Elashoff D, Zhou H, Reiss JK, Wang J, Henson B, Hu S, Arellano-Garcia M, Sinha UK, Le AD, Messadi D, Wang MB, Nabili V, Lingen MW, Morris D, Randolph TW, Feng Z, Akin D, Kastratovic DA, Chia D, Abemayor E, Wong DT. Pre-Validation of Salivary Biomarkers for Oral Cancer Detection. (2012) Cancer Epidemiol Biomarkers Prev : Show Abstract · Added February 5, 2012BACKGROUND: Oral cancer is the sixth most common cancer with a five-year survival rate of approximately 60%. Presently there are no scientifically credible early detection techniques beyond conventional clinical oral examination. The goal of this study is to validate if the 7 mRNAs and 3 proteins previously reported biomarkers are capable of discriminating patients with oral squamous cell carcinomas (OSCC) from healthy subjects in independent cohorts and by a National Cancer Institute (NCI)- Early Detection Research Network (EDRN) Biomarker Reference Laboratory (BRL).METHODS: 395 subjects from 5 independent cohorts based on case-controlled design were investigated by 2 independent laboratories, UCLA discovery laboratory and NCI-EDRN Biomarker Reference Laboratory (BRL). RESULTS: Expression of all 7 mRNA and 3 protein markers was increased in OSCC versus controls in all 5 cohorts. With respect to individual marker performance across the 5 cohorts, the increase in IL-8 and SAT were statistically significant and remained top performers across different cohorts in terms of sensitivity and specificity. A previously identified multiple marker model demonstrated an area under the receiver operating characteristic (ROC)-curve for prediction of OSCC status ranging from of 0.74 to 0.86 across the cohorts.CONCLUSIONS: The validation of these biomarkers demonstrated their feasibility in the discrimination of OSCC from healthy controls. Established assay technologies are robust enough to perform independently. Individual cutoff values for each of these markers and for the combined predictive model need to be further defined in large clinical studies. Impact: Salivary proteomic and transcriptomic biomarkers can discriminate oral cancer from control subjects. | Publication | 22301830 (PMID) 10.1158/1055-9965.EPI-11-1093 (DOI) |
Xiao H, Zhang L, Zhou H, Lee JM, Garon EB, Wong DT. Proteomic analysis of human saliva from lung cancer patients using two-dimensional difference gel electrophoresis and mass spectrometry. (2011) Mol Cell Proteomics : Show Abstract · Added November 19, 2011Lung cancer is often asymptomatic or causes only nonspecific symptoms in its early stages. Early detection represents one of the most promising approaches to reduce the growing lung cancer burden. Human saliva is an attractive diagnostic fluid because its collection is less invasive than that of tissue or blood. Profiling of proteins in saliva over the course of disease progression could reveal potential biomarkers indicative of oral or systematic diseases, which may be used extensively in future medical diagnostics. There were 72 subjects enrolled in this study for saliva sample collection according to the approved protocol. Two-dimensional difference gel electrophoresis (2D-DIGE) combined with MS was the platform for salivary proteome separation, quantification and identification from two pooled samples. Candidate proteomic biomarkers were verified and pre-validated by using immunoassay methods. There were 16 candidate protein biomarkers discovered by 2D-DIGE and MS. Three proteins were further verified in the discovery sample set, pre-validation sample set and lung cancer cell lines. The discriminatory power of these candidate biomarkers in lung cancer patients and healthy control subjects can reach 88.5% sensitivity and 92.3% specificity with AUC=0.90. This preliminary data report demonstrates that proteomic biomarkers are present in human saliva when people developed lung cancer. The discriminatory power of these candidate biomarkers indicating that a simple saliva test might be established for lung cancer clinical screening and detection. | Publication | 22096114 (PMID) 10.1074/mcp.M111.012112 (DOI) |
