Exploring the Correlation Between Fibrosis Biomarkers and Clinical Disease Severity in PLN p.Arg14del Patients

Background: Pathogenic variants in phospholamban (PLN, like p. Arg14del), are found in patients diagnosed with arrhythmogenic (ACM) and dilated cardiomyopathy (DCM). Fibrosis formation in the heart is one of the hallmarks in PLN p.Arg14del carriers. During collagen synthesis and breakdown, propeptides are released into the circulation, such as procollagen type I carboxy-terminal propeptide (PICP) and C-terminal telopeptide collagen type I (ICTP).Aim: To investigate if PICP/ICTP levels in blood are correlative biomarkers for clinical disease severity and outcome in PLN p.Arg14del variant carriers.Methods: Serum and EDTA blood samples were collected from 72 PLN p.Arg14del carriers (age 50.5 years, 63% female) diagnosed with ACM (n = 12), DCM (n = 14), and preclinical variant carriers (n = 46). PICP levels were measured with an enzyme-linked immune sorbent assay and ICTP with a radio immuno-assay. Increased PICP/ICTP ratios suggest a higher collagen deposition. Clinical data including electrocardiographic, and imaging results were adjudicated from medical records.Results: No correlation between PICP/ICTP ratios and late gadolinium enhancement (LGE) was found. Moderate correlations were found between the PICP/ICTP ratio and end-diastolic/systolic volume (both rs = 0.40, n = 23, p = 0.06). PICP/ICTP ratio was significantly higher in patients with T wave inversion (TWI), especially in leads V4–V6, II, III, and aVF (p < 0.022) and in patients with premature ventricular contractions (PVCs) during an exercise tolerance test (p = 0.007).Conclusion: High PICP/ICTP ratios correlated with clinical parameters, such as TWI and PVCs. Given the limited size and heterogeneity of the patient group, additional studies are required to substantiate the incremental prognostic value of these fibrosis biomarkers in PLN p.Arg14del patients.

A 3D primary human cell-based in vitro model of non-alcoholic steatohepatitis for efficacy testing of clinical drug candidates

Non-alcoholic steatohepatitis (NASH) is a progressive and severe liver disease, characterized by lipid accumulation, inflammation, and downstream fibrosis. Despite its increasing prevalence, there is no approved treatment yet available for patients. This has been at least partially due to the lack of predictive preclinical models for studying this complex disease. Here, we present a 3D in vitro microtissue model that uses spheroidal, scaffold free co-culture of primary human hepatocytes, Kupffer cells, liver endothelial cells and hepatic stellate cells. Upon exposure to defined and clinically relevant lipotoxic and inflammatory stimuli, these microtissues develop key pathophysiological features of NASH within 10 days, including an increase of intracellular triglyceride content and lipids, and release of pro-inflammatory cytokines. Furthermore, fibrosis was evident through release of procollagen type I, and increased deposition of extracellular collagen fibers. Whole transcriptome analysis revealed changes in the regulation of pathways associated with NASH, such as lipid metabolism, inflammation and collagen processing. Importantly, treatment with anti-NASH drug candidates (Selonsertib and Firsocostat) decreased the measured specific disease parameter, in accordance with clinical observations. These drug treatments also significantly changed the gene expression patterns of the microtissues, thus providing mechanisms of action and revealing therapeutic potential. In summary, this human NASH model represents a promising drug discovery tool for understanding the underlying complex mechanisms in NASH, evaluating efficacy of anti-NASH drug candidates and identifying new approaches for therapeutic interventions.

The Interplay between Myocardial Fibrosis, Strain Imaging and Collagen Biomarkers in Adults with Repaired Tetralogy of Fallot

Background: We sought to assess the interplay between right ventricle (RV) fibrosis, biventricular dysfunction based on global longitudinal strain (GLS) analysis, and biomarkers such as Galectin-3 (Gal-3), procollagen type III (PCIII), and NTproBNP. Methods: We studied 35 adult patients with rToF. All patients underwent a cardiac magnetic resonance (CMR) scan including feature tracking for deformation imaging. Blood biomarkers were measured. Results: LGE RV was detected in all patients, mainly at surgical sites. Patients with the highest RV LGE scoring had greater RV dilatation and dysfunction whereas left ventricular (LV) function was preserved. LV GLS correlated with RV total fibrosis score (p = 0.007). A LV GLS value of −15.9% predicted LGE RV score > 8 (AUC 0.754 (p = 0.02)). Neither RV GLS nor biomarker levels were correlated with the extent of RV fibrosis. A cut-off value for NTproBNP of 145.25 pg/mL predicted LGE RV score > 8 points (AUC 0.729, (p = 0.03)). A cut-off value for Gal-3 of 7.42 ng/mL predicted PR Fraction > 20% [AUC 0.704, (p = 0.05)]. Conclusions: A significant extent of RV fibrosis was mainly detected at surgical sites of RV, affecting RV performance. CMR-FT reveals subtle LV dysfunction in rToF patients, due to decreased performance of the fibrotic RV. Impaired LV function and elevated NTproBNP in rToF reflect a dysfunctional fibrotic RV.

