Multiple Serum Research Articles

Prognostic value of plasminogen activator inhibitor-1 in biomarker exploration using multiplex immunoassay in patients with metastatic renal cell carcinoma treated with axitinib.

Background and AimsVascular endothelial growth factor‐directed therapies play a significant role in patients with metastatic renal cell carcinoma (mRCC). Biomarkers for predicting treatment efficacy and resistance are required to develop personalized medicine. We evaluated multiple serum cytokine levels in patients with mRCC treated with axitinib to explore predictive biomarkers.MethodsFrom September 2012 to October 2015, serum samples were collected from 44 patients with mRCC before treatment and 4 weeks after axitinib initiation. Bio‐Plex Pro Human Cancer Biomarker Panels 1 and 2 were used to measure levels of 34 serum biomarkers related to angiogenesis and cell proliferation.ResultsPatients with partial response or stable disease had significantly decreased serum plasminogen activator inhibitor‐1 (PAI‐1) level from pre‐treatment to 4 weeks after axitinib initiation compared with those with progressive disease (P = .022). The median progression‐free survival (PFS) and median overall survival (OS) in patients with increased serum PAI‐1 level from pre‐treatment to 4 weeks after axitinib initiation were significantly shorter than those with decreased serum PAI‐1 level (P = .027 and P = .026, respectively). Increased serum PAI‐1 level from pre‐treatment to 4 weeks after axitinib initiation was an independent prognostic marker for shorter PFS and OS in multivariate analyses (P = .015 and P = .032, respectively). The immunohistochemical staining intensity of PAI‐1 in tumor specimens was significantly associated with Fuhrman grade and presence of distant metastasis (P = .026 and P = .010, respectively).ConclusionsThe initial change in serum PAI‐1 level in the early stage of axitinib treatment could be a useful prognostic biomarker in patients with mRCC.

Extending the provision of cell-free DNA screening for trisomy21.

It is now 12 years since the discovery that fetal cfDNA fragments in maternal plasma can be used to detect Down syndrome. This single marker has a detection rate above 99% and a false-positive rate below 0.2%, a performance greatly exceeding conventional multiple serum and ultrasound marker screening tests. Nevertheless, there has not been widespread replacement of conventional screening by cfDNA. Commercial providers report annual cfDNA testing totaling millions but this covers only a few percent of pregnancies worldwide.

The Evaluation of Some Biomarkers According to Rheumatoid Factor in Early Diagnosis of Rheumatoid Arthritis from Iraqi Patients

Rheumatoid arthritis (RA) was a chronic inflammatory autoimmune disease for long-term that primarily affects small joints and leads to chronic inflammation in synovial. The aimed of the study to identify the relationships among some serological markers (antibodies to citrullinated protein/peptide antigens (ACPAs), anti-mutated citrullinated vimentin (anti-MCV), anti-carbamylated protein (Anti-Carp), anti- heterogeneous nuclear ribonucleoproteins (anti-hnRNP) and Glucose-6-phosphate isomerase (GPI)) and early diagnosis of RA. The study involved (60) Patients of newly diagnosis with RA that divided in to two subgroups (30 RF positive and 30 RF negative) groups and 30 subjects as healthy control group. The serological data from serum concentration of (ACPAs, Anti-MCV, Anti-Carp, Anti-hnRNP, G6PI) estimated by ELISA methods, RF estimated by latex agglutination kits. The results revealed that ACPAs, Anti-MCV, Anti-Carp, Anti-hnRNP, G6PI having a statistical significants at the mean±SD titer different between the two RF groups, a higher value among the refractory RA patients in comparison with the control group. Also effectively distinguishing RA patients groups for RF+ve and RF-ve with showed the sensitivity and specificity of, ACPAs (90.8% , 94.1%, and 88.2%, 86.6%); anti-MCV (66.7% , 33.0% and 70.0%, 70.2%); anti-CarP (76.7% , 90.0% and 93.3%, 78.5%); anti-hnRNP (74.9% , 61.9% and 71.4%, 70.9%) and GPI (77.3% , 76.7% and 84.4%, 80.1%) respectively. This study confirm the importance of measuring multiple serum biomarkers and their com­binations with high diagnostic value for RA and provide sup­port for the early diagnosis of RA.

