Human Epididymis Protein 4 Concentrations Research Articles

Urinary-based detection of MSL, HE4 and CA125 as an additional dimension for predictive and prognostic modelling in ovarian cancer

ObjectivesWe have recently described a predictive/prognostic model for ovarian cancer, exploiting commonly available clinico-pathological parameters and the ovarian serum biomarkers mesothelin (MSL), human epididymis protein 4 (HE4) and cancer-antigen 125 (CA125). Considering urine as a prototype non-invasive sample, we investigated whether serum levels of these biomarkers are mirrored in urine and compared their clinical relevance in matched serum vs. urine samples.MethodsMSL, HE4 and CA125 were quantified in urinary (n=172) and matched serum samples (n=188) from ovarian cancer patients (n=192) using the Lumipulse® G chemiluminescent enzyme immunoassay (Fujirebio).ResultsWhile absolute concentrations of MSL or CA125 were higher in serum than in matched urine samples, HE4 concentrations were considerably higher in urine than in serum. Nonetheless, the levels of all three biomarkers strongly correlated between matched serum vs. urine samples and were unrelated to BRCA1/2 mutational status. Consequently, prediction of surgical outcome or relapse/death by MSL, HE4 or CA125 was similarly efficient among urinary- vs. serum-based detection. HE4 provided the highest capacity to predict surgical outcome or relapse/death among both body fluids (urine: AUC=0.854; serum: AUC=0.750, respectively). All clinically relevant findings regarding the investigated urinary biomarkers were equally reproducible among raw vs. creatinine-normalized datasets, suggesting that normalization may have subordinate priority for urine-based analysis of these biomarkers.ConclusionWe report that the capacity of MSL, HE4 and CA125 to predict surgical outcome and relapse/death is equivalent between serum vs. urine-based detection. Urinary biomarkers, in particular HE4, may provide an additional dimension for prognostic modeling in ovarian cancer.

Cooperative amplification of Prussian blue as a signal indicator and functionalized metal-organic framework-based electrochemical biosensor for an ultrasensitive HE4 assay

Human epididymis protein 4 (HE4), a diagnostic biomarker of ovarian cancer, is crucial for monitoring the early stage of the disease. Hence, it is highly important to develop simple, inexpensive, and user-friendly biosensors for sensitive and quantitative HE4 assays. Herein, a new sandwich-type electrochemical immunosensor based on Prussian blue (PB) as a signal indicator and functionalized metal-organic framework nanocompositesas efficient signal amplifiers was fabricated for quantitative analysis of HE4. In principle, ketjen black (KB) and AuNPs modified on TiMOF (TiMOF-KB@AuNPs) could accelerate electron transfer on the electrode surface and act as a matrix for the immobilization of antibodies via cross-linking to improve the determination sensitivity. The PB that covalently binds to labeled antibodies endows the biosensors with intense electrochemical signals. Furthermore, the concentration of HE4 could be indirectly detected by monitoring the electroactivity of PB. Benefiting from the high signal amplification ability of the PB and MOF nanocomposites, this strategy displayed a wide linear range (0.1–80 ng mL−1) and a lower detection limit (0.02 ng mL−1). Hence, this study demonstrated great promise for application in clinical ovarian cancer diagnosis and treatment, and provided a new platform for detecting other cancer biomarkers.

Urinary-based detection of MSL, HE4, and CA125 as an additional dimension for predictive and prognostic modelling in ovarian cancer.

