CHI3L1 Research Articles

Construction of a diagnostic nomogram model for lymph node metastasis in gastric cancer based on human cartilage glycoprotein-39, carbohydrate antigen 50, and serum ferritin

Construction of a diagnostic nomogram model for lymph node metastasis in gastric cancer based on human cartilage glycoprotein-39, carbohydrate antigen 50, and serum ferritin

Comparison of Antigen Retrieval Methods for Immunohistochemical Analysis of Cartilage Matrix Glycoproteins Using Cartilage Intermediate Layer Protein 2 (CILP-2) as an Example.

The aim of this study was to compare different antigen retrieval methods to improve the outcome of immunohistochemistry (IHC) performed on osteoarthritic (OA) cartilage obtained from total knee replacement operation. A voluminous and dense extracellular matrix of articular cartilage inhibits antibody penetration, and therefore, proteins present at low concentrations and masked during fixation may need antigen retrieval to enhance an IHC outcome. We focused on the IHC detection of a minor but diagnostically promising cartilage glycoprotein, CILP-2 (cartilage intermediate layer protein 2), to demonstrate the effect of four different protocols: (1) heat-induced epitope retrieval (HIER), (2) proteolytic-induced epitope retrieval applying proteinase K and hyaluronidase (PIER), (3) HIER combined with PIER, and (4) no antigen retrieval (control). A semi-quantitative staining assessment based on the CILP-2 staining extent was applied. Out of the tested antigen retrieval protocols, the best CILP-2 IHC staining results were achieved by PIER. Combining PIER with HIER did not improve CILP-2 staining in the given experimental setting. Rather the opposite, the application of heat reduced the positive effect of PIER on CILP-2 staining and resulted in the frequent detachment of sections from the slides. Our findings emphasize the need for proper adaptation of antigen retrieval protocols for IHC to maximize the quantitative evaluation of minor matrix proteins in OA articular cartilage samples.

Alcohol drinking modified the effect of plasma YKL-40 levels on stroke-specific mortality of acute ischemic stroke

ObjectivesOur study aimed to evaluate the association between plasma human cartilage glycoprotein-39 (YKL-40) and stroke-specific mortality at two years in acute ischemic stroke patients according to the drinking status and amount of alcohol consumption. We further investigated the effect of the interaction between these conditions and YKL-40 levels on the outcome. MethodsWe measured plasma YKL-40 levels in 3267 participants from the China Antihypertensive Trial in Acute Ischemic Stroke. Outcome data on stroke-specific mortality were collected at two years after stroke onset. ResultsDuring the two years of follow-up, 208 (6.4 %) patients, including 44 drinkers and 164 nondrinkers, died of stroke-specific causes. The patients in the highest quartile of YKL-40 had a 3.52-fold (95 % CI: 1.15–10.76, P for trend = 0.006) risk of stroke-specific mortality compared with those in the lowest quartile among drinkers. However, no significant association between YKL-40 and the outcome was observed among nondrinkers (HR: 1.18, 95 % CI: 0.75–1.86, P for trend = 0.08). Alcohol drinking modified the effect of YKL-40 on the outcome (P for interaction = 0.04). Subgroup analyses revealed that each 1-unit increase in log-transformed YKL-40 was associated with a 72 % greater risk of stroke-specific mortality for light drinkers. This association was amplified with a 226 % increased risk of the outcome among heavy drinkers. ConclusionsElevated YKL-40 levels were associated with an increased risk of stroke-specific mortality at two years among drinkers with ischemic stroke. Drinking status substantially modified the effect of plasma YKL-40 levels on the outcome. This effect was amplified with the increased amount of alcohol consumption.Registration: URL: https://www.clinicaltrials.gov; Unique identifier: NCT01840072.

Evolutionary insights into sequence modifications governing chitin recognition and chitinase inactivity in YKL-40 (HC-gp39, CHI3L1)

YKL-40, also known as human cartilage glycoprotein-39 (HC-gp39) or CHI3L1, shares structural similarities with chitotriosidase (CHIT1), an active chitinase, but lacks chitinase activity. Despite being a biomarker for inflammatory disorders and cancer, the reasons for YKL-40's inert chitinase function have remained elusive. This study reveals that the loss of chitinase activity in YKL-40 has risen from multiple sequence modifications influencing its chitin affinity. Contrary to the common belief associating the lack of chitinase activity with amino acid substitutions in the catalytic motif, attempts to activate YKL-40 by creating two amino acid mutations in the catalytic motif (MT-YKL-40) proved ineffective. Subsequent exploration that included creating chimeras of MT-YKL-40 and CHIT1 catalytic domains (CatD) identified key exons responsible for YKL-40 inactivation. Introducing YKL-40 exons 3, 6, or 8 into CHIT1 CatD resulted in chitinase inactivation. Conversely, incorporating CHIT1 exons 3, 6, and 8 into MT-YKL-40 led to its activation. Our recombinant proteins exhibited properly formed disulfide bonds, affirming a defined structure in active molecules. Biochemical and evolutionary analysis indicated that the reduced chitinase activity of MT-YKL-40 correlates with specific amino acids in exon 3. M61I and T69W substitutions in CHIT1 CatD diminished chitinase activity and increased chitin binding. Conversely, substituting I61 with M and W69 with T in MT-YKL-40 triggered chitinase activity while reducing the chitin-binding activity. Thus, W69 plays a crucial role in a unique subsite within YKL-40. These findings emphasize that YKL-40, though retaining the structural framework of a mammalian chitinase, has evolved to recognize chitin while surrendering chitinase activity.

