysosome for degradation. Ultimately, this terminates receptor signaling. We have previously demonstrated that knock down of c‐Cbl results in an enhancement of EGFR‐mediated cell migration that is necessary for healing of corneal epithelial wounds. The goal of these experiments was to identify novel compounds that antagonize c‐Cbl activity and could be developed to enhance EGFR‐mediated corneal epithelial wound healing. Using the published crystal structures of the EGFR:c‐Cbl complex, we identified the point of interaction between the two proteins. We subsequently performed an in silico screen of ~25,000,000 compounds (from the ZINC15 library) to identify compounds that are predicted to antagonize c‐Cbl binding to and ubiquitylation of the EGFR. The top 50 candidates were screened using a biochemical assay for their ability to bind c‐Cbl using Differential Scanning Fluorimetry (Thermofluor™). From that screen, six compounds were identified as c‐Cbl binders and were tested in immortalized corneal epithelial cells to determine which compounds could inhibit c‐Cbl function. Two of the six compounds blocked EGF‐mediated EGFR ubiquitylation, albeit at relatively high concentrations (IC50 were ~50 μM and ~5 μM). Further, these compounds exhibited some cytotoxicity at 100 μM. These compounds represent two lead compounds that can be chemically optimized to enhance their affinity for c‐Cbl and become more potent inhibitors of c‐Cbl activity. The long‐term goal is to use these compounds as therapeutic agents to promote the restoration of damaged corneal epithelium, to minimize patient discomfort, reduce ocular infections, and minimize the likelihood of blindness.Support or Funding InformationResearch was supported by NIH/NEI grant EY027032.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Reduction of soluble dipeptidyl peptidase 4 levels in plasma of patients infected with Middle East respiratory syndrome coronavirus

Dipeptidyl peptidase 4 (DPP4) is a receptor for MERS-CoV. The soluble form of DPP4 (sDPP4) circulates systematically and can competitively inhibit MERS-CoV entry into host cells. Here, we measured the concentration of sDPP4 in the plasma and sputa of 14 MERS-CoV-infected patients of various degrees of disease severity. The concentration of sDPP4 in the plasma of MERS patients (474.76 ± 108.06 ng/ml) was significantly lower than those of healthy controls (703.42 ± 169.96 ng/ml), but there were no significant differences among the patient groups. Interestingly, plasma levels of IL-10 and EGF were negatively and positively correlated with sDPP4 concentrations, respectively. The sDPP4 levels in sputa were less than 300 ng/ml. Viral infection was inhibited by 50% in the presence of more than 8000 ng/ml of sDPP4. Therefore, sDPP4 levels in the plasma of MERS patients are significantly reduced below the threshold needed to exert an antiviral effect against MERS-CoV infection.

Is there association between characteristics of gastric tumor growth and activation of different growth factor systems?

29 Background: Signet-ring cell carcinoma of the stomach (SRCC) is significantly associated with poor prognosis and low treatment effectiveness, compared to adenocarcinoma of the stomach (AC). One of the causes of SRCC aggressive course can be its diffuse invasive growth pattern. The ability of tumor cells to synthesize and/or capture growth factors ensures the independent tumor development. VEGF, EGF, IGFI and EGFII facilitate neoangiogenesis and regulation of malignant processes in paracrine/autocrine mechanisms. The purpose of the study was to find the difference between local levels of VEGF, EGF, IGFI and IGFII in tumor and intact tissues in AC and SRCC. Methods: Levels of VEGF, EGF, EGFR, IGF-I and IGFII were determined by ELISA in tumor and intact gastric tissues of 15 AC patients and 10 SRCC patients (T2-3N0-2M0, G2-3), mean age 62.8±2.38 years. Results: Levels of IGF-I, IGF-II and VEGF in intact gastric tissues were similar in AC and SRCC. EGF and EGFR levels in AC were 2.9 and 1.5 higher than in SRCC. Levels of VEGF in AC tumor tissues, compared to intact gastric tissues, increased by 2.3 times, EGF and EGFR did not change, and IGFI and IGFII decreased by 2.1 and 1.8 times, respectively. Tumor tissues of SRCC patients showed the increased levels of EGF (by 2.9 times) and EGFR (by 1.8 times); VEGF and IGF were similar to the levels in intact tissues. SRCC was characterized by the activation of systems of EGF and IGF, but not VEGF. AC demonstrated the stimulation of VEGFA neoangiogenesis. Conclusions: The activation of systems of EGF and IGF in SRCC could be associated with some characteristics of the diffuse growth pattern and the tumor origin from neuroendocrine cells which facilitates the autocrine/paracrine regulation of the invasive growth. AC requires neoangiogenesis which is realized primarily by the VEGF system.