e17567 Background: We have recently described a predictive/prognostic model for ovarian cancer, exploiting commonly available clinico-pathological parameters and the ovarian serum biomarkers mesothelin (MSL), human epididymis protein 4 (HE4) and cancer-antigen 125 (CA125). Considering urine as a prototype non-invasive sample, we investigated whether serum levels of these biomarkers are mirrored in urine and compared their clinical relevance in matched serum vs. urine samples. Methods: MSL, HE4 and CA125 were quantified by Lumipulse G chemiluminescent enzyme immunoassay (Fujirebio). In total, this study was based on 1080 biomarker detection runs from 188 serum and 172 urine samples from 192 ovarian cancer patients. Results: While absolute concentrations of MSL and CA125 were higher in serum than in urine, serum HE4 concentrations were considerably lower than urinary HE4 concentrations. Nonetheless, relative intra-patient levels of all three biomarkers strongly correlated between matched serum vs. urinary samples and were unrelated to BRCA1/2 mutational status. Consequently, prediction of surgical outcome or relapse/death by MSL, HE4 or CA125 was similarly efficient among urinary- vs. serum-based detection. HE4 provided the highest capacity in predicting surgical outcome and relapse/death among both body fluid types (urinary HE4: AUC=0.854; AUC=0.750, respectively). All clinically relevant findings of urinary MSL, HE4 and CA125 were reproducible among raw vs. creatinine-normalized datasets, suggesting that normalization may have subordinate priority for urine-based analysis. Conclusions: We report that the capacity of MSL, HE4 and CA125 for predicting surgical outcome and relapse/death is equivalent between serum vs. urine-based detection. Urinary biomarkers, in particular HE4, may provide an additional dimension for prognostic modeling in ovarian cancer.

Elevated level of serum human epididymis protein 4 (HE4) predicts disease severity and mortality in COVID-19 pneumonia

BackgroundWe retrospectively analyzed serum level of human epididymis protein 4 (HE4) as a pulmonary inflammatory biomarker in patients with COVID-19 pneumonia in association with disease severity and outcome.MethodsNinety-nine (40 critically ill, 40 severe and 19 mild) COVID-19 patients and as controls 25 age- and sex-matched non-COVID-19 bacterial sepsis subjects were included. Serum HE4 was measured by an immunoassay (Architect® i1000SR, Abbott) in the baseline samples of all study participants obtained at intensive care unit (ICU) admission or during outpatient clinic visit and follow-up sera were available in case of 30 COVID-19 subjects with life-threating conditions. Associations were studied between serum HE4, routinely available laboratory parameters, clinical characteristics, and disease progression.ResultsBaseline HE4 level was significantly higher (P < 0.0001) in critically ill (524.7 [300.1–1153.0] pmol/L) than severe COVID-19 subjects (157.4 [85.2-336.9] pmol/L) and in mild SARS-CoV-2 infection (46.7 [39.1–57.2] pmol/L). Similarly increased HE4 concentrations were found in bacterial sepsis (1118.0 [418.3–1953.0] pmol/L, P = 0.056) compared to critically ill COVID-19 individuals. Serum HE4 levels significantly correlated with age, SOFA-score, inflammation-dependent biomarkers, and the degree of lung manifestation evaluated by chest CT examination in ICU COVID-19 individuals. Based on ROC-AUC curve analysis, baseline HE4 independently indicated the severity of COVID-19 with an AUC value of 0.816 (95% CI [0.723–0.908]; P < 0.0001), while binary logistic regression test found HE4 as an independent prognostic parameter for death (OR: 10.618 [2.331–48.354]; P = 0.002). Furthermore, COVID-19 non-survivors showed much higher baseline HE4 levels without a substantial change under treatment vs. survivors (P < 0.0001). Finally, pre-treatment HE4 level of ≥ 331.7 pmol/L effectively predicted a larger risk for mortality (Log-Rank P < 0.0001) due to severe COVID-19 pneumonia.ConclusionElevated serum HE4 level at ICU admission highly correlates with COVID-19 severity and predicts disease outcome.

P174 Human epididymis protein 4 (HE4) plasma concentrations correlate with the improvement of ppFEV1 in response to LUM/IVA therapy in people with cystic fibrosis homozygous for p.Phe508del-CFTR

We have previously documented that elevated plasma HE4 concentrations decreased in people with CF (pwCF) bearing p.Gly551Asp-CFTR mutation in response to ivacaftor (IVA), and these concentrations were inversely correlated with ppFEV1. Although the PROSPECT study has evaluated the effectiveness of lumacaftor (LUM)/IVA in pwCF homozygous for p.Phe508del-CFTR variant, plasma biomarkers have not been used to monitor treatment efficacy.