A clinically applicable connectivity signature for glioblastoma includes the tumor network driver CHI3L1

Tumor microtubes (TMs) connect glioma cells to a network with considerable relevance for tumor progression and therapy resistance. However, the determination of TM-interconnectivity in individual tumors is challenging and the impact on patient survival unresolved. Here, we establish a connectivity signature from single-cell RNA-sequenced (scRNA-Seq) xenografted primary glioblastoma (GB) cells using a dye uptake methodology, and validate it with recording of cellular calcium epochs and clinical correlations. Astrocyte-like and mesenchymal-like GB cells have the highest connectivity signature scores in scRNA-sequenced patient-derived xenografts and patient samples. In large GB cohorts, TM-network connectivity correlates with the mesenchymal subtype and dismal patient survival. CHI3L1 gene expression serves as a robust molecular marker of connectivity and functionally influences TM networks. The connectivity signature allows insights into brain tumor biology, provides a proof-of-principle that tumor cell TM-connectivity is relevant for patients’ prognosis, and serves as a robust prognostic biomarker.

Serum neopterin, YKL-40, IL-6, and TNF-α in patients with obstructive sleep apnea

Objective: Obstructive Sleep Apnea (OSA) is characterized by recurrent episodes of complete or partial upper airway obstruction during sleep. The role of locally increased inflammation in the upper respiratory tract is known in the pathogenesis of OSA. This study aimed to examine the role of some inflammatory markers in OSA. Materials and Methods: Adult patients who underwent diagnostic polysomnography (PSG) for the first time, were evaluated in this prospective study. Serum levels of neopterin, human cartilage glycoprotein 39 (YKL-40), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α) were examined in the study group. Results: The study included 165 adult patients. Individuals of 56 who were did not have OSA were taken as the control group. Patients diagnosed with OSA (n=109) were included in the study and divided into two groups mild-moderate and severe based on the OSA apnea-hypopnea index (AHI). The study groups were divided into 3 cathegories: AHI <5 (control) Group 1, AHI 5-15 (mild), and AHI 15-30 (moderate) Group 2, AHI>30 (severe) Group 3. Individuals of 165 who completed the study [Group 1 (n=56), Group 2 (n=69), Group 3 (n=40)] were included in the statistical evaluation. There was a significant difference between the groups in terms of neopterin, YKL-40, and IL-6 values (p<0.05). The median value of IL-6 and neopterin measured in the severe group was significantly higher than the value measured in the control and mild-moderate groups (p<0.001). The mean value of YKL-40 measured in the severe group was significantly higher than the value measured in the control group (p=0.012). No significant difference was observed in TNF-α serum levels of groups. Conclusion: Serum levels of neopterin, YKL-40, and IL-6 were found to be elevated in OSA patients (especially in the severe group).

Increased Circulating Levels of YKL-40 Protein in Children and Adolescents with Hemophilia: Correlation with the Degree of Hemophilic Arthropathy