Lee YH, Kim JH, Zhou H, Kim BW, Wong DT. Salivary transcriptomic biomarkers for detection of ovarian cancer: for serous papillary adenocarcinoma. (2011) J Mol Med (Berl) : Show Abstract · Added November 19, 2011Ovarian cancer is the most lethal gynecological cancer due to lack of clear symptom and reliable screening biomarker in the early stage. The capability to detect the initiation of malignancy with a sensitive and effective approach is one of the most desirable goals for ovarian cancer therapy. In this study, we spearheaded noninvasive detection of ovarian cancer by salivary transcriptomic biomarkers, and evaluated the clinical utilities of discovered biomarkers using a clinical case-control study. To find salivary mRNA biomarkers, salivary transcriptomes in 11 ovarian cancer patients and 11 matched controls were profiled by Affymetrix HG-U133-Plus-2.0 array. The biomarker candidates selected from the microarray results were then subjected to clinical validation by RT-qPCR using an independent sample cohort including 21 ovarian cancer patients and 35 healthy controls. Seven downregulated mRNA biomarkers were validated. The logistic regression model revealed the combination of five validated biomarkers (AGPAT1, B2M, BASP2, IER3, and IL1B) can significantly discriminate ovarian cancer patients (n = 21) from the healthy controls (n = 35), yielding a receiver operating characteristic plot, area under the curve value of 0.909 with 85.7% sensitivity and 91.4% specificity. In summary, we have demonstrated that the RNA signatures in saliva could serve as biomarkers for detection of ovarian cancer with high sensitivity and specificity. This emerging approach with high-throughput, noninvasive, and effective advantages provides a feasible means for detection of systemic cancer, and opens a new avenue for early disease detection. | Publication | 22095100 (PMID) 10.1007/s00109-011-0829-0 (DOI) |
Farrell JJ, Zhang L, Zhou H, Chia D, Elashoff D, Akin D, Paster BJ, Joshipura K, Wong DT. Variations of oral microbiota are associated with pancreatic diseases including pancreatic cancer. (2011) Gut : Show Abstract · Added October 15, 2011ObjectiveThe associations between oral diseases and increased risk of pancreatic cancer have been reported in several prospective cohort studies. In this study, we measured variations of salivary microbiota and evaluated their potential associations with pancreatic cancer and chronic pancreatitis.MethodsThis study was divided into three phases: (1) microbial profiling using the Human Oral Microbe Identification Microarray to investigate salivary microbiota variation between 10 resectable patients with pancreatic cancer and 10 matched healthy controls, (2) identification and verification of bacterial candidates by real-time quantitative PCR (qPCR) and (3) validation of bacterial candidates by qPCR on an independent cohort of 28 resectable pancreatic cancer, 28 matched healthy control and 27 chronic pancreatitis samples.ResultsComprehensive comparison of the salivary microbiota between patients with pancreatic cancer and healthy control subjects revealed a significant variation of salivary microflora. Thirty-one bacterial species/clusters were increased in the saliva of patients with pancreatic cancer (n=10) in comparison to those of the healthy controls (n=10), whereas 25 bacterial species/clusters were decreased. Two out of six bacterial candidates (Neisseria elongata and Streptococcus mitis) were validated using the independent samples, showing significant variation (p<0.05, qPCR) between patients with pancreatic cancer and controls (n=56). Additionally, two bacteria (Granulicatella adiacens and S mitis) showed significant variation (p<0.05, qPCR) between chronic pancreatitis samples and controls (n=55). The combination of two bacterial biomarkers (N elongata and S mitis) yielded a receiver operating characteristic plot area under the curve value of 0.90 (95% CI 0.78 to 0.96, p<0.0001) with a 96.4% sensitivity and 82.1% specificity in distinguishing patients with pancreatic cancer from healthy subjects.ConclusionsThe authors observed associations between variations of patients' salivary microbiota with pancreatic cancer and chronic pancreatitis. This report also provides proof of salivary microbiota as an informative source for discovering non-invasive biomarkers of systemic diseases. | Publication | 21994333 (PMID) 10.1136/gutjnl-2011-300784 (DOI) |
