Serum-based Biomarkers Research Articles

EP.14A.10 The Impact of Serum-Based Inflammatory Biomarkers on Tumor Proliferation and Prognosis in Malignant Pleural Mesothelioma

EP.14A.10 The Impact of Serum-Based Inflammatory Biomarkers on Tumor Proliferation and Prognosis in Malignant Pleural Mesothelioma

Biomarkers of Hemodynamic Congestion in Heart Failure.

The purpose of this review is to describe the evidence behind various blood and imaging-based biomarkers that can improve the identification of congestion when not clearly evident on routine examination. The natriuretic peptides (NPs) BNP and NT-proBNP have been shown to closely correlate with intra-cardiac filling pressures, both at baseline and when trended following improvement in congestion. Additionally, NPs rise well before clinical congestion is apparent so can be used as a tool to help identify subclinical HF decompensation. Additional serum-based biomarkers including MR-proANP and CA-125 can be helpful in assisting with diagnostic certainty when BNP or NT-proBNP are in the "grey zone" or when factors are present which may confound NP levels. Additionally, the emerging use of ultrasound techniques may enhance our ability to fine-tune the assessment and treatment of congestion. Biomarkers, including the blood-based natriuretic peptides and markers on bedside point of care ultrasound, can be used as non-invasive indices of hemodynamic congestion. These biomarkers are particularly valuable to incorporate when the degree of a patient's congestion is not apparent on clinical exam, and they can provide important prognostic information and help guide clinical management.

Development, External Validation, and Biomolecular Corroboration of Interoperable Models for Identifying Critically Ill Children at Risk of Neurologic Morbidity.

Declining mortality in the field of pediatric critical care medicine has shifted practicing clinicians' attention to preserving patients' neurodevelopmental potential as a main objective. Earlier identification of critically ill children at risk for incurring neurologic morbidity would facilitate heightened surveillance that could lead to timelier clinical detection, earlier interventions, and preserved neurodevelopmental trajectory. Develop machine-learning models for identifying acquired neurologic morbidity while hospitalized with critical illness and assess correlation with contemporary serum-based, brain injury-derived biomarkers. Retrospective cohort study. Two large, quaternary children's hospitals. Critical illness. The outcome was neurologic morbidity, defined according to a computable, composite definition at the development site or an order for neurocritical care consultation at the validation site. Models were developed using varying time windows for temporal feature engineering and varying censored time horizons prior to identified neurologic morbidity. Optimal models were selected based on F1 scores, cohort sizes, calibration, and data availability for eventual deployment. A generalizable created at the development site was assessed at an external validation site and optimized with spline recalibration. Correlation was assessed between development site model predictions and measurements of brain biomarkers from a convenience cohort. After exclusions there were 14,222-25,171 encounters from 2010-2022 in the development site cohorts and 6,280-6,373 from 2018-2021 in the validation site cohort. At the development site, an extreme gradient boosted model (XGBoost) with a 12-hour time horizon and 48-hour feature engineering window had an F1-score of 0.54, area under the receiver operating characteristics curve (AUROC) of 0.82, and a number needed to alert (NNA) of 2. A generalizable XGBoost model with a 24-hour time horizon and 48-hour feature engineering window demonstrated an F1-score of 0.37, AUROC of 0.81, AUPRC of 0.51, and NNA of 4 at the validation site. After recalibration at the validation site, the Brier score was 0.04. Serum levels of the brain injury biomarker glial fibrillary acidic protein measurements significantly correlated with model output (r s =0.34; P =0.007). We demonstrate a well-performing ensemble of models for predicting neurologic morbidity in children with biomolecular corroboration. Prospective assessment and refinement of biomarker-coupled risk models in pediatric critical illness is warranted. Question Can interoperable models for predicting neurological deterioration in critically ill children be developed, correlated with serum-based brain-derived biomarkers, and validated at an external site? Findings A development site model demonstrated an area under the receiver operating characteristics curve (AUROC) of 0.82 and a number needed to alert (NNA) of 2. Predictions correlated with levels of glial fibrillary acidic protein in a subset of children. A generalizable model demonstrated an AUROC of 0.81 and NNA of 4 at the validation site. Meaning Well performing prediction models coupled with brain biomarkers may help to identify critically ill children at risk for acquired neurological morbidity.