Associations of multiple serum biomarkers and the risk of cardiovascular disease in China

BackgroundPrevious studies focus on one or several serum biomarkers and the risk of cardiovascular disease (CVD). This study aims to investigate the association of multiple serum biomarkers and the risk of CVD and evaluate the dose-relationship between a single serum metabolite and CVD.MethodsOur case-control study included 161 CVD and 160 non-CVD patients who had a physical examination in the same hospital. We used stratified analysis and cubic restricted analysis to investigate the dose-response relationship of individual serum biomarkers and the CVD incident. Moreover, to investigate serum biomarkers and CVD, we used elastic net regression and logistic regression to build a multi-biomarker model.ResultsIn a single serum biomarker model, we found serum FT4, T4. GLU, CREA, TG and LDL-c were positively associated with CVD. In the male group, serum T4, GLU and LDL-c were positively associated with CVD; and serum TG was positively associated with CVD in the female group. When patients ≤63 years old, serum T4, GLU, CREA and TG were positively associated with CVD, and serum TG and LDL-c were positively associated with CVD when patients > 63 years old. Moreover, serum GLU had nonlinearity relationship with CVD and serum TG and LDL-c had linearity association with CVD. Furthermore, we used elastic regression selecting 5 serum biomarkers (GLU, FT4, TG, HDL-c, LDL-c) which were independently associated with CVD incident and built multi-biomarker model. And the multi-biomarker model had much better sensitivity than single biomarker model.ConclusionThe multi-biomarker model had much higher sensitivity than a single biomarker model for the prediction of CVD. Serum FT4, TG and LDL-c were positively associated with the risk of CVD in single and multiple serum biomarkers models, and serum TG and LDL-c had linearity relationship with CVD.

Quantitative proteomics identifies a plasma multi-protein model for detection of hepatocellular carcinoma

More efficient biomarkers are needed to facilitate the early detection of hepatocellular carcinoma (HCC). We aimed to identify candidate biomarkers for HCC detection by proteomic analysis. First, we performed a global proteomic analysis of 10 paired HCC and non-tumor tissues. Then, we validated the top-ranked proteins by targeted proteomic analyses in another tissue cohort. At last, we used enzyme-linked immunosorbent assays to validate the candidate biomarkers in multiple serum cohorts including HCC cases (HCCs), cirrhosis cases (LCs), and normal controls (NCs). We identified and validated 33 up-regulated proteins in HCC tissues. Among them, eight secretory or membrane proteins were further evaluated in serum, revealing that aldo–keto reductase family 1 member B10 (AKR1B10) and cathepsin A (CTSA) can distinguish HCCs from LCs and NCs. The area under the curves (AUCs) were 0.891 and 0.894 for AKR1B10 and CTSA, respectively, greater than that of alpha-fetoprotein (AFP; 0.831). Notably, combining the three proteins reached an AUC of 0.969, which outperformed AFP alone (P < 0.05). Furthermore, the serum AKR1B10 levels dramatically decreased after surgery. AKR1B10 and CTSA are potential serum biomarkers for HCC detection. The combination of AKR1B10, CTSA, and AFP may improve the HCC diagnostic efficacy.

Effect of perennial dust mites allergy on symptom severity of autumn allergic rhinitis in adults.

Background: Multiple immunoglobulin E (IgE) mediated sensitizations and/or allergies often coexist in patients with allergic rhinitis (AR). Several simultaneous allergen exposures in multiple IgE-mediated sensitizations and/or allergies may increase the allergen load and be related to disease severity. No study has verified whether positive allergen serum IgE levels and allergen categories together are associated with AR severity in adults. Objective: To investigate the effects of perennial dust mites (DMs) allergy and multiple serum sIgE-mediated autumn pollen allergy coexistence on symptom severity in adult patients with AR in autumn. Methods: In total, 153 patients with AR and with autumn pollen allergy (Artemisia argyi, ragweed, and hop) with or without DMs allergy were recruited in the autumn pollen season. Symptom severity was assessed by using the Chinese version of the visual analog scale (VAS): four rhinitis symptoms (sneezing, rhinorrhea, nasal pruritus, and nasal congestion) and two ocular symptoms (ocular itching and/or grittiness and/or redness, and ocular tearing) were scored at approximately the same period. We measured allergen serum sIgE levels for the inhaled allergens. The effects of DMs allergy and multiple autumn pollen allergy coexistence on symptom severity were analyzed. Results: Neither the sum of the autumn pollen allergens categories (total number of positive autumn pollen allergens, i.e., Artemisia argyi or ragweed or hop positive: 1; Artemisia argyi and ragweed positive: 2; Artemisia argyi, ragweed, and hop positive: 3) nor serum sIgE levels( total sIgE levels of positive autumn pollen allergens) exerted any influence on the severity of nasal and ocular symptoms (p > 0.05). When the concomitant DMs allergy status was considered, the sum of the positive autumn pollen allergen categories and accumulated positive autumn pollen and DMs serum sIgE levels (total levels of serum sIgE of positive autumn pollen allergens plus the levels of serum sIgE of DMs) had no influence on patients' symptom severity (p > 0.05). Conclusion: The coexistence of perennial DMs allergy and multiple autumn pollen allergy did not affect the severity of symptoms among adult patients with AR and with autumn pollen allergy in autumn.

