Clinical Utility of Plasma Glypican-3 and Osteopontin as Biomarkers of Hepatocellular Carcinoma

BACKGROUND/AIMS: alpha-Fetoprotein (AFP) is the biomarker most widely used to detect hepatocellular carcinoma (HCC), despite its suboptimal diagnostic accuracy. Glypican-3 (GPC3) and osteopontin (OPN) are secreted glycoproteins that are reportedly associated with tumorigenesis and metastasis. This study was conducted to evaluate the clinical utility of using plasma GPC3 and OPN as diagnostic biomarkers for HCC. METHODS: We measured the plasma levels of GPC3 and OPN in 120 HCC and 40 chronic liver disease (CLD) patients via an enzyme-linked immunosorbent assay. The diagnostic accuracy of each tumor marker was evaluated using receiver operating characteristic (ROC) curve analysis. RESULTS: The GPC3 levels in the HCC patients (75.8 ng/mL) were significantly higher (p=0.020) than the levels in patients with CLD (66.4 ng/mL). The area under the ROC curve (AUROC) values for GPC3 and OPN were 0.62 and 0.51, respectively. In subgroup analyses, including subgroups of HCC patients with low serum AFP and PIVKA II levels, the AUROC of GPC3 remained relatively high (0.66), and GPC3 showed a high sensitivity (62.1%) for detecting small HCC tumors. CONCLUSIONS: The plasma levels of GPC3 and OPN demonstrated low diagnostic accuracy for HCC. However, GPC3 may have a complementary role in diagnosing HCC in patients with nondiagnostic levels of conventional tumor markers and with small-sized tumors.

High Serum Osteopontin Levels Are Associated with Low Bone Mineral Density in Postmenopausal Women

Osteopontin (OPN) is an acidic, noncollagenous matrix protein produced by the bone and kidneys. It is reportedly involved in bone resorption and formation. We examined the association between serum OPN levels and bone mineral density in postmenopausal women. Premenopausal women (n=32) and postmenopausal women (n=409) participated in the study. We measured serum osteopontin levels and their relationships with bone mineral density and previous total fragility fractures. The postmenopausal women had higher mean serum OPN levels compared to the premenopausal women (43.6+/-25.9 vs 26.3+/-18.6 ng/mL; P<0.001). In the postmenopausal women, high serum OPN levels were negatively correlated with mean lumbar bone mineral density (BMD) (r=-0.113, P=0.023). In a stepwise multiple linear regression model, serum OPN levels were associated with BMD of the spine, femoral neck, and total hip after adjustment for age, body mass index, smoking, and physical activity in postmenopausal women. However, serum OPN levels did not differ between postmenopausal women with and without fractures. Postmenopausal women exhibit higher serum OPN levels than premenopausal women and higher serum OPN levels were associated with low BMD in postmenopausal women.

Osteopontin: A Novel Protein Molecule.

Osteopontin (OPN) is phosphorylated sialic acid –rich non-collagenous bone matrix protein. OPN is found in several biological fluids including human plasma, serum, breast milk and urine.OPN was named for its function as a bridge between cells and minerals. OPN has been implicated as an important factor in bone remodeling. Osteopontin is expressed in immune cells, including macrophages, neutrophils, dendritic cells with varying kinetics. OPN influences cell mediated immunity and has Th 1 cytokine functions .OPN is overexpressed in cancers of lung, breast, colorectal, stomach and ovary.OPN is found in atheromatous plaques within arteries. OPN plays an important role during both acute and chronic inflammation. OPN is upregulated in tissues during several pathological process including atherosclerosis, valve stenosis, myocardial infarction and rheumatic arthritis. OPN is a key cytokine regulating tissue repair. OPN’s plasma levels are elevated in overweight and obesity.