Discovery of crucial cytokines associated with deep vein thrombus formation by protein array analysis

BackgroundExpanding the number of biomarkers is imperative for studying the etiology and improving venous thromboembolism prediction. In this study, we aimed to identify promising biomarkers or targeted therapies to improve the detection accuracy of early-stage deep vein thrombosis (DVT) or reduce complications.MethodsQuantibody Human Cytokine Antibody Array 440 (QAH-CAA-440) was used to screen novel serum-based biomarkers for DVT/non-lower extremity DVT (NDVT). Differentially expressed proteins in DVT were analyzed using bioinformatics methods and validated using a customized array. Diagnostic accuracy was calculated using receiver operating characteristics, and machine learning was applied to establish a biomarker model for evaluating the identified targets. Twelve targets were selected for validation.ResultsCytokine profiling was conducted using a QAH-CAA-440 (RayBiotech, USA) quantimeter array. Cross-tabulation analysis with Venn diagrams identified common differential factors, leading to the selection of 12 cytokines for validation based on their clinical significance. These 12 biomarkers were consistent with the results of previous array analysis: FGF-6 (AUC = 0.956), Galectin-3 (AUC = 0.942), EDA-A2 (AUC = 0.933), CHI3L1 (AUC = 0.911), IL-1 F9 (AUC = 0.898), Dkk-4 (AUC = 0.88), IG-H3 (AUC = 0.876), IGFBP (AUC = 0.858), Gas-1 (AUC = 0.858), Layilin (AUC = 0.849), ULBP-2 (AUC = 0.813)and FGF-9 (AUC = 0.773). These cytokines are expected to serve as biomarkers, targets, or therapeutic targets to differentiate DVT from NDVT.ConclusionsEDA-A2, FGF-6, Dkk-4, IL-1 F9, Galentin-3, Layilin, Big-h3, CHI3L1, ULBP-2, Gas-1, IGFBP-5, and FGF-9 are promising targets for DVT diagnosis and treatment.

Alterations of human CSF and serum-based mitophagy biomarkers in the continuum of Alzheimer disease

ABSTRACT Defective mitophagy is consistently found in postmortem brain and iPSC-derived neurons from Alzheimer disease (AD) patients. However, there is a lack of extensive examination of mitophagy status in serum or cerebrospinal fluid (CSF), and the clinical potential of mitophagy biomarkers has not been tested. We quantified biomarkers of mitophagy/autophagy and lysosomal degradation (PINK1, BNIP3L and TFEB) in CSF and serum from 246 individuals, covering mild cognitive impairment due to AD (MCI-AD, n = 100), dementia due to AD (AD-dementia, n = 100), and cognitively unimpaired individuals (CU, n = 46), recruited from the Czech Brain Aging Study. Cognitive function and brain atrophy were also assessed. Our data show that serum and CSF PINK1 and serum BNIP3L were higher, and serum TFEB was lower in individuals with AD than in corresponding CU individuals. Additionally, the magnitude of mitophagy impairment correlated with the severity of clinical indicators in AD patients. Specifically, levels of PINK1 positively correlated with phosphorylated (p)-MAPT/tau (181), total (t)-MAPT/tau, NEFL (neurofilament light chain), and NRGN (neurogranin) levels in CSF and negatively with memory, executive function, and language domain. Serum TFEB levels negatively correlated with NEFL and positively with executive function and language. This study reveals mitophagy impairment reflected in biofluid biomarkers of individuals with AD and associated with more advanced AD pathology. Abbreviation: Aβ: amyloid beta; AD: Alzheimer disease; AVs: autophagic vacuoles; BNIP3L: BCL2 interacting protein 3 like; CU: cognitively unimpaired; CSF: cerebrospinal fluid; LAMP1: lysosomal-associated membrane protein 1; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MCI: mild cognitive impairment; NRGN: neurogranin; NEFL: neurofilament light chain; p-MAPT/tau: phosphorylated microtubule associated protein tau; PINK1: PTEN induced kinase 1; t-MAPT/tau: total microtubule associated protein tau; TFEB: transcription factor EB; TMT: Trail Making Test.

Association between serum creatinine to albumin ratio and short- and long-term all-cause mortality in patients with acute pancreatitis admitted to the intensive care unit: a retrospective analysis based on the MIMIC-IV database.