Introduction: Hemophilia A and B are rare X-linked inherited bleeding disorders caused by complete or partial deficiency of coagulation factors VIII and IX, respectively. Recurrent joint bleeding (hemarthrosis) is the most frequent clinical manifestation of severe hemophilia. Unless appropriately managed, even subclinical hemarthrosis can lead to hemophilic arthropathy, that severely affects the quality of life of patients. Chitinase-3-like protein 1 (CHI3L1), also known as YKL-40, is a 40-kDa heparin-human cartilage glycoprotein without enzymatic activity, that is encoded by the CHI3L1 gene in humans and its name derives from the one letter code for the three terminal amino acids - tyrosine (Y), lysine (K) and leucine (L). YKL-40 is secreted in vitro and in vivo in the arthritic joint by various cell types, such as activated macrophages, chondrocytes, synoviocytes and neutrophils. Actually, it is a major protein that has been identified in the chondrocytes from arthritic knee joints.In fact, synovial as well as serum YKL-40 levels are found increased in joint diseases like rheumatoid arthritis and osteoarthritis, indicating its potential role as a biomarker of inflammation and tissue remodeling or degradation, whereas it is absent in normal joints. In our knowledge, there are not any reports linking YKL-40 expression with hemophilic arthropathy, thus we aimed to investigate if this protein could be implicated in hemophilic arthropathy severity, by measuring its circulating levels and correlate them with hemophilic arthropathy features and laboratory parameters in children and adolescents with hemophilia A and B. Patients and Methods: Eighty children and adolescents (median age:14 years, range 10-18 years) with severe (n= 60), moderate (n= 8) and mild (n= 12) hemophilia A or B (n= 7), on prophylaxis or on demand treatment (n= 12) were included in the study, while 35 apparently healthy male individuals matched for age served as controls. No signs of hemophilic arthropathy (NHAP), i.e. Hemophilia Joint Health Score 2.1 ≤ 2 and Hemophilia Early Detection Arthropathy with Ultrasound Score ≤ 1, were detected in 34/80 patients with median age of 13.5 years. In 37/80 patients with median age of 14.25 years arthropathy (HAP) was confirmed, while 18/80 patients with median age of 15.75 years presented with acute bleeding (AB)-hemarthrosis or muscle bleed- at the time of the laboratory evaluation. In patients and controls YKL-40 concentrations were determined by means of an immunoenzymatic technique. Results: We found that: a) Circulating YKL-40 levels in patients with hemophilia were significantly higher compared to controls, 21.9±2.3 ng/mL (95% Confidence Intervals of the mean (CI): 19.6;24.2 ng/mL) vs 17.0±2.5 ng/mL (CI: 14.5;19.4 ng/mL), respectively, p=0.003. b) Patients with arthropathy (HAP) tend to express higher circulating YKL-40 levels (25.5±2.0 ng/mL) than patients without arthropathy (NHAP)(20.1±1.8 ng/mL) and patients with acute bleeding (AB) (19.7±2.4 ng/mL), respectively, p-trend=0.06. Conclusions: To the best of our knowledge, this is the first study to evaluate the potential clinical significance of circulating YKL-40 levels in patients with hemophilia and demonstrated that it could reflect hemophilic arthropathy activity in clinical or even in subclinical level. As YKL-40 in synovial fluid is derived from cells in the inflamed/damaged synovium, such as chondrocytes, we postulate that it may be involved in the pathophysiology of the hemophilic arthropathy processes and reflect local disease activity. Further studies are essential on the role of YKL-40 in the evolution of hemophilic arthropathy, in order to fully elucidate its clinical significance and prognostic utility.

YKL-40 Promotes the Expression of Inflammatory Factors in Type Ⅱ Alveolar Epithelial Cell Model of A549 Cell Line

YKL-40, also known as chitinase-3-like-1 (CHI3L1), is a human cartilage glycoprotein-39, with its N-terminus consisting of tyrosine (Y), lysine (K), and leucine (L), hence the name YKL-40. In this study, we explored whether YKL-40 could promote the expression of inflammatory factors in type Ⅱ alveolar epithelial cells. A549 cells were cultured in vitro with interleukin (IL)-1β (20 ng/mL), IL-6 (20 ng/mL), tumor necrosis factor-alpha (TNF-α) (20 ng/mL), and interferon-gamma (IFN-γ) (20 ng/mL). The expression of YKL-40 transcription was determined by RT-qPCR. A549 cells were cultured with IL-1β at 5, 10, and 20 ng/mL and the expression of YKL-40 protein was determined by Western blot. A549 cells were cultured with recombinant YKL-40 protein at 0, 100, 500, and 1 000 ng/mL and the expression levels of IL-6 and IL-8 were measured by RT-qPCR. Three pairs of small interfering RNAs targeting YKL-40 (si- YKL-40-1/2/3) and the negative control (NC) were designed and used to transfect A549 cells, respectively, and the expression of YKL-40 was determined by RT-qPCR and Western blot. si- YKL-40-3 was screened out for subsequent experiments. In A549 cells, si- YKL-40-3 and si-NC were transfected and, then, IL-1β (20 ng/mL) was added in for culturing. The expression of YKL-40, IL-6, and IL-8 was determined by RT-qPCR and the expression of multiple factors in the supernatant was measured with the QAH-INF-1 kit. RT-qPCR results showed that IL-1β could up-regulate YKL-40 protein transcription level compared with that of the control group and the difference was statistically significant ( P<0.01), but IL-6, TNF-α, and IFN-γ could not up-regulate YKL-40 protein transcription level. Western blot results showed that IL-1β (20 ng/mL) could significantly promote the expression of YKL-40 and, compared with that of the control group, the differences showed by groups treated with different concentrations of IL-1β were all statistical significant ( P<0.01). After adding human recombinant YKL-40 protein to A549 cells, the results showed that the expression of inflammatory factors IL-6 and IL-8 was significantly increased and the difference was statistically significant compared with that of the control group ( P<0.05). After the expression of YKL-40 was decreased by si- YKL-40-3 transfection, the expression of IL-6 ( P<0.05), IL-8 ( P<0.05), and other inflammatory factors was inhibited compared with that of the control group. YKL-40 can promote the expression and secretion of IL-6, IL-8, and other acute inflammatory factors in A549 cell line, a type Ⅱ alveolar epithelial cell model, thus aggravating the inflammatory response. Targeted inhibition of YKL-40 expression may effectively inhibit inflammatory response.