Hypothyroidism and its Association with Systemic Lupus Erythematosus: A Cross Sectional Study in Chinese Patients

BackgroundSpecific factors correlated with hypothyroidism in systemic lupus erythematosus (SLE) patients remain unclear. Therefore, we aim to evaluate the prevalence of thyroid dysfunction in Chinese patients with SLE and the relationship between clinical hypothyroidism and SLE. MethodsWe conducted a cross sectional study of the prevalence of thyroid dysfunction in 672 patients with SLE and 605 age- and sex-matched healthy controls. Demographic, clinical, and biochemical data were compared between 58 patients with SLE with hypothyroidism and 197 patients with SLE with euthyroidism. Multivariate analysis was performed using binomial logistic regression analysis. Spearman's rank correlation was used to identify an association between thyroid function and disease activity. ResultsThe prevalence of thyroid dysfunction was significantly higher in patients with SLE than in controls (70.7% vs 19.7%). SLE was associated with higher rates of hypothyroidism (9.6%, P ≤ 0.001) and euthyroid sick syndrome (49.6%, P ≤ 0.001) compared with control subjects. Further analyses showed that hypothyroidism in patients with SLE was associated with high blood pressure, renal disorder, high serum creatinine, high uric acid, hyperlipidaemia, low C3 and C4, positive anti-dsDNA antibodies, and high SLE disease activity index (SLEDAI) score. In multiple logistic regression models, albumin, platelet count, serum creatinine, and anti-dsDNA antibodies were associated with hypothyroidism. Finally, free tri-iodothyronine was significantly negatively correlated with SLEDAI score. ConclusionsHypothyroidism was more prevalent in patients with SLE. There was a relationship between hypothyroidism with renal disorder and lupus activity. Albumin, platelet count, serum creatinine, and anti-dsDNA antibodies were correlated with hypothyroidism.

A 3-Week, Low-Carbohydrate, High-Fat Diet Improves Multiple Serum Inflammatory Markers in Endurance-Trained Males.

Waldman, HS, Heatherly, AJ, Killen, LG, Hollingsworth, A, Koh, Y, and O'Neal, EK. A three-week, low-carbohydrate, high-fat diet improves multiple serum inflammatory markers in endurance-trained males. J Strength Cond Res 36(9): 2502-2508, 2022-This study examined the effects of a low-carbohydrate, high-fat diet (LCHF) on inflammatory marker responses in middle-aged endurance athletes. Eight male runners maintained their habitual mixed diet (HMD) in the first phase of the study before switching to a noncalorically restricted LCHF diet (∼70% of kcals from fat; carbohydrate <50 g) for 3 weeks. Subjects completed a 50-minute fixed pace treadmill running protocol in a hot environment, followed by a 5-km outdoor time trial. Fasting serum samples were collected immediately after exercise and heat stress restriction, and again 24 hours after the exercise/heat stressor. Thirty inflammation markers were assessed using the multiplex flow immunoassay technique. Seven markers (BAFF/TNFSF-13, sCD30/TNFRSF8, sCD163, Chitinase3-like1, gp130SIL-6Rβ, sTNFR-1, and sTNFR-2) reached statistical significance ( p < 0.05) favoring LCHF before exercise, and sCD30/TNFRSF8 favored ( p < 0.05) LCHF before (HMD = 459 ± 111; LCHF = 296 ± 100) and after (HMD = 385 ± 104; LCHF = 285 ± 104 pg·ml -1 ) exercise. Although the current dietary intervention was short in duration, LCHF seems to offer some protection against multiple chronic inflammation markers for physically active men between ages 30 and 50 years.