Serum creatinine (Cr) and albumin (Alb) are important predictors of mortality in individuals with various diseases, including acute pancreatitis (AP). However, most previous studies have only examined the relationship between single Cr or Alb levels and the prognosis of patients with AP. To our knowledge, the association between short- and long-term all-cause mortality in patients with AP and the blood creatinine to albumin ratio (CAR) has not been investigated. Therefore, this study aimed to evaluate the short- and long-term relationships between CAR and all-cause mortality in patients with AP. We conducted a retrospective study utilizing data from the Medical Information Market for Intensive Care (MIMIC-IV) database. The study involved analyzing various mortality variables and obtaining CAR values at the time of admission. The X-tile software was used to determine the optimal threshold for the CAR. Kaplan-Meier (K-M) survival curves and multivariate Cox proportional hazards regression models were used to assess the relationship between CAR and both short- and long-term all-cause mortality. The predictive power, sensitivity, specificity, and area under the curve (AUC) of CAR for short- and long-term mortality in patients with AP after hospital admission were investigated using Receiver Operating Characteristic analysis. Additionally, subgroup analyses were conducted. A total of 520 participants were included in this study. The CAR ideal threshold, determined by X-tile software, was 0.446. The Cox proportional hazards model revealed an independent association between CAR≥0.446 and all-cause mortality at 7-day (d), 14-d, 21-d, 28-d, 90-d, and 1-year (y) before and after adjustment for confounders. K-M survival curves showed that patients with CAR≥0.446 had lower survival rates at 7-d, 14-d, 21-d, 28-d, 90-d, and 1-y. Additionally, CAR demonstrated superior performance, with higher AUC values than Cr, Alb, serum total calcium, Glasgow Coma Scale, Systemic Inflammatory Response Syndrome score, and Sepsis-related Organ Failure Assessment score at 7-d, 14-d, 21-d, 28-d, 90-d, and 1-y intervals. Subgroup analyses showed that CAR did not interact with a majority of subgroups. The CAR can serve as an independent predictor for short- and long-term all-cause mortality in patients with AP. This study enhances our understanding of the association between serum-based biomarkers and the prognosis of patients with AP.

Cancer-specific risk prediction with a serum microRNA signature.

We recently derived and validated a serum-based microRNA risk score (miR-score) that predicted colorectal cancer (CRC) occurrence with very high accuracy within 14 years of follow-up in a population-based cohort study from Germany (ESTHER cohort). Here, we aimed to evaluate associations of the CRC-specific miR-score with the risk of developing other common cancers, including female breast cancer (BC), lung cancer (LC), and prostate cancer (PC), in the ESTHER cohort. MicroRNAs (miRNAs) were profiled by quantitative real-time PCR in serum samples collected at baseline from randomly selected incident cases of BC (n = 90), LC (n = 88), and PC (n = 93) and participants without diagnosis of CRC, LC, BC, or PC (controls, n = 181) until the end of the 17-year follow-up. Multivariate logistic regression models were used to evaluate the associations of the miR-score with BC, LC, and PC incidence. The miR-score showed strong inverse associations with BC and LC incidence [odds ratio per 1 standard deviation increase: 0.60 (95% confidence interval [CI] 0.43-0.82), p = 0.0017, and 0.64 (95% CI 0.48-0.84),p = 0.0015, respectively]. Associations with PC were not statistically significant but pointed in the positive direction. Our study highlights the potential of serum-based miRNA biomarkers for cancer-specific risk prediction. Further large cohort studies aiming to investigate, validate, and optimize the use of circulating miRNA signatures for cancer risk assessment are warranted.

Serum-based biomarkers associated with lung cancer risk and cause-specific mortality in the German randomized Lung Cancer Screening Intervention (LUSI) trial.

Lung cancer (LC) screening can be optimized using individuals' estimated risks of having a detectable lung tumor, as well as of mortality risk by competing causes, to guide decisions on screening eligibility, ideal screening intervals and stopping ages. Besides age, sex and smoking history, blood-based biomarkers may be used to improve the assessment of LC risk and risk of mortality by competing causes. In the German randomized Lung Screening Intervention Trial (LUSI), we measured growth/differentiation factor-15 (GDF-15), interleukin-6 (IL-6), C-reactive protein (CRP) and N-terminal pro-brain natriuretic protein (NT-proBNP), in blood serum samples collected at start of the trial. Participants in the computed tomography (CT)-screening arm also had a pulmonary function test. Regression models were used to examine these markers as predictors for impaired lung function, LC risk and mortality due to LC or other causes, independently of age, sex and smoking history. Our models showed increases in LC risk among participants with elevated serum levels of GDF-15 [odds ratio (OR)Q4-Q1 =2.47, 95% confidence interval (CI): 1.49-4.26], IL-6 [ORQ4-Q1 =2.36 (1.43-4.00)] and CRP [ORQ4-Q1 =1.81 (1.08-2.75)]. Likewise, proportional hazards models showed increased risks for LC-related mortality, hazard ratio (HR)Q4-Q1 of 4.63 (95% CI: 2.13-10.07) for GDF-15, 3.56 (1.72-7.37) for IL-6 and 2.34 (1.24-4.39) for CRP. All four markers were associated with increased risk of mortality by causes other than LC, with strongest associations for GDF-15 [HRQ4-Q1 =3.04 (2.09-4.43)] and IL-6 [HRQ4-Q1 =2.98 (2.08-4.28)]. Significant associations were also observed between IL-6, CRP, GDF-15 and impaired pulmonary function [chronic obstructive pulmonary disease (COPD), preserved ratio impaired spirometry (PRISm)]. Multi-marker models identified GDF-15 and IL-6 as joint risk predictors for risk of LC diagnosis, without further discrimination by CRP or NT-proBNP. A model based on age, sex, smoking-related variables, GFD-15 and IL-6 provided moderately strong discrimination for prediction of LC diagnoses within 9 years after blood sampling [area under the curve (AUC) =74.3% (57.3-90.2%)], compared to 67.0% (49.3-84.8%) for a model without biomarkers. For mortality by competing causes, a model including biomarkers resulted in an AUC of 76.2% (66.6-85.3%)], compared to 70.0% (60.9-77.9%) a model including age, sex and smoking variables. Serum GDF-15 and IL-6 may be useful indicators for estimating risks for LC and competing mortality among long-term smokers participating in LC screening, to optimize LC screening strategies.