Plasma and synovial fluid osteopontin levels in patients with knee osteoarthritis: relation to radiological grade

To investigate osteopontin [OPN] levels in both plasma and synovial fluid of patients with primary knee osteoarthritis [OA] and their relationship with radiological grade. Sixty patients had knee OA and 30 control subjects were included. Anteroposterior knee radiographs were taken to determine the disease severity of the affected knee. The radiographic grading of OA in the knee was performed by using the Kellgren-Lawrence grading. Osteopontin levels in the plasma and synovial fluid were measured using enzyme-linked immunosorbent assay and compared. OA patients had higher plasma osteopontin concentrations compared to healthy controls [p < 0.000]. Osteopontin levels in synovial fluid were significantly higher with respect to plasma sample [r = 0.694, p < 0.000]. The mean plasma levels of osteopontin in KL grade 4 were greater than those in KL grade 3, and the difference was statistically significant [p < 0.01]. The plasma osteopontin levels significantly correlated with the severity of disease [r = 0.870, p < 0.000]. The synovial fluid levels of osteopontin also correlated with disease severity as regarding the radiological grade [r = 0.817, p < 0.000]. Osteopontin in plasma and synovial fluid is related to progressive joint damage in knee OA. Osteopontin may serve as a biochemical marker for determining disease severity as regarding radiological grade

Osteopontin expression and its possible functions in the aortic disorders and coronary artery disease / A expressão da osteopontina e as suas funções possíveis nas desordens aórticas e doença arterial coronariana

BACKGROUND: Osteopontin (OPN) has been verified to be closely associated with oncogenesis and remodeling processes. But this cytokine was rarely assessed in the presence of aortopathies, especially acute aortic dissection. The aim of the present study was to evaluate the expressions of OPN by way of molecular biological approaches so as to offer a better understanding of the possible mechanisms of the aortopathies. METHODS: Consecutive patients with type A acute aortic dissection (20 patients), aortic aneurysm (nine patients) or coronary artery disease (21 patients) referred to this hospital for surgical operations were enrolled into this study. Blood samples of the surgical patients after systematic heparinization, and control fast morning blood samples drawn from 21 young healthy volunteers who had no evidence of any healthy problems were investigated for enzyme linked immunosorbent assay (ELISA). The surgical specimens of the aortic tissues collected from the surgical patients during the operations were obtained for quantitative realtime reverse transcription polymerase chain reaction (RT-PCR) for OPN mRNA, western blot assay for OPN protein, and for immunohistochemical staining of OPN. Ascending aortic tissues from the autopsies of the healthy individuals dying of accident were obtained as controls of immunohistochemistry. RESULTS: By quantitative RT-PCR, the expressions of OPN mRNA were all upregulated in all three surgical groups. The quantitative results did not reveal any intergroup differences. Western blot assay revealed that OPN was positive with similar intensities of expressions in all three surgical groups. Quantitative western blot analyses of OPN expressions did not show any significance between groups. The OPN expressions by ELISA in the aortic tissue were 3.09311 ± 1.65737, 3.40414 ± 1.15095, and 1.68243 ± 0.31119 pg/mg protein in the aortic dissection, aortic aneurysm, and coronary artery disease groups, respectively. The OPN level of the patients with coronary artery disease was much lower than those with aortic dissection (P = 0.033) or with aortic aneurysm (P = 0.019). By unparametric tests, there were significant differences in the aortic OPN contents among aortic dissection, aortic aneurysm and coronary artery disease groups (P < 0.01). A significant direct correlation was present between plasma OPN concentration and the time interval from the onset to surgery of aortic dissection (Y = 0.1420X + 2.4838, r² = 0.5623, r = 0.750, P = 0.032). By immunohistochemistry, OPN was expressed in the aortic cells: in the intima, it was weaker in all three surgical groups in comparison with the healthy control; in the media, it was weak in the aortic dissection, intense positive in aortic aneurysm, focal positive in the coronary artery disease, but evenly positive in the healthy control groups; and in the adventitia, it was positive in the aortic dissection, coronary artery disease and healthy control groups, but weak positive in the aortic aneurysm group. CONCLUSION: These data may provide evidences that OPN may play a role in the pathogenesis of aortopathies including aortic dissection, aortic aneurysm, and coronary artery disease. OPN might be of potential perspective as a clinically diagnostic tool in the evaluations of the complex remodeling process incorporating vascular injury and repair.

