Cardiac Biomarkers and Iron Status in Hemodialysis Patients

Association of Cardiac Biomarkers of Heart Failure and Iron Status in Hemodialysis Patients

Background:

Peripheral arterial disease (PAD) a condition characterized by atherosclerotic occlusive disease of the lower extremities is commonly observed in patients with chronic kidney disease (CKD) patients, particularly those on dialysis. We conducted detailed biomarkers such as thrombospondin and related inflammatory biomarkers for the risk of developing and presence of PAD. Thrombospondin-4 (TSP-4) is an extracellular matrix protein of the vessel wall. Despite bench evidence, its significance in the clinical setting of chronic kidney disease (CKD) is missing

Methods:

This is a cross-sectional, single-center study. A cohort of 450 patients aged 20 or over, who have been on HD for at least 3 months prior to enrollment (Dec 1, 2021) will be included. TSP-4 and TSP-1 will be measured in HD patients using a commercially available ELISA. PAD is diagnosed by the ankle-brachial index (ABI) We will measure related blood biomarkers such as serum hs-cTnT, N-terminal probrain natriuretic peptide, s-Klotho and FABP-4.

Study Overview

• Status: Completed
• Conditions: Heart Failure; Iron-deficiency

Detailed Description

Study Population and Data Source

This is a cross-sectional, single-center study which will be conducted in the Dialysis Center of Tungs' Taichung MetroHarbor Hospital (TTMHH) in the coastal region of central Taiwan. A cohort of 500 patients aged 20 or over, who have been on HD for at least 3 months prior to enrollment will be included. The medical charts of these patients are reviewed for eligibility identification, and should be compatible with the inclusion/exclusion criteria and enrolled in our analysis. The research was conducted ethically in accordance to the World Medical Association Declaration of Helsinki. Subjects provided written informed consent and the Local Ethical Committee approved the study.

The baseline data such as demographics, comorbidities, anthropometrics, and relevant laboratory data, clinical diagnosis of hear failure based on cardiac echocardiograhy measurements, and medication history will be collected. None of the patients were receiving blood transfusions, or intravenous iron therapy within two weeks before the time of inclusion. Doppler echocardiography using standardized equipment and complying with recommendations from the American Society of Echocardiography (2009)[20]. The biplane method of disks was used to measure left ventricular (LV) EF. Patients were eligible for the study if they had the New York Heart Association (NYHA) functional class II-IV; HFrEF and left ventricular EF (LVEF)≤40%. The characteristics of patients will be exclude from our study were (1) decompensated cirrhosis , (2) neoplastic diseases , (3) incomplete data, (4) receiving hemodialysis < 3 months and active infection.

Iron Status and Other Laboratory Measurements

Blood was drawn with EDTA anticoagulant in the morning after an overnight fast of at least 12 h before a dialysis session. The samples were separated via centrifugation (3000 rpm, 10 min) and immediately stored at -80°C for subsequent assays. Glucose, high-density lipoprotein cholesterol HDL), triglycerides and total cholesterol are all measured by standard assays using the analyzer. Low-density lipoprotein cholesterol (LDL) is measured by standard assay. High-sensitivity C-reactive protein (hsCRP) and are measured using standard nephelometry. Serum hs-cTnT was measured using a sandwich immunoassay method, a novel highly sensitive assay with a lower measurable limit of 3 ng/L.

Iron deficiency was defined using the Kidney Disease Outcomes Quality Initiative guidelines criteria: ferritin < 100 ng/mL or transferrin saturation (TSAT) < 20% when ferritin is < 800 ng/mL.[21] Serum iron was measured using spectrophotometry; serum ferritin and transferrin were measured using immunoturbidimetry. The TSAT was estimated using the formula: TSAT serum iron (μg/dL)/[serum transferrin (mg/dL)×1.25].[22] Additional measures of iron status were: serum soluble transferrin receptor (sTfR) (measured using an enzyme immunoassay),[23-24] and ferritin index. Ferritin index has been proposed as a useful tool in the diagnosis of ID states, where ratios > 2 suggest ID. The ferritin index is calculated by dividing sTfR (expressed in nmol/L or mg/L) by log10 ferritin (measured in ng/mL) and is thus increased whenever sTfR is raised and or ferritin is reduced.[25] Hepcidin is a 25-amino-acid peptide that is produced mainly by the liver, secreted into plasma, and excreted in urine. It is the main regulator of systemic iron homeostasis which restricts the intestinal iron absorption and iron release from macrophages [26]. Serum hepcidin was assayed using a hepcidin ELISA Kit(Human Hepc25), a sandwich-ELISA technique.