Hu S, Vissink A, Arellano M, Roozendaal C, Zhou H, Kallenberg CG, Wong DT. Identification of autoantibody biomarkers for primary Sjögren's syndrome using protein microarrays. (2011) Proteomics 11: 1499-507 Show Abstract · Added October 15, 2011Sjögren’s syndrome (SS) is a chronic, progressive autoimmune disease primarily affecting women. Diagnosis of SS requires an invasive salivary gland tissue biopsy and a long delay from the start of the symptoms to final diagnosis has been frequently observed. In this study,we aim to identify salivary autoantibody biomarkers for primary SS (pSS) using a protein microarray approach. Immune-response protoarrays were used to profile saliva autoantibodies from patients with pSS (n = 514), patients with systemic lupus erythematosus(SLE, n = 513), and healthy control subjects (n = 513). We identified 24 potential autoantibody biomarkers that can discriminate patients with pSS from both patients with SLE and healthy individuals. Four saliva autoantibody biomarkers, anti-transglutaminase, anti-histone, anti-SSA, and anti-SSB, were further tested in independent pSS (n = 534), SLE (n = 534), and healthy control (n = 534) subjects and all were successfully validated with ELISA. This study has demonstrated the potential of a high-throughput protein microarray approach for the discovery of autoantibody biomarkers. The identified saliva autoantibody biomarkers may lead to a clinical tool for simple, noninvasive detection of pSS at low cost. | Publication | 21413148 (PMID) 10.1002/pmic.201000206 (DOI) |
Zhang L, Xiao H, Karlan S, Zhou H, Gross J, Elashoff D, Akin D, Yan X, Chia D, Karlan B, Wong DT. Discovery and preclinical validation of salivary transcriptomic and proteomic biomarkers for the non-invasive detection of breast cancer. (2010) PLoS One 5: e15573 Show Abstract · Added October 15, 2011A sensitive assay to identify biomarkers using non-invasively collected clinical specimens is ideal for breast cancer detection. While there are other studies showing disease biomarkers in saliva for breast cancer, our study tests the hypothesis that there are breast cancer discriminatory biomarkers in saliva using de novo discovery and validation approaches. This is the first study of this kind and no other study has engaged a de novo biomarker discovery approach in saliva for breast cancer detection. In this study, a case-control discovery and independent preclinical validations were conducted to evaluate the performance and translational utilities of salivary transcriptomic and proteomic biomarkers for breast cancer detection. | Publication | 21217834 (PMID) PMC3013113 (PMCID) 10.1371/journal.pone.0015573 (DOI) |
Hu S, Gao K, Pollard R, Arellano-Garcia M, Zhou H, Zhang L, Elashoff D, Kallenberg CG, Vissink A, Wong DT. Preclinical validation of salivary biomarkers for primary Sjögren's syndrome. (2010) Arthritis Care Res (Hoboken) 62: 1633-8 Show Abstract · Added October 15, 2011Sjögren’s syndrome (SS) is a systemic autoimmune disease with a variety of presenting symptoms that may delay its diagnosis. We previously discovered a number of candidate salivary biomarkers for primary SS using both mass spectrometry and expression microarray analysis. In the current study, we aimed to verify these candidate biomarkers in independent patient populations and to evaluate their predictive values for primary SS detection. | Publication | 20617533 (PMID) PMC2995446 (PMCID) 10.1002/acr.20289 (DOI) |