Abstract NG13: Metabolomic adaptations and correlates of survival to immune checkpoint blockade

Abstract NG13: Metabolomic adaptations and correlates of survival to immune checkpoint blockade

Abstract 4305: Predictive significance of serum proteins on clinical outcome in advanced non-small-cell lung cancer patients treated with immune checkpoint inhibitors

Abstract Background: Though PD-ligand 1 (PD-L1) expression on tumor tissue has been established as companion diagnostics in non-small-cell lung cancer (NSCLC) for anti-PD-1 treatment, additional biomarkers are critically needed. While immune related adverse events (irAEs) have been reported to be associated with efficacy, there are no established biomarkers to predict irAEs onset. Here, we measured multiple serum proteins in NSCLC patients treated with immune checkpoint inhibitors (ICIs) and explored the potential of predicting clinical response and irAE onset. Patients and Methods: Advanced NSCLC patients received nivolumab, pembrolizumab or atezolizumab until progressive disease (PD) or unacceptable toxicity. Serum samples were collected at baseline and after the treatment (at week 4 or week 6). Durable clinical benefit (DCB) was defined as patients whose response was lasting over 6 months. Using Luminex technology, serum levels of 41 proteins consisting of cytokines, chemokines, growth factors, and angiogenesis factors were measured. All statistical analyses were carried out using JMP Pro software (ver. 14.0). Cut-off value of serum protein levels were estimated with receiver operating characteristic curve. Results: Ninety-eight patients were registered in the study between January 2016 and May 2019 at Wakayama Medical University Hospital and 97 were included in the final analysis. Demographics of the patients were as follows: median age 70 (range, 49 to 91); male 78%; stage III/IV 31/69%; squamous/non-squamous/unknown 29/69/2%; previous treatment: 0/≥1 22/78%; ICI: nivolumab/pembrolizumab/atezolizumab 46/41/13%. Objective response rate was 24.7% (24/97), and disease control rate was 46.4% (45/97). Among 41 serum proteins measured serially, IL-8 levels at baseline were significantly lower in DCB patients than non-DCB patients in entire cohort (p &amp;lt; 0.05). Among those, Kaplan-Meier analysis revealed lower levels of IL-8 were significantly associated with longer PFS (cut-off, 11.9217 pg/ml; median PFS, 293 vs 59 days; p &amp;lt; 0.01). In second or later line subset, GRO-α, IL-8, IP-10, PLGF and VEGF-D were significantly lower in DCB patients than non-DCB-patients at baseline (p &amp;lt; 0.05). Lower levels of those proteins except for VEGF-D were significantly associated with longer PFS ((cut-off, 1.073 ng/ml, 11.9217 pg/ml, 480.8 pg/ml, and 14.83 pg/ml; median PFS, 183 vs 49, 293 vs 49, 133 vs 42, and 153 vs 49 days; 95% CI, 56-322 days, 69 days-not reached, 58-211 days and 52-293 days; p &amp;lt; 0.05, 0.01, 0.05, 0.01). Follistatin which we previously reported to be associated with DCB upon novolumab treatment, also showed significant association with longer PFS both in entire cohort (cut-off, 572.5; median PFS, 160 vs 59 days; 95% CI, 68-322 days, p &amp;lt; 0.05) and 2nd or later line subset (cut-off, 572.5; median PFS, 100 vs 49 days; 95% CI, 39-322 days, p &amp;lt; 0.05), though it was not significantly different between DCB and non-DCB. Regarding irAEs, GRO-α levels were significantly increased between baseline and the second sampling point in patients who acquired irAEs both in entire cohort and 2nd or later line subset (p &amp;lt; 0.01). Conclusions: We identified potential serum protein markers associated with clinical benefit and irAEs in advanced NSCLC treated with ICIs by multiplex protein-based assay. Citation Format: Jun Oyanagi, Yasuhiro Koh, Koichi Sato, Shunsuke Teraoka, Atsushi Hayata, Nahomi Tokudome, Hiroaki Akamatsu, Yuichi Ozawa, Keiichiro Akamatsu, Masanori Nakanishi, Hiroki Ueda, Nobuyuki Yamamoto. Predictive significance of serum proteins on clinical outcome in advanced non-small-cell lung cancer patients treated with immune checkpoint inhibitors [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4305.