Human epididymis protein 4 (HE4) plasma concentration inversely correlates with the improvement of cystic fibrosis lung disease in p.Phe508del-CFTR homozygous cases treated with the CFTR modulator lumacaftor/ivacaftor combination

BackgroundWe previously documented that elevated HE4 plasma concentration decreased in people with CF (pwCF) bearing the p.Gly551Asp-CFTR variant in response to CFTR modulator (CFTRm) ivacaftor (IVA), and this level was inversely correlated with the FEV1% predicted values (ppFEV1). Although the effectiveness of lumacaftor (LUM)/IVA in pwCF homozygous for the p.Phe508del-CFTR variant has been evaluated, plasma biomarkers were not used to monitor treatment efficacy thus far. MethodsPlasma HE4 concentration was examined in 68 pwCF drawn from the PROSPECT study who were homozygous for the p.Phe508del-CFTR variant before treatment and at 1, 3, 6 and 12 months after administration of LUM/IVA therapy. Plasma HE4 was correlated with ppFEV1 using their absolute and delta values. The discriminatory power of delta HE4 was evaluated for the detection of lung function improvements based on ROC-AUC analysis and multiple regression test. ResultsHE4 plasma concentration was significantly reduced below baseline following LUM/IVA administration during the entire study period. The mean change of ppFEV1 was 2.6% (95% CI, 0.6 to 4.5) by 6 months of therapy in this sub-cohort. A significant inverse correlation between delta values of HE4 and ppFEV1 was observed especially in children with CF (r=-0.7053; p<0.0001). Delta HE4 predicted a 2.6% mean change in ppFEV1 (AUC: 0.7898 [95% CI 0.6823–0.8972]; P < 0.0001) at a cut-off value of -10.7 pmol/L. Moreover, delta HE4 independently represented the likelihood of being a responder with ≥ 5% delta ppFEV1 at 6 months (OR: 0.89, 95% CI: 0.82–0.95; P = 0.001). ConclusionsPlasma HE4 level negatively correlates with lung function improvement assessed by ppFEV1 in pwCF undergoing LUM/IVA CFTRm treatment.

Label-free electrochemical bioplatform based on Au-modified magnetic Fe3O4/α-Fe2O3 hetero-nanorods for sensitive quantification of ovarian cancer tumor marker

Due to challenges in the ultrasensitive detection of human epididymis protein 4 (HE4), a vital tumor marker for the early diagnosis of ovarian cancer (OC), it is necessary to establish efficient methods for HE4 determination. Recently, electrochemical bioplatforms combined with nanomaterials have shown outstanding performance in the analysis of biological molecules. The surface modification of conductive materials is essential for signal amplification and detection sensitivity. Herein, we developed a novel label-free HE4 electrochemical bioplatform by introducing magnetic Fe3O4/α-Fe2O3 hetero-nanorods (Fe3O4/α-Fe2O3 NRs) as a substrate for signal amplification and DNA aptamer (Apt) probe immobilization in combination with magnetic self-assembly and magnetic separation. Differential pulse voltammetry (DPV) results verified that the platform had excellent specificity for HE4 under optimized conditions and was positively linearly correlated with the logarithm of HE4 concentration (R2 = 0.996) with a linear range of 0.1 pg mL−1 to 1 ng mL−1 and detection limit (LOD) of 27.5 fg mL−1. Moreover, the bioplatform demonstrated promising results for the detection of HE4 in spiked serum samples.

Serum Human Epididymis Protein 4 as a Prognostic Predictor of New-Onset Heart Failure among Women after Acute Coronary Syndrome: A Single-Center Retrospective Study

Introduction: Little is known about the prognostic factors among women with acute coronary syndrome (ACS), partly due to the small number of women included in heart failure (HF) clinical trials. Human epididymis protein 4 (HE4) has been proven to be a new biomarker for acute and chronic HF over the years. We hypothesize that HE4 could be a promising predictor. Methods: This retrospective study analyzed data from Zhejiang Provincial People’s Hospital. This study included 302 female patients with ACS between January 1, 2021, and December 1, 2021. The primary outcome was new-onset HF after ACS during the 12-month follow-up period. We used a logistic regression model to evaluate the association between serum HE4 levels and the incidence of HF. Serum HE4 levels were measured at baseline (within 24 h after admission). Results: Of the 302 female patients, 70 (23.2%) developed new-onset HF within 12 months. Serum HE4 levels in patients with adverse events were significantly higher than those in patients without events (8.9 [7.3–11.5] pmol/dL versus 5.9 [5.0–6.8] pmol/dL, p < 0.001). The levels of HE4, troponin I peak, left ventricular ejection fraction (LVEF), and estimated glomerular filtration rate (eGFR) were validated as independent predictors, with HE4 being the best laboratory predictor (area under the curve, 0.863; 95% confidence interval, 0.817–0.909). Serum HE4 concentrations of >6.93 pmol/dL distinguished patients at risk of HF with 82.9% sensitivity and 78.0% specificity (maximum Youden index J, 0.609). Moreover, HE4 levels were associated with an increased risk of HF. Discussion: We found a strong relationship between HE4 and the occurrence of HF after ACS among women, which might help identify patients at high risk of HF for whom close or intense management should be mandatory.