Measurement of biomarkers

The GDF15 concentrations are determined by a quantitative sandwich enzyme immunoassay technique (Quantikine®; R&D Systems, Inc. Minneapolis, MN, USA). Quality control (QC) samples from R&D Systems, which had a total coefficient of variation (CV) of 6.3% at low concentrations (159 ng/L), 9.7% at medium concentrations (436 ng/L), and 15.1% at high concentrations (827 ng/L), were included in each assay and results were accepted when QCs fell within manufacturer-specified lot-specific concentration. Overall range of GDF15 detection was 308-13790 ng/L. Gal-3 concentration in plasma was analyzed in duplicate by a commercially available ELISA kit (Norcross, GA or BG Medicine, Inc., Waltham, Mass., USA). Soluble ST2 levels were assessed on baseline samples using a highly sensitive sandwich monoclonal immunoassay (Presage® ST2 Assay, Critical Diagnostics, New York, NY), with a lower limit of detection of 2 ng/mL, an upper limit of detection of 200 ng/mL, an intra-assay coefficient of variation <2.5%, and an interassay coefficient of variation of<4.6%15. Serum H-FABP was measured by an enzyme-linked immunosorbent assay (Hycult Biotech, Uden, the Netherlands). The hsTnT concentrations was measured by electro-chemiluminescence immunoassay using the troponin T high-sensitivity assays, on a Cobas analyser (Roche Diagnostics GmbH, Mannheim, Germany or Abott ). Serum NT-pro-BNP was analyzed using a commercial NT-proBNP ELISA kit. Detection of NT-proBNP ranged from 12 ng/L to 35 000 ng/L and hsTnT ranged between 3 ng/L and 1172 ng/L. All assays were performed in duplicate by technicians blinded to the all clinical data.

• Study Type: Observational
• Enrollment (Actual): 300

Contacts and Locations

Study Locations

• Taiwan
  • Taichung, Taiwan
    • Tungs' Taichung MetroHarbour Hospital

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 20 years and older (Adult, Older Adult)
• Accepts Healthy Volunteers: No
• Genders Eligible for Study: All

• Sampling Method: Probability Sample

Study Population

patients have to be at least 20-years-old and on outpatient hemodialysis (HD) for at least 3 months.

Description

Inclusion Criteria:

1. Both sexes aged 20 years or over
2. Received stable hemodialysis at least 3 months.
3. Written informed consent.

Exclusion Criteria:

1. decompensated cirrhosis.
2. neoplastic diseases.
3. incomplete data.
4. receiving hemodialysis < 3 months and active infection.

Study Plan

How is the study designed?

Design Details

• Observational Models: Case-Crossover
• Time Perspectives: Cross-Sectional

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Measures of iron status	indicating iron storage (ferritin), iron transport (transferrin saturation), regulator of systemic iron homeostasis (hepcidin) and iron demand (soluble transferrin receptor) ,Iron dysmetabolism potentially contributes to heart failure	1 years

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Analysis of biomarkers of GDF-15	The GDF-15 was determined from plasma samples using standardized enzymelinked immunosorbent assay (ELISA) methods	1 years
Analysis of biomarkers of Galectin-3	The Galectin-3 was determined from plasma samples using standardized enzymelinked immunosorbent assay (ELISA) methods	1 years
Analysis of biomarkers of Soluble ST2	The Soluble ST2 was determined from plasma samples using standardized enzymelinked immunosorbent assay (ELISA) methods	1 years
Analysis of biomarkers of hs-cTnT	The hs-cTnT was determined from plasma samples using standardized enzymelinked immunosorbent assay (ELISA) methods	1 years
Analysis of biomarkers of N-terminal probrain natriuretic peptide	The N-terminal probrain natriuretic peptide was determined from plasma samples using standardized enzymelinked immunosorbent assay (ELISA) methods	1 years
Analysis of biomarkers of FABP-4	The FABP-4 was determined from plasma samples using standardized enzymelinked immunosorbent assay (ELISA) methods	1 years

Collaborators and Investigators

• Sponsor: Tungs' Taichung Metroharbour Hospital

Study record dates

Study Major Dates

• Study Start (Actual): June 1, 2022
• Primary Completion (Actual): December 31, 2022
• Study Completion (Actual): December 31, 2022

Study Registration Dates

• First Submitted: February 16, 2022
• First Submitted That Met QC Criteria: February 16, 2022
• First Posted (Actual): February 25, 2022

Study Record Updates

• Last Update Posted (Estimate): February 22, 2023
• Last Update Submitted That Met QC Criteria: February 21, 2023
• Last Verified: February 1, 2023

More Information

Terms related to this study

• Keywords: Heart failure; Iron status; Cardiac biomarkers
• Additional Relevant MeSH Terms: Heart Diseases; Cardiovascular Diseases; Heart Failure
• Other Study ID Numbers: 110070

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No