Prognosis of Macrophage Density in the Absence of Neutrophils in Differentiated Thyroid Cancer

BackgroundDespite the advances in treatment of differentiated thyroid cancer (DTC), predicting prognosis remains a challenge. Immune cells in the tumor microenvironment may provide an insight to predicting recurrence. Therefore, the objective of this study was to investigate the association of tumor-associated macrophages (TAMs) and tumor-associated neutrophils (TANs) with recurrence in DTC and to identify serum cytokines that correlate with the presence of these immune cells in the tumor. Materials and methodsForty-two DTC tissues from our institutional neoplasia repository were stained for immunohistochemistry markers for TAMs and TANs. In addition, cytokine levels were analyzed from these patients from preoperative blood samples. TAM and TAN staining were compared with clinical data and serum cytokine levels. ResultsNeither TAM nor TAN scores alone correlated with tumor size, the presence of lymph node metastases, multifocal tumors, lymphovascular or capsular invasion, or the presence of BRAFV600E mutation (all P > 0.05). There was no association with recurrence-free survival (RFS) in TAN density (mean RFS, 169.1 versus 148.1 mo, P = 0.23) or TAM density alone (mean RFS, 121.3 versus 205.2 mo, P = 0.54). However, when scoring from both markers were combined, patients with high TAM density and TAN negative scores had significantly lower RFS (mean RFS, 50.7 versus 187.3 mo, P = 0.04) compared with the remaining cohort. Patients with high TAM/negative TAN tumors had significantly lower serum levels of interleukin 12p70, interleukin 8, tumor necrosis factor alpha, and tumor necrosis factor beta. ConclusionsIn DTCs, high density of TAMs in the absence of TANs is associated with worse outcome. Assessment of multiple immune cell types and serum cytokines may predict outcomes in DTC.

Reusable optofluidic point-of-care testing platform with lyophilized specific antibody for fluorescence detection of cholylglycine in serum.

A reusable optofluidic point-of-care testing platform (OPOCT) was successfully constructed through integrating evanescent wave fluorescence technology into an all-fiber-based optofluidic system. The compact design of the OPOCT allows it to be portable and suitable for on-site sensitive determination of biomarkers in serum without complicated and costly procedures. The sensitivity of 90.9pM for antibody determination is observed thanks to the high transmission efficiency of excitation light and fluorescence in the OPOCT. The affinity constant between cholylglycine (CG) and anti-CG antibody was quantified using this platform based on the proposed theory. Using the lyophilized fluorescence-labeled specific antibody and reusable fiber optic biosensor, the OPOCT is applied to the one-step sensitive determination of CG in serum, which eliminates the dearth associated with liquid reagent handling, disposable biosensors, and user intervention. A limit of detection of 0.025μg/mL for CG is obtained, which is far more than adequate for meeting diagnostic requirements. The matrix effect of serum samples on the evanescent wave-based optofluidic biosensor can be effectively reduced by simple dilution of serum samples. The performance of the OPOCT also compared favorably with that of a commercial turbidimetric inhibition immunoassay through analyzing multiple serum samples. This platform is ready to expand to measure any other biomarker by using its specific antibody. The simplicity, sensitivity, cost-effectiveness, and robustness of the OPOCT enable the early diagnosis of disease and making a timely clinical decision. Graphical abstract .