The use of CA125, human epididymis protein 4 (HE4), risk of ovarian malignancy algorithm (ROMA), risk of malignancy index (RMI) and subjective assessment (SA) in preoperative diagnosing of ovarian tumors.

To compare utility of CA125, human epididymis protein 4 (HE4), risk of ovarian malignancy algorithm (ROMA), risk of malignancy index (RMI) and subjective assessment (SA) in preoperative diagnosis of ovarian tumors. Research was conducted among 456 patients qualified for surgery due to ovarian tumor. Preoperatively, CA125 and HE4 serum levels were estimated, and transvaginal ultrasound was performed. ROMA and RMI values and SA qualifications were obtained. Results were compared with pathomorphological findings. Receiver Operating Characteristic (ROC)-Area Under Curve (AUC) values for CA125, HE4, ROMA, RMI and SA in preoperative diagnosis of malignant lesions were 0.819, 0.909, 0.911, 0.895 and 0.895, respectively. Combinations of biochemical and sonographic methods increased sensitivity in diagnosis of ovarian tumors. Combinations utilizing serum HE4 concentrations were most useful. CA125, HE4, ROMA, RMI and SA proved to be useful in preoperative diagnosis of ovarian tumors. HE4 and ROMA occurred to be the most useful. Ultrasonographic methods are considerably useful in diagnosis of ovarian tumors. RMI and SA present similar overall diagnostic value.

Serum human epididymis secretory protein 4 correlates with sepsis-associated acute respiratory distress syndrome and 28-day mortality in critically ill patients.

Acute respiratory distress syndrome (ARDS) is a severe disease with high mortality, and its primary cause is sepsis. The aim of this study was to detect and evaluate the role of Human epididymis protein 4 (HE4) in sepsis-related ARDS. One hundred and twenty-three critically ill sepsis patients with/without ARDS and 102 healthy controls were enrolled in this study. Blood samples were collected upon admission for quantitative testing of HE4 by chemiluminescent microparticle immunoassay (CMIA). ROC curve analysis and Spearman's correlation analysis were conducted to determine the diagnostic and prognostic value of HE4. Compared with controls, the serum HE4 concentrations of sepsis patients were elevated, and levels in sepsis patients with ARDS were significantly higher (all p < 0.0001). Moreover, HE4 concentrations were strongly correlated with the clinical severity characteristics of sepsis patients, and ROC curve suggested that the AUC of HE4 applied to discriminate sepsis-ARDS patients from sepsis patients was 0.903. HE4 was also found to be a prognostic biomarker of clinical severity and 28-day mortality among critically ill sepsis patients. Logistic regression analysis showed that HE4 was an independent factor for diagnosis of ARDS. Meanwhile, ROC curve analysis showed that the cut-off value of serum HE4 to discriminate 28-day mortality from sepsis patients (AUC: 0.782) was 646.5pmol/L. The concentration of serum HE4 in patients with sepsis-related ARDS was markedly increased and was significantly correlated with mortality, which suggests that serum HE4 could be a promising diagnostic and prognostic biomarker for ARDS in sepsis patients.

Elevated Serum HE4 Concentrations and Risk of Cardiac Complications among Hospitalized Patients with Burns.