OBJETIVOS: A osteopontina (OPN) está estreitamente associada com os processos de oncogênese e remodelação. Entretanto, essa citocina era raramente avaliada na presença de aortopatias, especialmente na dissecção aórtica aguda. O objetivo do presente estudo foi avaliar a expressão de OPN por meio de abordagens moleculares biológicas, de modo a oferecer uma melhor compreensão dos possíveis mecanismos das aortopatias. MÉTODOS: Pacientes consecutivos com um tipo de dissecção aguda da aorta (20 pacientes), aneurisma da aorta (nove pacientes) ou doença arterial coronária (21 pacientes) foram incluídos neste estudo. As amostras de sangue depois da heparinização sistemática e de 21 voluntários jovens e saudáveis não apontaram nenhuma evidência de qualquer problema ao serem investigados por ensaio imunoenzimático (ELISA). Os espécimes cirúrgicos dos tecidos aórtica coletados dos pacientes durante as operações foram obtidos para a reação de transcrição reversa quantitativa em tempo real em cadeia da polimerase (RT-PCR) para OPN mRNA, técnica de Western blot para a proteína OPN, e imunohistoquímica de OPN. Amostras da aorta de indivíduos saudáveis que morreram de acidente foram obtidos para controle imunohistoquímico. RESULTADOS: Com uso do RT-PCR quantitativo, as expressões de OPN mRNA foram suprarreguladas em todos os três grupos cirúrgicos. Os resultados quantitativos não revelaram quaisquer diferenças intergrupais. Western blot revelou que OPN foi positiva com intensidade semelhante de expressões em todos os três grupos. As análises quantitativas Western blot de expressões OPN não apresentaram significâncias entre os grupos. As expressões OPN medidas pelo teste ELISA no tecido aórtico foram 3,09311 ± 1,65737, 3,40414 ± 1,15095 e 1,68243 ± 0,31119 pg/mg de proteína na dissecção de aorta, aneurisma da aorta, e grupos de doença arterial coronariana, respectivamente. O nível de OPN dos pacientes com doença arterial coronariana foi muito menor do que aqueles com dissecção aórtica (P = 0,033) ou com aneurisma da aorta (P = 0,019). Testes não-paramétricos apontaram diferenças significativas nos teores de aorta OPN entre dissecção aórtica, aneurisma da aorta e grupos com doença arterial coronariana (P <0,01). Uma correlação direta significativa estava presente entre a concentração plasmática OPN e o intervalo de tempo entre o início da cirurgia de dissecção de aorta (Y = 2,4838 + 0,1420X, r² = 0,5623, r = 0,750, P = 0,032). Pela imunohistoquímica, a OPN foi expressa em células aórticas: na íntima, foi fraca em todos os três grupos cirúrgicos em comparação ao grupo saudável; na média, era fraca na dissecção aórtica, positiva intensa no aneurisma de aorta, focal positivo na doença arterial coronariana, mas igualmente positiva no grupo controle; e na adventícia, positiva para a dissecção da aorta, doença arterial coronariana e grupos de controle saudáveis, mas fraca positiva no grupo de aneurisma da aorta. CONCLUSÃO: Estes dados fornecem evidências de que a OPN pode desempenhar um papel na patogênese da aortopatias, incluindo dissecção aórtica, aneurisma da aorta R e doença arterial coronariana. OPN tem perspectiva potencial como ferramenta de diagnóstico clínico nas avaliações do processo de remodelação complexa, incluindo lesão vascular e de reparação.