Abstract PR02: Glyphosate as a potential carcinogen for multiple myeloma

Abstract Glyphosate is the most commercially successful and the most widely used herbicide in the United States and worldwide. It represents about 50% of United States herbicide usage at approximately 250 million pounds annually; the annual use of glyphosate worldwide is about 1.8 billion pounds. There has been considerable debate around the carcinogenicity of glyphosate. IARC labeled glyphosate as “probably carcinogenic to humans.” However, other agencies like the EPA have maintained that glyphosate is unlikely to pose a carcinogenic risk. In epidemiologic studies on the association of cancer and glyphosate, only multiple myeloma (MM) and non-Hodgkin lymphoma (NHL) have a positive and statistically significant association with glyphosate. MM is a plasma cell neoplasm and the second most common hematologic malignancy in the United States. Virtually all MM cases are preceded by a precursor condition, monoclonal gammopathy of undetermined significance (MGUS). MM is regarded as a disease with environmental causes, yet its etiology and what prompts progression remain largely unclear. We employed the Vk*Myc mouse model of MM to assess the impact of glyphosate exposure on MM pathogenesis. In both wild-type and Vk*Myc mice, glyphosate induced splenomegaly and multiple peripheral blood abnormalities, including anemia and high serum IgG levels, hallmarks of MGUS and MM in humans. In Vk*Myc mice, glyphosate triggered bone lytic lesions, as well as renal damage, a phenomenon associated with human MM kidney disease. Most importantly, glyphosate upregulated Aicda, which encodes the activation-induced cytidine deaminase (AID) in the spleen and bone marrow of both wild-type and Vk*MYC mice. AID mediates somatic hypermutation and class-switch recombination of immunoglobulin genes in B cells to produce diverse immunoglobins in secretive forms. Our data indicate that glyphosate accelerates MGUS development and promotes progression to MM in genetically predisposed mice. Given that AID dysregulation is a key pathogenic driver in both MM and NHL, this work offers the first direct experimental evidence establishing glyphosate as an environmental risk factor for MM and NHL. This abstract is also being presented as Poster A50. Citation Format: Yong Li. Glyphosate as a potential carcinogen for multiple myeloma [abstract]. In: Proceedings of the AACR Special Conference on Environmental Carcinogenesis: Potential Pathway to Cancer Prevention; 2019 Jun 22-24; Charlotte, NC. Philadelphia (PA): AACR; Can Prev Res 2020;13(7 Suppl): Abstract nr PR02.

Applying Serum Proteins and MicroRNA as Novel Biomarkers for Early-Stage Cervical Cancer Detection

Recently, we have been seeing emerging applications of non-invasive approaches using serum biomarkers including miRNA and proteins in detection of multiple cancers. Currently, majority of these methods only use solitary type of biomarkers, which often lead to non-satisfactory sensitivity and specificity in clinical applications. To this end, we established a unique biomarker panel in this study, which determined both squamous cell carcinoma antigen (SCC Ag) degree and miRNA-29a, miRNA-25, miRNA-486-5p levels in blood for detection of early-stage cervical cancer. We designed our study with two phases: a biomarker discovery phase, followed by an independent validation phase. In total of 140 early-stage cervical cancer patients (i.e., AJCC stage I and II) and 140 healthy controls recruited in the biomarker discovery phase, we achieved sensitivity of 88.6% and specificity of 92.9%. To further assess the predictive power of our panel, we used it to an independent patient cohort that consisted of 60 early-stage cervical cancer individuals as well as 60 healthy controls, and successfully achieved both high sensitivity (80.0%) and high specificity (96.7%). Our study indicated combining analyses of multiple serum biomarkers could improve the accuracy of non-invasive detection of early-stage cervical cancer, and potentially serve as a new liquid biopsy approach for detecting early-stage cervical cancer.

Relationship between multiple healthy lifestyles and serum lipids among adults in rural China: A population-based cross-sectional study

Although several single lifestyles have been reported to meliorate serum lipid levels, there is little evidence of the relationship between multiple healthy lifestyles and lipid levels in Chinese adults. Cross-sectional data of 35,150 participants aged 18–79 years without dyslipidemia at baseline in the Henan Rural Cohort were collected from 2015 to 2017, to investigate the associations between individual and combinations of 6 healthy lifestyle factors and lipid levels. In multivariate linear regression analyses, non-current smoking, non-current alcohol consumption, regular physical exercise, healthy diet, lower body mass index, and lower waist-to-hip ratio were significantly associated with lower concentration of total cholesterol (TC), triglyceride (TG), and low-density lipoprotein cholesterol (LDL-C) and higher concentrations of high-density lipoprotein cholesterol (HDL-C) (all P < .001). There were positive quantitative correlations between the number of healthy lifestyle factors and the low levels of lipids (all P for linear trend <0.001). People with 6 healthy lifestyle factors had 0.72-mmol/L (95% confidence interval [CI]: 0.64 to 0.81) lower TC, 1.02-mmol/L (95% CI: 0.93 to 1.11) lower TG, 0.54-mmol/L (95% CI: 0.47 to 0.61) lower LDL-C, and 0.32-mmol/L (95% CI: 0.29 to 0.34) higher HDL-C, than people who had 0–1 healthy lifestyle factors. In this study, we found an association between an increased number of healthy lifestyle factors and better serum lipid profiles. The causality and temporality between maintenance of a healthy lifestyle and optimal lipid levels merit further investigations.