Plasma Biomarkers and Incident CKD Among Individuals Without Diabetes

Biomarkers of kidney disease progression have been identified in individuals with diabetes and underlying chronic kidney disease (CKD). Whether or not these markers are associated with the development of CKD in a general population without diabetes or CKD is not well established. Prospective observational cohort. In the Atherosclerosis Risk in Communities) study, 948 participants were studied. The baseline plasma biomarkers of kidney injury molecule-1 (KIM-1), monocyte chemoattractant protein-1 (MCP-1), soluble urokinase plasminogen activator receptor (suPAR), tumor necrosis factor receptor 1 (TNFR-1), tumor necrosis factor receptor 2 (TNFR-2), and human cartilage glycoprotein-39 (YKL-40) measured in 1996-1998. Incident CKD after 15 years of follow-up defined as≥40% estimated glomerular filtration rate decline to<60mL/min/1.73m2 or dialysis dependence through United States Renal Data System linkage. Logistic regression and C statistics. There were 523 cases of incident CKD. Compared with a random sample of 425 controls, there were greater odds of incident CKD per 2-fold higher concentration of KIM-1 (OR, 1.49; 95% CI, 1.25-1.78), suPAR (OR, 2.57; 95% CI, 1.74-3.84), TNFR-1 (OR, 2.20; 95% CI, 1.58-3.09), TNFR-2 (OR, 2.03; 95% CI, 1.37-3.04). After adjustment for all biomarkers, KIM-1 (OR, 1.42; 95% CI, 1.19-1.71), and suPAR (OR, 1.86; 95% CI, 1.18-2.92) remained associated with incident CKD. Compared with traditional risk factors, the addition of all 6 biomarkers improved the C statistic from 0.695-0.731 (P<0.01) and using the observed risk of 12% for incident CKD, the predicted risk gradient changed from 5%-40% (for the 1st-5th quintile) to 4%-44%. Biomarkers and creatinine were measured at one time point. Higher levels of KIM-1, suPAR, TNFR-1, and TNFR-2 were associated with higher odds of incident CKD among individuals without diabetes. For people with diabetes or kidney disease, several biomarkers have been shown to be associated with worsening kidney disease. Whether these biomarkers have prognostic significance in people without diabetes or kidney disease is less studied. Using the Atherosclerosis Risk in Communities study, we followed individuals without diabetes or kidney disease for an average of 15 years after biomarker measurement to see if these biomarkers were associated with the development of kidney disease. We found that elevated levels of KIM-1, suPAR, TNFR-1, and TNFR-2 were associated with the development of kidney disease. These biomarkers may help identify individuals who would benefit from interventions to prevent the development of kidney disease.

Cartilaginous glycoprotein‑39 as laboratory marker of dynamics of course and effectiveness of treatment in patients with osteoarthritis

Backgrounds. Osteoarthritis (OA) occupies the first rank among diseases of the musculoskeletal system and accounts for 18.4% of the structure of this pathology. Currently, OA is considered as a disease in which there is a violation of the processes of remodeling (degradation and synthesis) of cartilage tissues, subchondral bone, joint capsule, tendon-ligamentous and muscular apparatus. An increased concentration of cartilage glycoprotein-39 (CGP-39) in the circulation is associated with inflammatory diseases and the processes of active tissue restructuring. To date, CGP-39 is a marker of chondrocyte activation and a sign of progressive OA, which determines its clinical significance. Along with this, the role of this biomarker in evaluating the effectiveness of anti-inflammatory therapy is relevant.The purpose. Comparative assessment of the quantitative content of CGP-39 in patients with knee joint OA, depending on the severity of inflammatory changes in the joints, the dynamics of the disease and the effectiveness of therapy.Methods. The study included 36 patients with OA and 30 practically healthy volunteers, identical in gender and age to the examined patients. The mean age of patients was 64 years, the average duration of the disease was 6 years. The diagnosis was determined according to the diagnostic criteria of the American College of Rheumatology (ACR, 1991). Along with the generally accepted clinical, laboratory and instrumental diagnosis of OA, in all patients and in control persons the levels of circulating CGP-39 were measured by enzyme immunoassay (ELISA). All examined patients received Ambene® Bio for 20 days, followed by repeated determination of the level of CGP-39. For statistical processing of the obtained data, we have used the programs Microsoft Excel 2007 and Statistica 10.0. Using the application program KRelRisk 1.1, the relative risk indicator for the studied factor was determined.Results. In the blood of patients with OA, a significant increase in the levels of CGP-39 has been found in comparison with the group of healthy individuals, which indicates increased degradation of articular cartilage in OA. As the severity of knee OA increased, the concentration of CGP-39 in the blood serum significantly raised, including in comparison with the control group. Reliable direct correlations were found between CGP-39 and clinical indices of knee OA severity. The content of CGP-39 in serum in patients with stage III radiological OA was significantly higher than in patients with stage I–II. The highest level of CGP-39 in patients with OA is associated with the presence of synovitis. TThe level of CGP-39 significantly decreased in patients with knee OA after a course of Ambene® Bio therapy, which confirms the participation of CGP-39 in inflammation on the one hand and the possibility of its use as an indicator of the effectiveness of anti-inflammatory therapy on the other hand.Conclusions. CGP-39 as a marker of inflammation and degradation of articular cartilage reflects the severity of the course of OA. The investigation of this biomarker is useful not only for diagnostic purposes, but also to assess the response to anti-inflammatory treatment in patients with knee OA.