The Relation Between Serum-based Systemic Inflammatory Biomarkers and Locoregional Lymph Node Metastasis in Clinical Stages I to II Right-sided Colon Cancers: The Role of Platelet-to-Lymphocyte Ratio.

It aimed to evaluate the relationship between the systemic inflammatory markers and the lymph node metastasis in clinical stages I to II right-sided colon cancers. A total of 51 eligible clinical stages I to II right-side located colon cancer patients were included in the study. Complete mesocolic excision and central vascular ligation procedures were performed in all cases. All the patients were divided into 2 main groups, pN - (n = 22) and pN + (n = 29). Demographic parameters, preoperative serum-based inflammatory biomarkers, and histopathological findings were compared between the groups. The mean age was 61.0 (54 to 71) years. Of the patients, 51.0% (26/51) were females. The open surgical approach was performed on 54.9% (28/51) of the patients and 45.1% (23/51) was performed laparoscopy. The mean total number of retrieved lymph nodes was 29.1. The lympho-vascular invasion was significantly higher in the pN + group (89.7% vs 50.0%). There were no significant differences in neutrophil-to-lymphocyte ratio, C-reactive protein-to-albumin ratio, mean platelet volume-to-platelet ratio, hemoglobine-albuminelymphocyte-platelet score, systemic inflammation index, lymphocyte-to-monocyte ratio, neutrophil-to-monocyte ratio, lymphocyte-to-C-reactive protein ratio (LCR), neutrophil-to-albumin ratio, and prognostic nutritional index. However, the mean platelet-to-lymphocyte ratio (PLR) was significantly lower in the pN + group (pN - : 282.1 vs pN + : 218.7, P = 0.048). The cutoff value for PLR was determined as 220 according to receiver operating characteristic analysis, with a 63.6% sensitivity and 65.6% specificity. Although it has limited sensitivity and specificity, decreased preoperative PLR was significantly associated with lymph node metastasis in patients with clinical stages I to II right-sided colon cancer. It should be considered as a biomarker for nodal involvement when planning treatment strategies.

Lorpucitinib (JNJ-64251330) in patients with familial adenomatous polyposis (FAP): Results from a phase 1b study.

10514 Background: FAP is the most common hereditary adenomatous polyposis syndrome and, if left untreated, will lead to colorectal cancer development in nearly all patients. Chronic inflammation, driven in part by the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway, may promote polyp formation. Enteric-selective inhibition of the JAK/STAT pathway may provide a treatment for FAP. Lorpucitinib is an oral, small molecule, potent pan-JAK inhibitor with favorable enteric-selective properties. Here, we report the efficacy, safety, and pharmacokinetic (PK) results from a phase 1b study of lorpucitinib in patients with FAP (NCT05014360). Methods: Patients aged ≥18 years with ≥6 polyps ≥2 mm in diameter in the rectum or colon with FAP ( APC germline mutation or obligate carrier) received lorpucitinib 75 mg twice daily for 24 weeks. The primary objective was to evaluate the effect of lorpucitinib on colorectal polyp burden at Week 24, assessed by endoscopy at baseline and Week 24. Secondary objectives were safety, PK, and biomarker assessments of lorpucitinib activity. Results: Overall, 42 (23 pre-colectomy; 19 post-colectomy) patients with FAP were enrolled, with a median age of 33 (range, 19-61) years. No clear effect on polyp burden was observed at Week 24. Gut mucosal and polyp concentrations of lorpucitinib were much higher than plasma concentrations, indicating an enteric-selective distribution. Mean sigmoid colon, rectum, and polyp concentrations of lorpucitinib at Week 8 were 112-, 473-, and 108-fold higher, respectively, than the observed maximum plasma concentration at Week 8 (1 hour). At Week 8, lorpucitinib achieved JAK tyrosine kinase inhibition in mucosal biopsies as evidenced by a median 37% reduction in normalized pSTAT-3 levels. Lorpucitinib reduced serum-based inflammatory biomarkers such as C-reactive protein and serum amyloid A by Week 16 of therapy. Some patients reported an improvement of clinical symptoms such as less frequent bowel movements and less blood in the stool. The most common adverse events (AEs) were abdominal pain (16%), bilirubin increased (13%), headache (13%), lipase increased (13%), and diarrhea (13%). There were no treatment-related serious AEs, grade 3 treatment-emergent AEs, serious infections, or venous thromboembolisms. Conclusions: These results show that lorpucitinib had no clear effect on polyp burden. The study found that lorpucitinib is safe and exhibits an enteric-selective distribution in FAP patients. Inhibition of JAK tyrosine kinase signaling was associated with reduced STAT-3 phosphorylation in mucosal biopsies from FAP patients. While reductions observed in polyp burden were not clinically significant, lorpucitinib treatment lowered levels of several serum inflammatory biomarkers. Treatment with lorpucitinib was safe and well tolerated, and may improve clinical symptoms associated with FAP. Clinical trial information: NCT05014360 .