Prognostic value of serum osteopontin in hepatocellular carcinoma patients treated with transarterial chemoembolization / 대한간학회지

BACKGROUND/AIMS: Osteopontin (OPN) is overexpressed in hepatocellular carcinoma (HCC) with postoperative recurrence or extrahepatic metastasis. However, its prognostic value in patients treated with transarterial chemoembolization (TACE) is unclear. We investigated the utility of serum OPN levels and changes therein as prognostic markers in HCC patients who have received TACE. METHODS: Forty-six patients with HCC were enrolled. Serum OPN levels were measured before and 4 weeks after TACE. Serum biochemistry and computed tomography (CT) scans were analyzed. We evaluated baseline serum OPN levels and subsequent changes therein in relation to tumor responses and cumulative survival rates following TACE. A decreasing pattern was defined as a decrease after TACE of more than 10% relative to baseline levels. A "responder" was defined as a patient who exhibited a tumor necrosis rate of higher than 50% on the follow-up CT scan. RESULTS: Higher initial serum OPN levels were associated with a large tumor, portal vein invasion, and an advanced tumor stage. Patients who had lower initial serum OPN levels and those who exhibited decreasing patterns after TACE tended to have more favorable tumor responses (P=0.043 and 0.055, respectively) and exhibited better cumulative survival rates (P=0.036 and 0.030, respectively). However, the initial serum OPN level and subsequent changes in serum OPN levels were not independent predictors for survival on multivariate analysis. CONCLUSIONS: Serum OPN levels were significantly higher in patients with advanced HCC. In addition, HCC patients with low pretreatment serum OPN levels and those for whom serum OPN declined following TACE exhibited better tumor responses and survived for longer.

Comparative estimation of serum osteopontin tumor necrosis factor-alpha and IL-6 as an indictor for immune dysregulation in nasopharyngeal carcinoma

The present study was designed to evaluate serum levels of osteopontin [OPN], tumor necrosis factor-alpha [TNF-alpha] and interleukin-[IL]-6 in pre-treatment samples obtained from patients with biopsy confirmed nasopharyngeal carcinoma [NPC]. The study included 28 NPC patients; 20 males and 8 females with mean age of 56.8 +/- 8 years. Cervical lymphadenopathy was the main presenting symptom in 19 patients [67.9%], recurrent unexplained attacks of epistaxis in 16 patients [57.1%] and 7 patients [25%] had secretory otitis media. Patients were clinically categorized using TNM staging and underwent nasopharyngoscopy and biopsy taking for pathological examination and grading according to the World Health Organization [WHO] types. All patients received chemo-radiotherapy and completed their follow-up at ENT outpatient clinic. Pre-and post-treatment blood samples were collected for estimation of serum level of osteopontin [OPN], tumor necrosis factor-a [TNF-alpha] and interleukin-[lL]-6. Blood samples were obtained from 10 healthy volunteers as control group. Pretreatment serum levels of estimated parameters were significantly higher compared both to control levels and to post-treatment levels. However, despite treatment induced significant decrease of serum levels of estimated parameters, their levels still significantly higher compared to control levels. There was a positive significant correlation between TNM clinical staging and serum levels of OPN, TNF alpha and IL-6, Also, WHO pathological types showed a positive significant correlation with serum levels of OPN and lL-6, but the correlation with TNF-alpha was positive non-significant. Using ROC analysis for estimated parameters as screening test for WHO type I lesions defined estimation of serum OPN as a good screening test to detect early lesions and defined 2 cutoff points for serum OPN; namely: 265 and 298 ng/ml, had identical screening power however, cutoff point at 265 ng/ml showed significantly higher of sensitivity rate [89.3%]. NPC is associated with immune dysregulation in favor of Th1 side and elevated OPN pre-treatment serum levels that could be used as screening test for early cases of NPC as a preliminary screening test with cutoff point at 265 ng/ml as discriminative value

Plasma concentration of osteopontin [OPN] in children with systemic lupus erythematosus: relationship with disease activity