Clam challenges high concentrations of algae diet environment: Growth performance, metabolism physiology, and related gene expression characteristics

Increasing coastal pollution leads to frequent algal blooms, but little is known on the effect of growth and development in organisms with increasing algal concentration. In recent years, the clam Cyclina sinensis has been an important marine economic shellfish that is widely cultivated. This study investigated the effects of different Chlorella concentrations (5 × 105 cell/L (Diet T1), 1.25 × 106 cell/L (Diet T2), 1.3 × 107 cell/L (Diet T3), 1.25 × 108 cell/L (Diet T4), and 1.5 × 109 cell/L (Diet T5)) on the growth, enzyme activities of digestive, immune, and respiratory metabolism, and hepatopancreatic transcriptome gene expression characteristic response of C. sinensis. Results showed that the shell length growth rate, shell height growth rate, and survival rate of C. sinensis significantly decreased as the Chlorella concentration reached 5 × 105 cell/L. The condition factor of C. sinensis in Diet T1 and the hepatopancreas index of C. sinensis in Diet T4 were significantly higher than those in the other groups (P < 0.05). The enzyme activities of superoxide dismutase increased significantly with the rise in concentration of Chlorella, but the highest lysozyme in the hepatopancreas of C. sinensis was observed in the Diet T1 group. The enzyme activities of acid phosphatase and amylase in Diet T3, trypsin in Diet T5, lipase and succinate dehydrogenase in Diet T4, and lactate dehydrogenase in Diet T1 of C. sinensis were significantly higher than those in the other groups (P < 0.05). These results suggest that some enzyme activities related to digestion, metabolism, and respiration are inhibited with the Chlorella concentration increased. Furthermore, transcriptome results showed that the expression levels of G6P, IGFBP, CHI3L1, FAS, MyHC, and other genes related to glucose, protein, and fatty acid metabolism of C. sinensis were significantly downregulated in the Diet T3 and Diet T5 groups. All these results suggest that Chlorella concentrations above 5 × 105 cell/L should be avoided in practice. This study provides a scientific basis for C. sinensis culture and further extends the understanding of the metabolism of bivalves with diet concentration.

Genetic Markers of Insulin Resistance and Atherosclerosis in Type 2 Diabetes Mellitus Patients with Coronary Artery Disease.

Type 2 diabetes mellitus (T2DM) is characterized by impaired insulin secretion on a background of insulin resistance (IR). IR and T2DM are associated with atherosclerotic coronary artery disease (CAD). The mechanisms of IR and atherosclerosis are known to share similar genetic and environmental roots. Endothelial dysfunction (ED) detected at the earliest stages of IR might be the origin of atherosclerosis progression. ED influences the secretion of pro-inflammatory cytokines and their encoding genes. The genes and their single nucleotide polymorphisms (SNPs) act as potential genetic markers of IR and atherosclerosis. This review focuses on the link between IR, T2DM, atherosclerosis, CAD, and the potential genetic markers CHI3L1, CD36, LEPR, RETN, IL-18, RBP-4, and RARRES2 genes.

OR30-5 Androgen-Regulated Secreted Factors Promote Tumorigenesis and Immune Suppression in Breast Cancer

Abstract Background Breast cancer (BC) is the most diagnosed cancer and the second-leading cause of cancer-associated death in women in the United States. BCs that express the estrogen and/or progesterone receptor (ER/PR) are treated with ER-targeting endocrine therapies that effectively reduce disease burden and extend lifespan. However, up to 50% of patients with ER+BCs will experience recurrence, often at a metastatic site, within 10 years of diagnosis. Novel targeted therapies are needed to treat both metastatic ER+BC resistant to current treatment strategies and chemoresistant ER-BCs.We identified chitinase 3–like-1 (CHI3L1/BRP-39/YKL-40) and prolactin inducible protein (PIP/GCDFP-15) as androgen receptor (AR)-regulated in BC. PIP is reported to be detectable in &amp;gt;90% of BCs, but its function in BC is not well-characterized. CHI3L1 is a glycosyl hydrolase family 18 protein member with reported roles in aging and numerous pathologies. During mammary gland involution, CHI3L1 contributes to a pro-tumorigenic wound-healing-like environment and increases macrophage infiltration. Further, CHI3L1 correlates with tumor-suppressive M2 macrophage infiltration in ER-BC. We previously reported that ER+ cells and patient-derived xenografts with activating mutations in the ESR1 gene encoding ER—a known mechanism of endocrine therapy resistance—had elevated AR and CHI3L1 gene and protein expression. Based on the above and our finding that metastatic BCs biopsies harboring ESR1 mutations exhibited increased AR expression, we hypothesized that AR regulation of CHI3L1 and PIP promote progression to metastatic disease in part through immune suppressive effects. Methods A ConTra(V3) transcription factor binding site analysis was performed on CHI3L1 and PIP promoters. CHI3L1 and PIP were measured via immunohistochemistry in xenograft tumors in mice treated with or without the androgen dihydrotestosterone (DHT). Immunoblotting, immunohistochemistry, and immunofluorescence approaches were also utilized to assess CHI3L1 protein expression in DHT-treated, tamoxifen resistant (TAMR), and long term E2 deprived (LTED) ER+BC cells. CIBERSORT analysis was conducted on the Firehouse Legacy TCGA dataset. Macrophage efferocytosis assays were performed on an IncuCyte Live Cell Imaging system. Results We identified highly conserved AR binding sites in the CHI3L1 and PIP promoters. CHI3L1 protein was elevated in TAMR and LTED BC cells, which was further elevated in BC cells with ESR1 mutations, as well as DHT-treated cells. Further, CHI3L1 and PIP were increased in xenograft tumors after DHT treatment. CIBERSORT analysis revealed that CHI3L1 and PIP high tumors had altered immune profiles, including differential macrophage levels. Finally, CHI3L1 and PIP treatment significantly reduced macrophage efferocytosis of BC cells (p ≤ 0.0001). Conclusions Our work suggests that AR-mediated regulation of CHI3L1 and PIP may alter immune cell infiltration/function in BC and may be important in endocrine therapy resistant ER+BCs. Further characterization and manipulation of these AR-regulated proteins is underway to test the potential to target and reverse their immune suppressive functions. Presentation: Tuesday, June 14, 2022 10:45 a.m. - 11:00 a.m.