Performance of serum-based biomarkers in fibrosis prediction of chronic hepatitis B patients in real life practice.

Few studies have addressed the performance and diagnostic accuracy of laboratory-based markers for fibrosis prediction in chronic hepatitis B (CHB) patients yielding heterogeneous results. We aimed to study the performance of the FIB-4 and neutrophil-to-lymphocyte ratio (NLR) markers for the differentiation between significant and non-significant hepatic fibrosis in real-life practice. We prospectively recruited CHB patients attending the hepatology clinic to undergo shear wave elastography (SWE) and blood tests. The predictive accuracy of FIB-4 and NLR for liver fibrosis was assessed by receiver operating characteristics (ROC) analysis. Overall, 174 fully characterized CHB patients with an average age of 50.2±11.2 (29-86 years) and a male predominance (65.2%) were included. Of these, 23% had significant fibrosis (≥F2) per SWE (>7.1KPA). A significant and linear correlation was found between the SWE score and FIB-4 values (r=0.572; P<0.001). A lower cut-off of 1.43 has yielded an AUROC of 0.76, with a sensitivity of 68.8%, specificity of 79.8%, diagnostic accuracy of 78.5%, and NPV of 96%. On the contrary, NLR values were similar between significant and minimal fibrosis and were not found to be correlated with significant fibrosis (r=0.54, P=0.39). FIB4 has a moderate performance and may have a valuable role in excluding significant fibrosis in CHB patients in daily practice.

A multiplex DNA methylation assay for noninvasive early detection of gastric cancer.

300 Background: Gastric cancer (GC) is the fifth leading cancer that ranked fourth in cancer mortality worldwide. Early screening and detection can significantly improve patients’ prognosis and survival. Although in standard GC diagnosis, gastroscopy provides sufficient diagnostic power for identifying GC in early stages, the invasive and costly procedure is not suitable for screening and has low compliance in countries with high GC incidence and large populations. Furthermore, the serum-based GC related biomarkers and non-invasive tests (serum pepsinogen I and II, gastrin-17, and anti-Helicobacter pylori IgG antibody tests) showed insufficient evidence as cost-effective GC screening approaches. A non-invasive DNA methylation assay with high sensitivity was aimed to develop and validate as a potential alternative for GC early detection. Methods: A multiplex quantitative PCR (qPCR) assay for detecting GC-specific DNA methylation markers in plasma was developed, in which the markers were further optimized from a reported seven-marker panel based on a pilot case-control study with 63 GC and 130 non-GC samples. The GC detection model of the assay was developed and locked down in the first cohort consisting of 54 GC and 79 non-GC samples. The second validation cohort with 117 GC and 309 non-GC samples was used to evaluate the assay performance. All plasma samples used in the two cohorts were prospectively collected from patients diagnosed with GC or other gastrointestinal diseases or from healthy donors. Results: The multiplex DNA methylation assay showed an overall sensitivity and specificity of 80% (95% CI: 77–83%) and 65% (95% CI: 63–66%) in the model development cohort, respectively, with a sensitivity of 80% (95% CI: 65–95%, n=10) in the detection of stage I GC. The assay revealed similar performance characteristics in the validation cohort, with an overall sensitivity of 82% (95% CI: 80–83%), a specificity of 69% (95% CI: 68–69%), and a sensitivity of 78% (95% CI: 74–82%, n=41) for the stage I GC detection. The DNA methylation assay outperformed the reported serum-based GC related biomarkers and non-invasive tests (sensitivity of 69.6-70.7% and specificity of 66.8-67.8%) with higher sensitivity, particularly in the stage I GC with a comparable specificity. Conclusions: The multiplex DNA methylation assay provides a potential cost-effective tool that features high sensitivity for GC early detection.