Association between Markers of Fibrosis and Heart Failure Incidence in Patients with Type 2 Diabetes Mellitus

Type 2 diabetes mellitus (T2DM) and chronic heart failure (HF) have close association, and several biomarkers have been studied to better understand this association and improve prediction of HF in T2DM. Furthermore, in recent clinical trials, sodium glucose cotransporter 2 inhibitors (SGLT2i), glucose-lowering drugs, improved HF outcomes. The objective of the present study was to evaluate association between circulating biomarkers of fibrosis and incidence of HF with preserved ejection fraction (HFpEF) in patients with T2DM receiving sodium glucose cotransporter 2 inhibitors (SGLT2i). Materials and Methods. At baseline, transthoracic echocardiography and laboratory assessment of N-terminal fragment of the brain natriuretic peptide (Nt-proBNP), soluble suppression of tumorigenesis-2 (sST2), galectin-3 (Gal-3), C-terminal propeptide of procollagen type I (PICP), N-terminal propeptide of procollagen type III (PIIINP), matrix metalloproteinase-9 (MMP-9), and tissue inhibitor of matrix proteinase-1 (TIMP-1) were done. After 3 years of follow-up, information about HF events (hospitalization for HF, established HF in outpatient department by a cardiologist) was obtained. Results. Seventy-two patients were included in the study. The mean age was 57 (49.7; 63.2) years; 44% were female. Most patients had T2DM for more than 4 years. All patients were overweight or had obesity, and 93% patients had arterial hypertension (AH). After 3 years of follow-up, HFpEF was established in 21% patients. Patients were divided into two groups according to the presence of HFpEF, and baseline characteristics were compared. Patients with HF were older and had longer diabetes and AH duration and higher Nt-proBNP, Gal-3, PIIINP, and PICP levels at baseline than patients without HF (all p < 0.05 ). Gal − 3 > 10 ng/ml ( OR = 2.25 ; 95% CI, 1.88–5.66; p = 0.01 ) and NT − pro − BNP > 80 pg/ml ( OR = 2.64 ; 95% CI, 1.56–4.44; p = 0.001 ) were associated with increased risk of HF incidence. Age > 60 years, diabetes duration > 10 years, and presence of abdominal obesity were independent predictors of HFpEF as well. Conclusions. T2DM patients treated with SLGT2i, who developed HFpEF after 3 years of follow-up, had higher PICP, PIIINP, Gal-3, and NT-proBNP serum concentrations at baseline, and Gal-3 level was an independent predictor of HFpEF.

Serum markers of cardiac fibrosis suffering from heart failure with preserved left ventricular ejection fraction upon ST-segment elevation myocardial infarction

Aim. Currently, there is no method which accurately predicts an adverse outcome of heart failure with a preserved left ventricular ejection fraction (HFpEF) upon ST-segment elevation myocardial infarction (STEMI). Here we studied the prognostic significance of procollagen type I C-terminal propeptide (PICP) and procollagen type III N-terminal propeptide (PIIINP) in patients with post-STEMI HFpEF.Material and Methods. The study included 83 patients (60 men and 23 women) with post-STEMI HFpEF (left ventricular EF ≥ 50%) and 20 ageand gender-matched healthy controls. Serum concentrations of PICP and PIIINP were measured on the 1st day of hospitalization using enzyme-linked immunosorbent assay with the following calculation of PICP/PIIINP ratio.Results. Serum PICP and PIIINP on the 1st day of STEMI significantly (3.4-fold) exceeded the values of the control group and were as follows: PIIINP: 26.0 (18.9; 34.9) ng/mL (р = 0.047); PICP: 609.0 (583.0; 635.0) ng/mL (р = 0.049).Conclusion. Elevated values of procollagens indicate that cardiac fibrosis commences within the 24 hours after STEMI onset. The pivotal role of cardiac fibrosis in the formation of diastolic dysfunction suggests the usefulness of serum procollagens to predict the development of HFpEF in a long-term period upon STEMI.

Profile of Matrix Metalloproteinase Activity, Markers of Collagen and Elastin Degradation and Remodeling During Pregnancy, Delivery, and Puerperium in Pelvic Organ Prolapse 3 Months After Childbirth

Introduction: Pelvic organ prolapse (POP) is a common condition that negatively impacts the quality of life of millions of women. Recent results indicate that a burst of elastic fiber assembly and cross-linking occurs in the vaginal wall postpartum and that synthesis and assembly of elastic fibers are crucial for the recovery of pelvic organ support after vaginal delivery.Methods: A total of 39 primigravida women with gestational age > 36 weeks who underwent vaginal delivery were included in this study. Blood and urine samples were taken for laboratory assessment including carboxyl-terminal telopeptide of the type I collagen (ICTP), procollagen type I N-propeptide (PINP), procollagen type-III N-propeptide (PIIINP), desmosine, tropoelastin, and matrix metalloproteinase-9 (MMP-9). At 24 ˗ 48 hours after labor, the subject was observed in the treatment room and a second blood sample was taken for the examination. At 6 weeks post-partum, a third blood sample was taken for examination.Results: There was a significant relationship between serum MMP-9 levels six weeks post-partum in the pelvic floor dysfunction group and the control group (p = 0.025). In addition, a significant difference was also found in the different levels of MMP-9 (p = 0.041) and tropoelastin (p = 0.041) during delivery and 6 weeks after delivery. There was a significant difference between serum ICTP levels at delivery in the cystocele and control groups (p = 0.042), ICTP (p = 0.019) and tropoelastin (p = 0.046) levels also differ significantly during delivery and 6 weeks after delivery.Conclusion: ICTP, MMP-9, and tropoelastin are potential biomarkers in association with pelvic organ prolapse.