GAAGs, COMP, and YKL-40 as Potential Markers of Cartilage Turnover in Blood of Children with Juvenile Idiopathic Arthritis Treated with Etanercept—Relationship with ADAMTS4, ADAMTS5, and PDGF-BB

We quantified galactosaminoglycans (GAAGs), oligomeric cartilage matrix protein (COMP), and human cartilage glycoprotein 39 (YKL-40) in blood obtained from juvenile idiopathic arthritis (JIA) before and during 2-year treatment with etanercept (ETA), as potential biomarkers of cartilage extracellular matrix (ECM) dysfunction and indicators of efficacy of biologic therapy. We also evaluated the relationship of the mentioned markers with the factors that regulate their metabolism, disintegrin and thrombospondin motif metalloproteinases 4 (ADAMTS4), ADAMTS5, and platelet-derived growth factor BB (PDGF-BB). Methods: We studied 38 children diagnosed with JIA and 45 healthy children. We quantified GAAGs by assessing the concentration of unsaturated disaccharide units formed by digestion of isolated glycosaminoglycans with chondroitinase ABC, while COMP, YKL-40, and PDGF-BB were quantified using immunoenzymatic methods. Results: Compared to the control group, GAAGs and COMP levels were significantly lower, while YKL-40 levels were higher in the blood of patients with aggressive JIA, qualified for ETA treatment. ETA therapy leading to clinical improvement simultaneously promoted normalization of COMP and YKL-40 levels, but not GAAGs. After 24 months of taking ETA, glycan levels were still significantly lower, relative to controls. GAAGs, COMP, and YKL-40 levels were significantly influenced by ADAMTS4, ADAMTS5, and PDGF-BB levels both before and during ETA treatment. Conclusions: The dynamics of changes in marker concentrations during treatment seem to indicate that measurement of COMP and YKL-40 levels can be used to assess the chondroprotective biological efficacy of therapy. In contrast, changes in GAAGs concentrations reflect systemic extracellular matrix transformations in the course of JIA.

Comparison of Short-Term Restorative Effects and Periodontal Health Status of Restorations Made of Different Materials in Full-Crown Restoration of Mandibular Premolar Tooth Defects.

Purpose To compare the short-term restorative effect and periodontal health status of restorations with different materials in full-crown restoration of mandibular premolar tooth defects. Methods A total of 105 cases (123 affected teeth) of mandibular premolar tooth defects who visited the Department of Stomatology between January 2019 and January 2020 were selected, of which 58 cases (68 affected teeth) restored with cobalt-chromium alloy porcelain crowns were included in the metal-ceramic crown (MCC) group and 47 cases (55 affected teeth) repaired by zirconium dioxide all-ceramic crowns were assigned to the zirconia crown (ZC) group. The restorative effect, inflammatory factors (high-sensitivity C-reactive protein (hs-CRP) and tumor necrosis factor-α (TNF-α)) and periodontal health indicators (human cartilage glycoprotein-39 (YKL-40), resistin, aspartate aminotransferase (AST), and alkaline phosphatase (ALP)) after treatment, and the occurrence of adverse reactions were compared between the two groups. Results The ZC group outperformed the MCC group in terms of margin fitness, restoration fracture, gingival condition, and color matching. After restoration, hs-CRP and TNF-α were statistically decreased in both groups, while YKL-40, resistin, AST, and ALP were significantly increased; and compared with the MCC group, hs-CRP, TNF-α, YKL-40, resistin, AST, and ALP were lower in the ZC group. The ZC group also scored statistically higher in retention effect, aesthetic effect, masticatory function, voice function, and comfort. Moreover, the ZC group had a higher total effective rate and a lower incidence of adverse reactions than the MCC group, with statistical significance. Conclusions Zirconia dioxide all-ceramic crowns contribute to a better short-term restorative effect in the full-crown restoration of mandibular premolar tooth defects, with little impact on periodontal health and high patient satisfaction, which deserves popularization and application.