LncRNA T376626 is a promising serum biomarker and promotes proliferation, migration, and invasion via binding to LAMC2 in triple-negative breast cancer

PurposeCirculating long noncoding RNAs (lncRNAs) have been reported to serve as biomarkers for cancer diagnosis. Here, we identified the clinical diagnostic value and biological function of lncRNA T376626 in triple-negative breast cancer (TNBC). MethodA genome-wide lncRNA microarray was used to screen promising serum-based lncRNA biomarkers. The expression of candidate serum lncRNAs was validated in 282 breast cancer (BC) patients and 78 healthy subjects. The diagnostic value of serum lncRNA T376626 was determined by receiver operating characteristic (ROC) curve. RNA fluorescent in situ hybridization (FISH) and RNAScope ISH assays were conducted to examine the expression and localization of lncRNA T376626 in TNBC cells and BC tissues. Kaplan-Meier analysis was conducted to evaluate the relationship between lncRNA T376626 and BC patients’ overall survival (OS) rate. CCK-8, colony-forming, wound healing and Transwell assays were performed to investigate the biological function of lncRNA T376626 on cell proliferation, migration, and invasion in two TNBC cell lines. Cell apoptosis-, cell cycle- and epithelial-mesenchymal transition (EMT)-related biomarkers were quantified by western blots. The lncRNA T376626 binding proteins were screened and identified by RNA pulldown. ResultsLncRNA T376626 level was significantly higher in TNBC serums and tissues. Higher levels of lncRNA T376626 were positively associated with a higher pathological differentiation stage, more aggressive molecular subtype, and poor prognosis in BC and TNBC patients. The area under the curve (AUC) of serum lncRNA T376626 was 0.842. Overexpression (Knockdown) of lncRNA T376626 significantly promoted (inhibited) TNBC cell proliferation, migration, and invasion, possibly by regulating several cell cycle, cell apoptosis and EMT biomarkers. LAMC2 were identified as lncRNA T376626-binding proteins. LAMC2 facilitated TNBC proliferation and metastasis through lncRNA T376626. ConclusionsLncRNA T376626 may serve as a TNBC serum-based diagnostic and prognostic biomarker and play an oncogenic role in TNBC progression through binding to LAMC2.

Improved colorectal cancer screening by adding noninvasive serum-based biomarkers

Background Colorectal cancer (CRC) is one of the most common causes of cancer-related morbidity and mortality worldwide. The most widely used CRC diagnostic tests either noninvasive or invasive are limited by insufficient performance or patient compliance. Profiling inflammatory mediators with carcinogenic roles could aid in CRC diagnosis. We aimed to use multiplex bead assay in evaluating the utility of serum eotaxin-1, macrophage-inflammatory protein-1β (MIP-1β), granulocyte colony-stimulating factor (G-CSF), vascular endothelial growth factor A (VEGF-A), and Fas ligand (FasL) as potential biomarkers in CRC. Patients and methods The study was conducted on 87 patients. Based on colonoscopy findings, patients were divided into 35 CRC and 52 nonmalignant patients (nine with colon polyp, 24 with colitis, and 19 with normal mucosa). Multiplex assay was used for measuring the studied biomarkers. Results The median values of eotaxin-1, MIP-1β, G-CSF, and VEGF-A were significantly higher in patients with CRC compared with the nonmalignant group. The area under the receiver operating characteristics curve for eotaxin-1, MIP-1β, G-CSF, and VEGF-A was 0.863. The area under the ROC for occult blood in stool was only 0.597. Moreover, significantly higher levels of G-CSF and VEGF-A were found in patients with CRC than the precancerous colon polyp group. Conclusions Serum profiling of eotaxin-1, MIP-1β, G-CSF, and VEGF-A could be used as potential biomarkers in early CRC diagnosis with better discriminatory power than stool occult blood and may increase occult blood performance, thus reducing false-positives rates and unneeded colonoscopy. Multiplexing bead technology represents a promising approach for CRC screening and diagnosis.