Osteopontin [OPN] is an important bone matrix mediator found to have key roles in inflammation and immunity. OPN is a cytokine which can play a number of roles in promoting activation of T lymphocyte, regulating balance between T-helper 1 and T-helper 2, participating in cell-induced immunologic response and stimulating B lymphocyte to express multi-clone antibodies. Overexpression of OPN has been associated with the development of the autoimmune/lymphoproliferative syndrome. The aim of our present study was to analyze the possible correlation between the plasma concentration of OPN and disease activity in children with Systemic Lupus Erythematosus [SLE]. We also investigated the correlation between plasma IL-18 and OPN concentrations to further confirm the association of OPN with disease activity. We measured the plasma concentration of OPN, and the plasma proinflammatory IL-18 concentration in 40 SLE patients with or without renal disease [RSLE group and SLE group, respectively] and in 30 sex-and age-matched controls using enzyme immunoassay. Plasma OPN concentrations were significantly higher in RSLE and SLE patients than in the controls [p=0.000 and p=0.002]. Increase in OPN concentration correlated positively and significantly with SLE disease activity index in all SLE patients [r=0.34; p=0.04]. In RSLE patients, plasma OPN concentration showed a significant positive correlation with proinflammatory cytokine IL-18 concentration [r=0.48; p=0.004]. In conclusion, The above results suggest that the production of OPN is associated with the inflammatory process and SLE development, and may serve as a potential disease marker of SLE

Clinical Significance of Serum Osteopontin in Patients with Multiple Myeloma / 대한진단검사의학회지

BACKGROUND: Angiogenesis and osteoclastogenesis are increased in the bone marrow of multiple myeloma (MM) patients in parallel with the tumor progression. Osteopontin (OPN) is a multifunctional protein that is involved in angiogenesis and bone destruction and, eventually, in tumor progression in MM. OPN is known to increase in MM patients as the disease progresses and bone is destroyed. We studied the clinical usefulness of OPN as a monitoring marker for treatment response in patients with MM. METHODS: We obtained 70 serial sera from 27 MM patients and 14 sera from healthy individuals. OPN was measured by a sandwich ELISA method. The hospital records were reviewed, and the clinically important markers for monitoring the treatment response, such as monoclonal component, immunoglobulin, free light chain, and hemoglobin, etc, were analyzed together with OPN levels. RESULTS: There was no significant difference in OPN levels between MM patients and healthy controls. OPN showed no significant correlations with the markers used for monitoring of treatment response such as M component, immunoglobulin, and free light chain levels. There was no difference in OPN levels between the 3 groups classified by the amount of M component. In addition, OPN levels showed no compatible changes to the treatment response of MM patients. CONCLUSIONS: Although OPN has been known to have an important role in the formation and progression of MM by involving angiogenesis and bone destruction, our results show that OPN is not valuable as a clinical marker for monitoring the treatment response in MM patients because of inconsistency in its levels in MM patients.

Plasma osteopontin concentration in lupus nephritis

To measure plasma concentration of osteopontin and to correlate these levels with clinical, laboratory, disease activity and histopathological parameters in SLE patients. This study was performed on 20 SLE patients and 10 normal control subjects. Plasma osteopontin concentrations were measured by ELISA technique for both patients and control groups. All patients underwent renal biopsies within 3 months of onset of proteinuria or hematuria. There was a highly significant difference between patients and control groups as regards plasma osteopontin concentration [p<0.001]. There was a significant positive correlation between osteopontin and SLEDAI [p<0.001] and activity index of renal biopsies [p<0.05]. Also there was a highly significant differences as regard osteopontin plasma concentration between patients with and without renal affection [p<0.001]. Osteopontin has been shown at least partly to account for SLE nephritis probably through predominance of Th[1]-type response in both peripheral and renal tissue. Further investigation of this mechanism in lupus nephritis may allow the design of new therapeutic strategies of lupus nephritis such as manipulation of Th[1]/Th[2] and down-regulation of Th[1]-response