Effects of Six-Week Resistance Training with or without Vibration on Metabolic Markers of Bone Metabolism

Acute and protracted effects of resistive exercise (RE) and resistive exercise with whole-body vibration (RVE) on metabolic markers of bone metabolism were investigated. Twenty-six men participated in a randomized training program including RE (n = 13; age = 23.4 ± 1.4 years) or RVE (n = 13; age = 24.3 ± 3.3 years). During the first session, acute C-terminal telopeptide of type I collagen (CTX) responses decreased by 12.9% (standard deviation, SD 13.7%) after 2 min, followed by a 15.5% (SD 36.0%) increase at 75 min after exercise (both p < 0.001). Procollagen type I amino terminal propeptide (P1NP) increased by 12.9% (SD 9.1%) at 2 min (p < 0.001) but no change occurred at 75 min. Sclerostin showed prolonged responses from 2 to 75 min post-exercise in the first session (p < 0.001). Acute responses at the first session were comparable between groups for CTX and P1NP, acute sclerostin responses were substantially greater in RE than in RVE (p = 0.003). No significant differences were noted in the resting baseline levels of CTX, P1NP, or sclerostin from the beginning to the end of the six-week progressive training. The present study therefore did not demonstrate any sizeable enhancement of bone turnover that could match the effects that have been repeatably made in response to countermeasure exercise during bed rest.

Circulating miR-122-5p and miR-375 as Potential Biomarkers for Bone Mass Recovery after Parathyroidectomy in Patients with Primary Hyperparathyroidism: A Proof-of-Concept Study

Primary hyperparathyroidism (PHPT) is the leading cause of secondary osteoporosis. Although bone mineral density (BMD) tends to recover after parathyroidectomy in PHPT patients, the degree of recovery varies. Circulating microRNAs (miRNAs) profiles are known to be correlated with osteoporosis and fracture. We aimed to investigate whether osteoporotic fracture-related miRNAs are associated with postoperative BMD recovery in PHPT. Here, 16 previously identified osteoporotic fracture-related miRNAs were selected. We analyzed the association between the preoperative level of each miRNA and total hip (TH) BMD change. All 12 patients (among the 18 patients enrolled) were cured of PHPT after parathyroidectomy as parathyroid hormone (PTH) and calcium levels were restored to the normal range. Preoperative miR-19b-3p, miR-122-5p, and miR-375 showed a negative association with the percent changes in TH BMD from baseline. The association remained robust for miR-122-5p and miR-375 even after adjusting for sex, age, PTH, and procollagen type 1 N-terminal propeptide levels in a multivariable model. In conclusion, preoperative circulating miR-122-5p and miR-375 levels were negatively associated with TH BMD changes after parathyroidectomy in PHPT patients. miRNAs have the potential to serve as predictive biomarkers of treatment response in PHPT patients, which merits further investigation.

Relation of Procollagen Type III Amino Terminal Propeptide Level to Sepsis Severity in Pediatrics

Background: Sepsis is still the main etiology of mortality in pediatric intensive care units (PICUs). Therefore, we performed this study to evaluate the value of procollagen Type III amino-terminal propeptide (PIIINP) as a biomarker for sepsis severity diagnosis and mortality. Method: A prospective study was carried out on 170 critically ill children admitted into the PICU and 100 controls. The performed clinical examinations included calculation of the pediatric risk of mortality. Serum PIIINP was withdrawn from patients at admission and from the controls. Results: PIIINP level was significantly more increased in sepsis, severe sepsis, and septic shock than among the controls (p < 0.001). PIIINP was significantly higher in severe sepsis and septic shock (568.3 (32.5–1304.7) and 926.2 (460.6–1370), respectively) versus sepsis (149.5 (29.6–272.9)) (p < 0.001). PIIINP was significantly increased in non-survivors (935.4 (104.6–1370)) compared to survivors (586.5 (29.6–1169)) (p < 0.016). ROC curve analysis exhibited an area under the curve (AUC) of 0.833 for PIIINP, which is predictive for sepsis, while the cut-off point of 103.3 ng/mL had a sensitivity of 88% and specificity of 82%. The prognosis of the AUC curve for PIIINP to predict mortality was 0.651; the cut-off of 490.4 ng/mL had a sensitivity of 87.5% and specificity of 51.6%. Conclusions: PIIINP levels are increased in sepsis, with significantly higher levels in severe sepsis, septic shock, and non-survivors, thus representing a promising biomarker for pediatric sepsis severity and mortality.