Examining the Differences in Biomarkers of Neuronal and Glial Injury Between Autoimmune Neurologic Disease Patients and Healthy Controls

Objective To evaluate differences in concentrations of serum-based biomarkers obtained from a screened healthy control (HC) population compared to age and sex matched autoimmune and inflammatory neurologic disease (AIND) patients. Background Protein markers of neuronal and glial injury have become important, minimally invasive biomarkers for understanding the impact of disease in various neurological disorders, including ongoing research into AINDs. Levels of these proteins in healthy individuals remain unclear. Design/Methods Neurofilament light (NfL), glial fibrillary acidic protein (GFAP), ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1), and total tau (Tau) levels in AIND and HC participants will be compared. Multiple Sclerosis (MS) patients, as inflammatory controls, will be analyzed as well. AIND and HC participant serum samples have been collected to obtain NfL, GFAP, UCH-L1, and Tau protein levels using Quanterix SR-XTM SIMOA. We plan to analyze and compare age and sex matched HC samples to AIND patients for each biomarker, ages ranging from 20-80 years old. Concentrations will be log transformed and analyzed with mixed model regression. Results Healthy Controls (130), AIND (255), and newly diagnosed/treatment naïve MS samples (681) have already been collected. Nineteen HC and AIND (includes NMDA, LGI1, TRIM46, CRMP5, MOG, DPPX, GABA-A, GAD-65) participants were analyzed. Preliminary results for HC show mean Nfl (10.3 pg/mL), GFAP (78.9 pg/mL), UCH-L1 (5.65 pg/mL), and Tau (0.53 pg/mL) levels, and for AIND patients mean Nfl (186.48 pg/mL), GFAP (434.92 pg/mL), UCH-L1 (71.38 pg/mL), and Tau (51.85 pg/mL) levels. While ongoing biomarker analysis will be completed with HC that are age and sex matched, the significantly higher levels in AIND patients highlights the importance of creating baseline values in HC to understand these same biomarkers in AIND patients. Conclusions Preliminary results show NfL and GFAP levels are significantly higher in AIND patients versus HC. Baseline biomarker values are essential for understanding further research in biomarkers related to AIND.

Identification of potential microRNA diagnostic panels and uncovering regulatory mechanisms in breast cancer pathogenesis

Early diagnosis of breast cancer (BC), as the most common cancer among women, increases the survival rate and effectiveness of treatment. MicroRNAs (miRNAs) control various cell behaviors, and their dysregulation is widely involved in pathophysiological processes such as BC development and progress. In this study, we aimed to identify potential miRNA biomarkers for early diagnosis of BC. We also proposed a consensus-based strategy to analyze the miRNA expression data to gain a deeper insight into the regulatory roles of miRNAs in BC initiation. Two microarray datasets (GSE106817 and GSE113486) were analyzed to explore the differentially expressed miRNAs (DEMs) in serum of BC patients and healthy controls. Utilizing multiple bioinformatics tools, six serum-based miRNA biomarkers (miR-92a-3p, miR-23b-3p, miR-191-5p, miR-141-3p, miR-590-5p and miR-190a-5p) were identified for BC diagnosis. We applied our consensus and integration approach to construct a comprehensive BC-specific miRNA-TF co-regulatory network. Using different combination of these miRNA biomarkers, two novel diagnostic models, consisting of miR-92a-3p, miR-23b-3p, miR-191-5p (model 1) and miR-92a-3p, miR-23b-3p, miR-141-3p, and miR-590-5p (model 2), were obtained from bioinformatics analysis. Validation analysis was carried out for the considered models on two microarray datasets (GSE73002 and GSE41922). The model based on similar network topology features, comprising miR-92a-3p, miR-23b-3p and miR-191-5p was the most promising model in the diagnosis of BC patients from healthy controls with 0.89 sensitivity, 0.96 specificity and area under the curve (AUC) of 0.98. These findings elucidate the regulatory mechanisms underlying BC and represent novel biomarkers for early BC diagnosis.

Expression analysis of circulating miR-22, miR-122, miR-217 and miR-367 as promising biomarkers of acute lymphoblastic leukemia.

The role of serum-based biomarkers such as microRNAs in cancer diagnosis has been extensively established. This study aimed to determine the expression levels of bioinformatically selected miRNAs and whether they can be used as biomarkers or a new therapeutic target in patients with acute lymphoblastic leukemia (ALL). The expression levels of serum miR-22, miR-122, miR-217, and miR-367 in 21 ALL patients and 21 healthy controls were measured using quantitative real-time PCR. The receiver operating characteristic (ROC) curve and the associated area under the curve (AUC) was used to assess candidate miRNAs' diagnostic value as a biomarker. The results showed that miR-217 was markedly decreased in patients with ALL compared to controls. Moreover, miR-22, miR-122, and miR-367 were found to be upregulated. Furthermore, ROC analysis showed that serum miR-217 and miR-367 could differentiate ALL patients from healthy individuals, while miR-22 has approximate discriminatory power that requires further investigation. These results provide promising preliminary evidence that circulating miR-217 and miR-367 could be considered potent diagnostic biomarkers and therapeutic goals in this disease.