The decrease in effective blood volume after burns is closely related to abnormal heart function. To investigate whether serum human epididymis protein 4 (HE4), an indicator of early renal injury, contributes to increased risk of cardiac complications in patients with burns. Within 24 hours after hospital admission, clinical condition assessment and biochemical testing in patients with burns were performed. Multivariate analysis was performed by evaluating the relationship between serum HE4 levels and risk of cardiac complications (cardiac insufficiency, arrhythmia, and myocardial infarction) during hospitalization. The number (percentage) of cardiac complications in all included patients with burns was 80 (15.6%). The results of sensitivity analysis suggest that elevated serum HE4 levels were related to higher risk of cardiac complications in patients with sepsis (OR = 2.1; 95% CI, 1.19-3.17; P <.001) and in patients without sepsis (OR = 2.29; 95% CI, 1.33-4.71l; P = .005), respectively, after adjustments for clinical confounding factors were made. Sepsis did not have a modification effect on the association between serum and cardiac complications among these patients. Also, the results of ROC curve analysis showed that serum HE4 levels have good predictive value for predicting cardiac complications in patients with burns (AUC = 0.708; 95% CI, 0.61-0.81; P <.001). In the current study, we identified that elevated HE4 levels contributed to increased risk of cardiac complications in the hospital in patients with burns. This novel finding suggests that burn patients with serum HE4 may provide the opportunity to predict cardiac complications before hospital admission.

A clinical study of serum human epididymis protein 4 (HE4) in the diagnosis of pancreatic cancer.

Pancreatic cancer (PC) has poor early diagnosis rates due to its insidious onset. Since human epididymis protein 4 (HE4) is highly expressed in patients with PC, we assessed whether serum HE4 could be a marker for the detection 3 of PC. : Between May 2017 and October 2018, 127 patients with PC were recruited for the study along with 108 healthy controls who underwent health examinations. Serum HE4 concentrations were determined together with levels of carcinoembryonic antigen (CEA) and carbohydrate antigens (CA) 242 (CA242), CA19-9, CA15-3, and CA72-4 by electrochemiluminescence immunoassay (ECLIA) or chemiluminescence immunoassay (CLIA). Correlations between these biomarkers were assessed. : Serum levels of all six biomarkers were higher in patients with PC than in controls (P < 0.05). No correlation was observed between the serum levels of HE4 and the five other tumor markers, although there were strongly significant positive correlations between CA19-9 and CA15-3, and between CA242 and CA72-4. The lack of correlation indicates that HE4 has independent value in the diagnosis of PC. The combined assessment of serum HE4 levels and the other tumor markers improved the sensitivity of diagnosis. In particular, HE4 combined with CA19-9 performed significantly better than HE4 alone, or CA19-9 combined with the other markers. The HE4/CA19-9 combination resulted in 94.49% sensitivity and 99.07% specificity (95% confidence interval: 96.9-100). : HE4 is a biomarker associated with PC with a high specificity , either used alone, or evaluated with other biomarkers together improving the detection of PC. This study may provide a new clinical diagnostic approach for PC detection.

Fluorescence turn-on immunosensing of HE4 biomarker and ovarian cancer cells based on target-triggered metal-enhanced fluorescence of carbon dots

Rapid and sensitive detection of tumor biomarkers and cancer cells is of crucial importance for the early diagnosis and prognosis prediction of cancer. The present report describes a target-induced fluorescence enhancement immunosensor that utilizes the optical property of carbon dots (CDs) and the metal-enhanced fluorescence effect (MEF) property of silver nanoparticles (AgNPs) for the sensitive detection of the cancer biomarker human epididymis protein 4 (HE4) and ovarian cancer cells. Nitrogen and sulfur co-doped CDs with a quantum yield of 85.6% were prepared and served as the fluorophore in MEF. The HE4 antibody (Ab) specific to the HE4 antigen was linked covalently to the surface of the synthesized CDs as the capture. The HE4 Ab-conjugated AgNPs (AgNPs-Ab) were prepared and utilized as signal amplification elements. In the presence of the target HE4, composite sandwich structures were formed between the labeled CDs-Ab and AgNPs-Ab, which brought the CDs and AgNPs into proximity, resulting in the fluorescence of CDs enhancement owing to MEF. The intensity of fluorescence enhancement was positively correlated with the HE4 concentration in the clinically important range of 0.01–200 nM with a limit detection of 2.3 pM. Moreover, the immunosensor was also successfully applied to specific fluorescence labeling and quantitative determination of HE4-positive ovarian cancer cells. The proposed target-triggered MEF sensor platform demonstrated high sensitivity, excellent anti-interference ability, along with successful validation in complex biological matrices, providing a new approach for HE4 detection in early diagnosis and therapeutic monitoring.