Predicting the efficacy of nivolumab combined with radiation therapy by longitudinal liquid biopsy with artificial intelligence for patients with metastatic breast cancer (translational research of the KBCRN-B-002 trial).

e15046 Background: Advances in immuno-oncology (IO), i.e., programmed cell-death protein 1 (PD-1) inhibitors, are revolutionizing cancer treatment by offering hope for a cure. However, the benefit of IO agents in metastatic breast cancer (MBC) remains limited. For immunotherapy to be successful, it is essential to understand the tumor immune contexture. Systemic immune alterations, such as the signature of peripheral blood mononuclear cells (PBMC) and cytokine expression, are one of the hallmarks of successful immunotherapy in MBC. The abscopal effect induced by radiation therapy (RT) is known to be a modulator of systemic immune alterations, and the abscopal effect is considered as a systemic anti-tumor immune response. In this study, we integrated time series multi-omics data in order to identify alteration of the immune signatures after RT combined with nivolumab, a PD-1 inhibitor, using a longitudinal liquid biopsy with artificial intelligence. Methods: This study was conducted as translational research, part of the KBCRN-B-002 trial, which is a multicenter phase Ib/II study evaluating the safety and efficacy of nivolumab in combination with RT in patients with HER2-negative MBC (UMIN: UMIN000026046). Twenty-nine patients were included in the translational analysis set. The multi-omics data included data from RNAseq, mass cytometry (CyTOF) and multiple cytokines, using PBMC, serum and plasma. We collected time series data, which covered the baseline, two weeks after starting therapy, four weeks after starting therapy and the timing of progressive disease. For integrated analysis of the multi-omics data, a machine-learning method was developed. Results: Our integrated analysis (involving a longitudinal liquid biopsy and machine-learning method) identified an apparent cluster of responder groups and alteration of the systemic immune signatures. Single-cell analysis using CyTOF results including the signature of the immune cell composition, effector immune cells and immune suppressor cells. Conclusions: Our method clustered responders well and identified candidates of the systemic immune signatures for actionable hallmarks of RT combined with IO agent by a longitudinal liquid biopsy. Clinical trial information: NCT03430479 .

Landscape of circulating diagnostic biomarkers in pancreatic malignancies

: Pancreatic cancer remains the most lethal adult cancer in the US that is in large part due to the lack of effective early detection biomarkers. Although a variety of blood based technologies have been developed, each of these assays have different advantages and disadvantages. Here, we will review the landscape of blood based biomarkers in the context of pancreatic cancer early detection. Tumor biomarkers (CA19-9, CEA) are widely used, but often have limited specificity to cancer, and panel-based approaches combining multiple serum proteins may prove more promising in early detection of pancreatic cancer. Metabolic markers, including the substrates, intermediates, and by-products of aberrant pathways can also be quantified and used as biomarkers, particularly since pancreatic cancer almost universally exploits KRAS mutations leading aberration activation of the MEK/ERK/AKT pathways. More recent approaches have also attempted to use cell-free DNA released from tumor cells into the bloodstream to directly sequence and identify mutations and serve as a ‘liquid biopsy.’ Alternatively, approaches are being used to capture rare circulating epithelial cells that have entered into the bloodstream (circulating tumor cells, or CTCs) even in pre-neoplastic lesions. Extracellular vesicles (EVs) found in plasma are a new emerging source of tumor enriched cargo (DNA, RNA, proteins) providing a multi-prong approach towards early tumor detection. Although many of these approaches are promising, they need to be validated in large-scale human trials using appropriate controls. Altogether, the complementary performance characteristics of different biomarkers point to a combination of these technologies as the future of a pancreatic cancer early detection liquid biopsy.

Evaluation of maternal serum biomarkers in predicting outcome of successful expectant management of tubal ectopic pregnancies

ObjectiveTo assess the value of multiple serum biomarkers for the prediction of successful outcome of expectant management in women with tubal ectopic pregnancy (TEP). Study designWomen with a conclusive ultrasound diagnosis of TEP had a blood test to measure β-human chorionic gonadotropin (β-hCG), progesterone, inhibin A, activin A and high sensitivity C-reactive protein (hsCRP) at the initial visit. Women presenting with pain, serum β-hCG ≥ 1500 IU, evidence of a live ectopic pregnancy or a significant haemoperitoneum were advised to have emergency surgery. Women eligible for expectant management were followed-up prospectively until serum β-hCG declined to non-pregnant level or surgical treatment was required. ResultsA total of 93 women with a TEP were included in the final cohort. Emergency surgery was carried out in 42/93 (45 %) of women whilst 51/93 (55 %) were managed expectantly. Of the latter group, 42/51 (82 %) had successful expectant management and 9/51(18 %) required surgical procedure after a period of follow up. On multi-variable analysis, only higher values of serum β-hCG and progesterone at the initial visit were associated with a lower chance of successful expectant management of TEP. A one-unit increase in either variable on the log-scale was associated with an approximate 20-fold reduction in the odds of a successful outcome. Conclusion(s)Serum β-hCG and progesterone were significantly lower in women who had successful expectant management of TEP. Other biomarkers under consideration were not significantly different in women with successful and failed expectant management.