Biomarkers in Liquid Biopsies for Prediction of Early Liver Metastases in Pancreatic Cancer.

Simple SummaryIndividualized diagnostics approaches in modern cancer therapy require predictive and prognostic biomarkers that are easily accessible and stratify patients for optimal and individualized treatment. Pancreatic ductal adenocarcinoma (PDAC) is still a life-threatening disease mainly because of its late diagnosis in advanced stages or rapid progress even in patients with curative resection of the primary tumor. Moreover, patients with liver metastases exhibit an even worse prognosis. Hence, this retrospective multi-center study aims to identify biomarkers in perioperative serum of PDAC patients predicting early liver metastasis. A highly sensitive biomarker analysis was performed using two different methodological approaches. Olink® analysis, which was also used to validate LEGENDplexTM results, identified significant differences in proteins involved in chemotaxis and migration of immune cells as well as cell growth in serum of patients with early versus late onset of liver metastasis. Further studies with larger cohorts are required to validate these findings for clinical translation.Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive solid malignancies with poor survival rates. Only 20% of the patients are eligible for R0-surgical resection, presenting with early relapses, mainly in the liver. PDAC patients with hepatic metastases have a worse outcome compared to patients with metastases at other sites. Early detection of hepatic spread bears the potential to improve patient outcomes. Thus, this study sought for serum-based perioperative biomarkers allowing discrimination of early (EHMS ≤ 12 months) and late hepatic metastatic spread (LHMS > 12 months). Serum samples from 83 resectable PDAC patients were divided into EHMS and LHMS and analyzed for levels of inflammatory mediators by LEGENDplexTM, which was validated and extended by Olink® analysis. CA19-9 serum levels served as control. Results were correlated with clinicopathological data. While serum CA19-9 levels were comparable, Olink® analysis confirmed distinct differences between both groups. It revealed significantly elevated levels of factors involved in chemotaxis and migration of immune cells, immune activity, and cell growth in serum of LHMS-patients. Overall, Olink® analysis identified a comprehensive biomarker panel in serum of PDAC patients that could provide the basis for predicting LHMS. However, further studies with larger cohorts are required for its clinical translation.

MiRNAs and isomiRs: Serum-Based Biomarkers for the Development of Intellectual Disability and Autism Spectrum Disorder in Tuberous Sclerosis Complex.

Tuberous sclerosis complex (TSC) is a rare multi-system genetic disorder characterized by a high incidence of epilepsy and neuropsychiatric manifestations known as tuberous-sclerosis-associated neuropsychiatric disorders (TANDs), including autism spectrum disorder (ASD) and intellectual disability (ID). MicroRNAs (miRNAs) are small regulatory non-coding RNAs that regulate the expression of more than 60% of all protein-coding genes in humans and have been reported to be dysregulated in several diseases, including TSC. In the current study, RNA sequencing analysis was performed to define the miRNA and isoform (isomiR) expression patterns in serum. A Receiver Operating Characteristic (ROC) curve analysis was used to identify circulating molecular biomarkers, miRNAs, and isomiRs, able to discriminate the development of neuropsychiatric comorbidity, either ASD, ID, or ASD + ID, in patients with TSC. Part of our bioinformatics predictions was verified with RT-qPCR performed on RNA isolated from patients’ serum. Our results support the notion that circulating miRNAs and isomiRs have the potential to aid standard clinical testing in the early risk assessment of ASD and ID development in TSC patients.

Predictive Biomarkers in the Management of Bladder Cancer: Perspectives in an Evolving Therapeutic Landscape

Bladder cancer (BC) is a heterogeneous disease with prognosis and therapeutic strategies highly dependent on tumor grade and stage. Predictive biomarkers of therapeutic response have been studied to guide selection of intravesical and/or systemic therapy. A predictive biomarker is measured before the start of treatment and provides information on the likelihood of response to a specific therapy. Many candidate predictive biomarkers for BC have been identified, but few have been rigorously validated or distinguished from simply having treatment-agnostic prognostic capacity. Identifying predictive biomarkers tailored to therapeutic mechanism of action has considerable implications for the sequencing of therapies, as well as bladder preservation strategies in advanced disease states. We evaluate predictive tissue-based, urine-based, and serum-based biomarkers across the spectrum of non–muscle-invasive and muscle-invasive BC and preview predictive biomarkers for emerging targeted therapies.