Association Between Micronutrient Concentrations and Human Epididymis Protein 4.

BackgroundHuman epididymis protein 4 (HE4) has been frequently used to study in many malignant tumors, while serum nutritional markers are used to determine a person’s health status. However, the link between serum micronutrient concentrations and HE4 has not yet been clarified.MethodsA total of 2464 eligible female participants and serum concentrations of nutritional biomarkers were chosen from the National Health and Nutrition Examination Surveys (NHANES) 2001–2002. For statistical analysis, we used the χ2 test, multivariable linear regression, and analysis of variance. Adjusted models were used, and the concentrations of serum nutritional biomarkers were divided into quartiles.ResultsThe mean age of the participants was 48.07 years. Among twelve micronutrients, five were negatively associated with HE4 in models 1, 2 and 3. Only α-carotene, trans-β-carotene, cis-β-carotene, trans-lycopene and retinol were associated with HE4, with beta coefficients of −0.102, −0.027, −0.506, −0.131 and −0.054, respectively. After performing quartile-based analysis, statistical significance was only found for serum α-carotene, trans-lycopene, and retinol in the three models. In model 3, the beta coefficients [95% confidence intervals (CIs)] of the fourth quartiles compared to the first quartiles for α-carotene, trans-lycopene, and retinol were −3.390 (−5.053, −1.727), −4.036 (−5.722, −2.351) and −4.146 (−5.899, −2.393), respectively. Serum concentrations of these three nutritional biomarkers were inversely related to serum HE4 concentration (p trend <0.001).ConclusionHE4 is a useful and novel biomarker that can be used with many diseases, especially ovarian cancer. Three of our selected micronutrients were inversely associated with HE4 concentration. Supplement of micronutrients may reduce the levels of HE4 and the subsequent of ovarian cancer’s risk. Therefore, a formula that correlates HE4 with nutritional biomarkers needs to be established before use in clinical applications.

Diagnostic Value of Preoperative CA125, LDH and HE4 for Leiomyosarcoma of the Female Reproductive System.

PurposeLeiomyosarcoma (LMS) is a rare and extremely aggressive malignancy that is derived from or shows evidence of differentiation toward smooth muscle. If LMS occurs in the female reproductive system, preoperative diagnosis can be difficult, as LMS is easily mistaken for a uterine leiomyoma, especially a degenerated uterine fibroid (DUF). Thus, we assessed the diagnostic value of the preoperative serum concentrations of cancer antigen 125 (CA125), lactate dehydrogenase (LDH) and human epididymis protein 4 (HE4) for differentiating LMS from DUF.Patients and MethodsWe enrolled patients with LMS or DUF who were receiving treatment in The First Affiliated Hospital of Chongqing Medical University between 2009 and 2020. If the preoperative serum concentrations of CA125, LDH and HE4 of our study participants had been tested, these data were analyzed. The preoperative serum concentrations of CA125, LDH and HE4 in participants with LMS (n = 37) were compared with those of participants with pathologically diagnosed DUF (n = 102), who served as the control group.ResultsThe preoperative serum concentrations of CA125, LDH and HE4 of participants with LMS of the female reproductive system were significantly higher than those of participants with DUF (P = 0.009, P < 0.001, P = 0.001, respectively). The cut-off preoperative serum concentrations of CA125, LDH and HE4 were 30.85 U/mL, 186.50 U/L and 50.50 pmol/L, respectively. When these three parameters were used for an analysis of their combined diagnostic utility, the area under the curve (AUC) was 0.892, the sensitivity was 68.4% and the specificity was 95.1% (P < 0.001).ConclusionA combined analysis of the preoperative serum concentrations of CA125, LDH and HE4 could be a promising method for diagnostically differentiating LMS of the female reproductive system from DUF.