Effects of Sacubitril/Valsartan on biomarkers of fibrosis and inflammation in patients with heart failure with reduced ejection fraction

AimsTo evaluate the circulating levels of remodeling biomarkers procollagen type 1 C-terminal propeptide (PICP), human cartilage glycoprotein-39 (YKL-40), plasma renin activity (PRA), aldosterone (Aldo) as well as clinical and echocardiographic parameters in patients with heart failure with reduced ejection fraction (HFrEF), before and after treatment with Sacubitril/Valsartan (S/V).Methods and resultsA total of 26 consecutive patients with HFrEF on stable clinical conditions were studied. Clinical, echocardiographic parameters and circulating biomarkers were measured at baseline, after 30 and 60 days of S/V treatment. Both systolic blood pressure (SBP) and diastolic blood pressure (DBP) decreased, from 126 ± 15 to 113 ± 4 mmHg (p < 0.001) and from 77 ± 11 to 72 ± 9 mmHg (p = 0.005), respectively, at the end of study. Concomitantly, left ventricular ejection fraction (LVEF) increased by 22.8% from 29.5 ± 5% to 36.2 ± 5%, (p < 0.001) and indexed left ventricular end-systolic volume (LVESVi) decreased by 12% from 38.6 ± 8.7 ml/m2 to 34.0 ± 10.0 ml/m2. (p = 0.007). Circulating levels of PICP, YKL-40, PRA and Aldo decreased by − 42.2%, − 46.8%, − 79.1% and − 76.7%, respectively (p < 0.001 for all), the decrements being already maximal within 30 days of S/V treatment. No significant changes of plasma electrolytes and creatinine were observed during the study (all p > 0.05).ConclusionsA decrease of circulating markers of inflammation and fibrosis during chronic treatment with S/V is associated with an improvement of hemodynamic and echographic parameters in patients with HRrEF. These data are compatible with an anti-fibrotic and anti-inflammatory effect of S/V, that may contribute to the beneficial outcomes of the drug in this clinical setting.

Role of human cartilage glycoprotein-39 and vitamin C in the development and clinical course of knee arthrosis in combination with coronary artery disease, hypertension, dyslipidemia, cholelithiasis

Aim. To study the role of human cartilage glycoprotein-39 (HC gp-39) and vitamin C in the development and clinical course of knee arthrosis (gonarthrosis) in combination with coronary artery disease (CAD), hypertension (HTN), dyslipidemia, cholelithiasis, depending on inflammatory changes in the joints, X-ray stage, pain intensity and the degree of joints’ functional impairment.Material and methods. We examined 140 patients with gonarthrosis in combination with CAD, HTN, dyslipidemia, cholelithiasis, who were treated in the rheumatology department of the North Ossetian State Medical Academy. In all patients, the main clinical parameters of gonarthrosis were assessed, and the content of HC gp-39, vitamin C, C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and serum lipids was determined.Results. An increase in HC gp-39 levels was revealed in all examined patients. The highest rate of HC gp-39 was found in patients with X-ray stage III arthrosis, especially with reactive synovitis. A decrease in blood serum concentration of vitamin C was revealed depending on gonarthrosis stage. A significant correlation was noted between HC gp-39 level and main clinical indices of gonarthrosis, as well as CRP, ESR, cholesterol, atherogenic coefficient, and vitamin C.Conclusion. Changes in the levels of HC gp-39, vitamin C, CRP and lipid profile reflect the inflammation severity, which is an important component of the pathogenesis of osteoarthritis, atherosclerosis, and cholelithiasis.

The role of YKL-40 in the pathogenesis of autoimmune diseases: a comprehensive review.

YKL-40, a chitinase-3-like protein 1 (CHI3L1) or human cartilage glycoprotein 39 (HC gp-39), is expressed and secreted by various cell-types including macrophages, chondrocytes, fibroblast-like synovial cells and vascular smooth muscle cells. Its biological function is not well elucidated, but it is speculated to have some connection with inflammatory reactions and autoimmune diseases. Although having important biological roles in autoimmunity, there were only attempts to elucidate relationships of YKL-40 with a single or couple of diseases in the literature. Therefore, in order to analyze the relationship between YKL-40 and the overall diseases, we reviewed 51 articles that discussed the association of YKL-40 with rheumatoid arthritis, psoriasis, systemic lupus erythematosus, Behçet disease and inflammatory bowel disease. Several studies showed that YKL-40 could be assumed as a marker for disease diagnosis, prognosis, disease activity and severity. It is also shown to be involved in response to disease treatment. However, other studies showed controversial results particularly in the case of Behçet disease activity. Therefore, further studies are needed to elucidate the exact role of YKL-40 in autoimmunity and to investigate its potential in therapeutics.