Abstract 29: Metabolomic Signatures of Sedentary Behavior and Cardiometabolic Traits in US Hispanics/Latinos: Hispanic Community Health Study / Study of Latinos (HCHS/SOL)

Introduction: Sedentary behavior is associated with cardiometabolic risk independent of physical activity, but the underlying mechanisms are not fully understood. Hypothesis: Sedentary behavior may influence serum metabolite profiles which are associated with cardiometabolic risk factors. Methods: The analyses included 2711 participants (43% men) of the HCHS/SOL aged 18 to 74 years, free of CVD and other major chronic diseases. Serum metabolomics was performed using an untargeted approach. Physical activity and sedentary behavior were measured by accelerometer (Actical) for a 7-day period. Survey linear regressions were used to examine associations between sedentary behavior and metabolites. Metabolite scores were calculated by summing up z-scores of significant metabolites within the same sub-pathway. Results: Of 624 known metabolites, 95 were significantly associated with sedentary behavior ( P &lt;0.05 after FDR correction), adjusting for socioeconomic and lifestyle factors including moderate to vigorous physical activity ( Figure 1A ). The majority of significant metabolites were clustered into 7 sub-pathways. Three sub-pathway scores (branched-chain amino acids [BCAA], acylcarnitine and medium-chain fatty acid metabolism) were positively associated with sedentary behavior, while the other 4 sub-pathway scores (acylcholine, plasmalogen, nicotinate, and arginine) were inversely associated with sedentary behavior. Among these scores, the BCAA score showed unfavorable correlations with all cardiometabolic traits including obesity measures, glycemic traits, blood lipids and blood pressure. The acylcholine score showed favorable correlations with obesity measures and glycemic traits, though positive correlations with blood lipids were observed ( Figure 1B) . Conclusions: Our study identified multiple serum metabolites associated with sedentary behavior independent of physical activity, suggesting potential mechanistic links between sedentary behavior and cardiometabolic diseases.

Human Pluripotent Stem Cell-Derived Hepatocytes Show Higher Transcriptional Correlation with Adult Liver Tissue than with Fetal Liver Tissue.

Human pluripotent stem cell-derived hepatocytes (hPSC-HEP) display many properties of mature hepatocytes, including expression of important genes of the drug metabolizing machinery, glycogen storage, and production of multiple serum proteins. To this date, hPSC-HEP do not, however, fully recapitulate the complete functionality of in vivo mature hepatocytes. In this study, we applied versatile bioinformatic algorithms, including functional annotation and pathway enrichment analyses, transcription factor binding-site enrichment, and similarity and correlation analyses, to datasets collected from different stages during hPSC-HEP differentiation and compared these to developmental stages and tissues from fetal and adult human liver. Our results demonstrate a high level of similarity between the in vitro differentiation of hPSC-HEP and in vivo hepatogenesis. Importantly, the transcriptional correlation of hPSC-HEP with adult liver (AL) tissues was higher than with fetal liver (FL) tissues (0.83 and 0.70, respectively). Functional data revealed mature features of hPSC-HEP including cytochrome P450 enzymes activities and albumin secretion. Moreover, hPSC-HEP showed expression of many genes involved in drug absorption, distribution, metabolism, and excretion. Despite the high similarities observed, we identified differences of specific pathways and regulatory players by analyzing the gene expression between hPSC-HEP and AL. These findings will aid future intervention and improvement of in vitro hepatocyte differentiation protocol in order to generate hepatocytes displaying the complete functionality of mature hepatocytes. Finally, on the transcriptional level, our results show stronger correlation and higher similarity of hPSC-HEP to AL than to FL. In addition, potential targets for further functional improvement of hPSC-HEP were also identified.