Enhanced Expression of Human Epididymis Protein 4 (HE4) Reflecting Pro-Inflammatory Status Is Regulated by CFTR in Cystic Fibrosis Bronchial Epithelial Cells

Decreased human epididymis protein 4 (HE4) plasma levels were reported in cystic fibrosis (CF) patients under CFTR potentiator ivacaftor therapy, which inversely correlated with lung function improvement. In this study, we investigated whether HE4 expression was affected via modulation of CFTR function in CF bronchial epithelial (CFBE) cells in vitro. HE4 protein levels were measured in the supernatants of CFBE 41o− cells expressing F508del-CFTR or wild-type CFTR (wt-CFTR) after administration of lumacaftor/ivacaftor or tezacaftor/ivacaftor, while HE4 expression in CFBE 41o− cells were also analyzed following application of adenylate cyclase activators Forskolin/IBMX or CFTRinh172. The effect of all of these compounds on CFTR function was monitored by the whole-cell patch-clamp technique. Induced HE4 expression was studied with interleukin-6 (IL-6) in F508del-CFTR CFBE 41o− cells under TNF-α stimulation for 1 h up to 1 week in duration. In parallel, plasma HE4 was determined in CF subjects homozygous for p.Phe508del-CFTR mutation receiving lumacaftor/ivacaftor (Orkambi®) therapy. NF-κB-mediated signaling was observed via the nuclear translocation of p65 subunit by fluorescence microscopy together with the analysis of IL-6 expression by an immunoassay. In addition, HE4 expression was examined after NF-κB pathway inhibitor BAY 11-7082 treatment with or without CFTR modulators. CFTR modulators partially restored the activity of F508del-CFTR and reduced HE4 concentration was found in F508del-CFTR CFBE 41o− cells that was close to what we observed in CFBE 41o− cells with wt-CFTR. These data were in agreement with decreased plasma HE4 concentrations in CF patients treated with Orkambi®. Furthermore, CFTR inhibitor induced elevated HE4 levels, while CFTR activator Forskolin/IBMX downregulated HE4 in the cell cultures and these effects were more pronounced in the presence of CFTR modulators. Higher activation level of baseline and TNF-α stimulated NF-κB pathway was detected in F508del-CFTR vs. wt-CFTR CFBE 41o− cells that was substantially reduced by CFTR modulators based on lower p65 nuclear positivity and IL-6 levels. Finally, HE4 expression was upregulated by TNF-α with elevated IL-6, and both protein levels were suppressed by combined administration of NF-κB pathway inhibitor and CFTR modulators in CFBE 41o− cells. In conclusion, CFTR dysfunction contributes to abnormal HE4 expression via NF-κB in CF.

A homogeneous biosensor for Human Epididymis Protein 4 based on upconversion luminescence resonance energy transfer

Human epididymis protein 4 (HE4) is a widely used biomarker for the early diagnosis of ovarian cancer. Luminescence resonance energy transfer (LRET) is a homogeneous assay technique with great simplicity and specificity, which shows a significant advantage in a bioassay. Lanthanide-doped upconverting nanoparticles (UCNPs) were an ideal material to construct LRET-based probes, because their near-infrared (NIR)-excitable property can effectively eliminate the interference from autoluminescence of biosamples. Herein, a HE4 nanoprobe was fabricated based on LRET strategy, in which UCNPs was employed as an energy donor. The commercial organic dye BHQ-1, whose absorption overlaps well with the blue emission of Tm3+-doped UCNPs, was chosen as an energy acceptor. The two components were tagged with the two ends of a molecular beacon (MB) containing HE4 aptamer sequence. Due to the proximity and spectral overlap of UCNPs and BHQ-1, 82% of upconversion luminescence (UCL) can be quenched through LRET. In presence of HE4, the hairpin structure was opened for the strong interaction between HE4 and its aptamer sequence, which leads to the separation of UCNPs and BHQ-1 and inhibition of LRET. There is a good linear relationship between UCL intensity and the logarithm of HE4 concentration in the range from 0.4 ng/mL to 7.0 ng/mL, with a LOD of 0.021 ng/mL in HEPES buffer. This probe exhibits excellent selectivity and stability, and can also be successfully applied in HE4 quantification in human serum. It might be a reliable method for the early diagnosis of ovarian cancer.

Potential association between elevated serum human epididymis protein 4 and chronic kidney disease in female patients.