Role of GDF-15, YKL-40 and MMP 9 in patients with end-stage kidney disease: focus on sex-specific associations with vascular outcomes and all-cause mortality

BackgroundSex differences are underappreciated in the current understanding of cardiovascular disease (CVD) in association with chronic kidney disease (CKD). A hallmark of CKD is vascular aging that is characterised, amongst others, by; systemic inflammation, microbiota disbalance, oxidative stress, and vascular calcification—features linked to atherosclerosis/arteriosclerosis development. Thus, it is the necessary to introduce novel biomarkers related to athero-/arteriosclerotic damage for better assessment of vascular ageing in patients CKD. However, little is known about the relationship between uraemia and novel CVD biomarkers, such as growth differentiation factor-15 (GDF-15), cartilage glycoprotein-39 (YKL-40) and matrix metalloproteinase-9 (MMP-9). Therefore, we hypothesise that there are sex-specific relationships between GDF-15, YKL-40, MMP-9 levels in end-stage kidney disease (ESKD) patients in relation to gut microbiota, vascular calcification, inflammation, comorbidities, and all-cause mortality.MethodsESKD patients, males (n = 151) and females (n = 79), not receiving renal replacement therapy were selected from two ongoing prospective ESKD cohorts. GDF-15, YKL-40 and MMP9 were analysed using enzyme-linked immunosorbent assay kits. Biomarker levels were analysed in the context of gut microbiota-derived trimethylamine N-oxide (TMAO), vascular calcification, inflammatory response, oxidative stress, comorbidities, and all-cause mortality.ResultsIncreased GDF-15 correlated with higher TMAO in females only, and with higher coronary artery calcification and IL-6. In females, diabetes was associated with elevated GDF-15 and MMP-9, whilst males with diabetes only had elevated GDF-15. No associations were found between biomarkers and CVD comorbidity. Deceased males and females had higher GDF-15 concentrations (p = 0.01 and p < 0.001, respectively), meanwhile only YKL-40 was increased in deceased males (p = 0.02).ConclusionsIn conclusion, in males GDF-15 and YKL-40 were related to vascular calcification, inflammation, and oxidative stress, whilst in females GDF-15 was related to TMAO. Increased levels of YKL-40 and GDF-15 in males, and only GDF-15 in females, were associated with all-cause mortality. Our findings suggest that sex-specific associations of novel CVD biomarkers have a potential to affect development of cardiovascular complications in patients with ESKD.

MO009ROLE OF GDF-15, YKL-40, AND MMP-9 IN PATIENTS WITH ESKD: FOCUS ON SEX SPECIFIC ASSOCIATIONS WITH VASCULAR OUTCOMES AND ALL-CAUSE MORTALITY

Abstract Background and Aims This study was performed on behalf of the GOING-FWD Consortium. The importance of sex difference in the development of cardiovascular complications is well appreciated, however, further studies are warranted to assess if sex specific pathophysiological changes occur under uremic environment. Little is known about the relationship between the uremic phenotype and novel cardiovascular biomarkers like growth differentiation factor 15 (GDF-15), cartilage glycoprotein 39 (YKL-40), and matrix metalloproteinase 9 (MMP9), and if they have a sex specific effects in relation to inflammation, vascular remodelling, cardiovascular outcomes and all-cause mortality. Therefore, we hypothesise that there is a sex specific relationship between GDF-15, YKL-40, MMP9 and vascular outcomes defined as athero-/arteriosclerosis and vascular calcification. Method ESKD patients (n = 231), males (n = 152) and females (n = 79), not receiving renal replacement therapy were selected from two ongoing prospective CKD cohorts from the Division of Renal Medicine, Karolinska University Hospital, Sweden. Three putative CVD biomarkers, GDF-15, YKL-40 and MMP9 were analysed using enzyme-linked immunosorbent assay (ELISA) kits. Biomarker level/activity was analysed in the context of gut microbiota derived TMAO, vascular calcification (determined as CAC score on CT scans, aortic valve calcification (AVC) and medial calcification on epigastric artery biopsies), inflammatory response, oxidative stress and all-cause mortality. Results The levels of investigated biomarkers have not differed between female and male patients with ESKD. However, GDF-15 correlated with TMAO in females from more severe cohort that are not receiving renal replacement therapy yet, whereas in males increased GDF-15 level was associated with higher Agatston score on CT-scans, inflammatory biomarker (IL-6) and oxidative stress biomarker 8-OHdG. None of biomarkers was related to intimal medial calcification assessed in epigastric artery biopsies. Comorbidity analysis revealed elevated GDF-15 levels in both females and males, while increased MMP-9 levels were observed only in females with diabetes mellitus, but not with CVD. When assessing death of any cause in our cohorts, both deceased males and females had higher GDF-15 concentration (p = 0.01 and p &amp;lt; 0.001, respectively), meanwhile only YKL-40 level was increased in deceased males (p = 0.02). Conclusion In summary, here we report that in males GDF-15 and YKL-40 were related to vascular calcification, inflammageing and oxidative stress, while in females the relationship of GDF-15 with TMAO was observed. Higher MMP-9 level was seen in males without diabetes and in females with confirmed diabetes mellitus. YKL-40 increase in males and GDF-15 in both males and females were associated with all-cause mortality. Our findings suggest that sex specific associations exist and those could have a potential to affect development of cardiovascular complications in patients with ESKD.