Serum levels of human epididymis protein 4 (HE4) are elevated in a large number of women with chronic kidney disease (CKD). However, it remains unclear whether HE4 can be used as a potential biomarker for the diagnosis of CKD. This study aims to determine whether serum HE4 is a potential biomarker for CKD in Han Chinese female patients. A total of 347 Han Chinese female patients aged 19 to 89 were included in the present study. Among these patients, 154 were healthy control individuals, while 193 were hospitalized patients with CKD. Their demographic characteristics were obtained, and the levels of serum creatinine (Scr), beta2-microglobulin (B2M), and cystatin C (CysC) (to assess renal function) were measured. Serum HE4 concentration was determined by electrochemiluminescence. Serum HE4 levels in patients with CKD were significantly higher than those in the control group (P < 0.001). Meanwhile, there were significant differences in HE4 levels among the four CKD subgroups (P < 0.001) via multiple comparisons. In addition, the diagnostic value of HE4 was significantly higher than other indicators by ROC curve analysis. HE4 may not only serve as a potential biomarker to predict CKD but also have an important reference value for CKD staging.

HE4 in Various Body Fluids: A Prospective Pilot Study

Introduction: Human epididymis protein 4 (HE4) is a glycoprotein that is a sensitive and specific serum biomarker for patients with suspected ovarian cancer. HE4 is also secreted in different body fluids such as cervical mucus or urine, which could provide an easy alternative for screening purposes. HE4 protein secretion in different body fluids was analysed in 11 healthy controls and in 10 patients with a benign, 10 with a borderline and 22 with a malignant ovarian tumor. Materials and Methods: Preoperative samples of serum, urine, cervical mucus, saliva, and ascites were collected to measure HE4 with an ELISA assay. Urinary creatinine concentration and cervical total protein concentrations were used as reference analytes, and ratios with HE4 were calculated. Results: Median HE4 concentration in urine was higher in patients with epithelial ovarian cancer (EOC) compared to healthy controls and patients with a benign or borderline mass (p=0.02). HE4/creatinine ratio could differentiate healthy controls from EOC and benign ovarian disease from EOC (AUC 0.76, 95% CI 0.58-0.94 and AUC 0.71, 95% 0.52-0.89, respectively). HE4 in ascites was significantly higher in patients with EOC or borderline ovarian mass compared to patients with a benign ovarian mass (p=0.04). HE4 concentrations in cervical mucus and saliva were not significantly higher in patients with EOC. Conclusion: This study shows that HE4 is abundant in body fluids other than blood and suggests that urinary HE4 levels can be used as a non-invasive diagnostic alternative to detect EOC.

HE4 in the evaluation of tumor load and prognostic stratification of high grade serous ovarian carcinoma

Objective Human epididymis protein 4 (HE4) is a validated, complementary biomarker to cancer antigen 125 (CA125) for high grade serous ovarian carcinoma (HGSC). Currently, there are insufficient data on the utility of longitudinal HE4 measurement during HGSC treatment and follow up. We set to provide a comprehensive analysis on the kinetics and prognostic performance of HE4 with serial measurements during HGSC treatment and follow up. Methods This prospective study included 143 patients with advanced HGSC (ClinicalTrials.gov identifier: NCT01276574). Serum CA125 and HE4 were measured at baseline, before each cycle of chemotherapy and during follow up until first progression. Baseline biomarker values were compared to the tumor load assessed during surgery and to residual disease. Biomarker nadir values and concentrations at progression were correlated to survival. Results The baseline HE4 concentration distinguished patients with a high tumor load from patients with a low tumor load assessed during surgery (p<.0001). The baseline CA125 level was not associated with tumor load to a similar extent (p=.067). At progression, the HE4 level was an independent predictor of worse survival in the multivariate analysis (p=.002). All patients that were alive 3 years post-progression had a serum HE4 concentration below 199.20 pmol/l at the 1st recurrence. Conclusion HE4 is a feasible biomarker in the treatment monitoring and prognostic stratification of patients with HGSC. Specifically, the serum level of HE4 at first relapse was associated with the survival of patients and it may be a useful complementary tool in the selection of second line treatments. This is to the best of our knowledge the first time this finding has been reported.