Palanisamy V, Sharma S, Deshpande A, Zhou H, Gimzewski J, Wong DT. Nanostructural and transcriptomic analyses of human saliva derived exosomes. (2010) PLoS One 5: e8577 Show Abstract · Added October 15, 2011Exosomes, derived from endocytic membrane vesicles are thought to participate in cell-cell communication and protein and RNA delivery. They are ubiquitous in most body fluids (breast milk, saliva, blood, urine, malignant ascites, amniotic, bronchoalveolar lavage, and synovial fluids). In particular, exosomes secreted in human saliva contain proteins and nucleic acids that could be exploited for diagnostic purposes. To investigate this potential use, we isolated exosomes from human saliva and characterized their structural and transcriptome contents. | Publication | 20052414 (PMID) PMC2797607 (PMCID) 10.1371/journal.pone.0008577 (DOI) |
Zhang L, Farrell JJ, Zhou H, Elashoff D, Akin D, Park NH, Chia D, Wong DT. Salivary transcriptomic biomarkers for detection of resectable pancreatic cancer. (2010) Gastroenterology 138: 949-57.e1-7 Show Abstract · Added October 15, 2011Lack of detection technology for early pancreatic cancer invariably leads to a typical clinical presentation of incurable disease at initial diagnosis. New strategies and biomarkers for early detection are sorely needed. In this study, we have conducted a prospective sample collection and retrospective blinded validation to evaluate the performance and translational utilities of salivary transcriptomic biomarkers for the noninvasive detection of resectable pancreatic cancer. | Publication | 19931263 (PMID) PMC2831159 (PMCID) 10.1053/j.gastro.2009.11.010 (DOI) |
Park NJ, Zhou H, Elashoff D, Henson BS, Kastratovic DA, Abemayor E, Wong DT. Salivary microRNA: discovery, characterization, and clinical utility for oral cancer detection. (2009) Clin Cancer Res 15: 5473-7 Show Abstract · Added November 19, 2011We have previously shown that a transcriptome is found in saliva and subpanels of these mRNAs can be used as oral cancer biomarkers. In this study, we measured the presence of microRNAs (miRNA) in saliva and determined their potential as an additional set of oral cancer biomarkers. | Publication | 19706812 (PMID) PMC2752355 (PMCID) 10.1158/1078-0432.CCR-09-0736 (DOI) |
Gao K, Zhou H, Zhang L, Lee JW, Zhou Q, Hu S, Wolinsky LE, Farrell J, Eibl G, Wong DT. Systemic disease-induced salivary biomarker profiles in mouse models of melanoma and non-small cell lung cancer. (2009) PLoS One 4: e5875 Show Abstract · Added October 15, 2011Saliva (oral fluids) is an emerging biofluid poised for detection of clinical diseases. Although the rationale for oral diseases applications (e.g. oral cancer) is intuitive, the rationale and relationship between systemic diseases and saliva biomarkers are unclear. | Publication | 19517020 (PMID) PMC2691577 (PMCID) 10.1371/journal.pone.0005875 (DOI) |
Wei F, Patel P, Liao W, Chaudhry K, Zhang L, Arellano-Garcia M, Hu S, Elashoff D, Zhou H, Shukla S, Shah F, Ho CM, Wong DT. Electrochemical sensor for multiplex biomarkers detection. (2009) Clin Cancer Res 15: 4446-52 Show Abstract · Added October 15, 2011Multiplexing assay of biomarkers at the point-of-care is an elusive goal for molecular diagnostics. | Publication | 19509137 (PMID) PMC2799532 (PMCID) 10.1158/1078-0432.CCR-09-0050 (DOI) |
Hu S, Zhou M, Jiang J, Wang J, Elashoff D, Gorr S, Michie SA, Spijkervet FK, Bootsma H, Kallenberg CG, Vissink A, Horvath S, Wong DT. Systems biology analysis of Sjögren's syndrome and mucosa-associated lymphoid tissue lymphoma in parotid glands. (2009) Arthritis Rheum 60: 81-92 Show Abstract · Added October 15, 2011To identify key target genes and activated signaling pathways associated with the pathogenesis of Sjögren's syndrome (SS) by conducting a systems analysis of parotid glands manifesting primary SS or primary SS/mucosa-associated lymphoid tissue (MALT) lymphoma phenotypes. | Publication | 19116902 (PMID) PMC2718690 (PMCID) 10.1002/art.24150 (DOI) |
Hu S, Arellano M, Boontheung P, Wang J, Zhou H, Jiang J, Elashoff D, Wei R, Loo JA, Wong DT. Salivary proteomics for oral cancer biomarker discovery. (2008) Clin Cancer Res 14: 6246-52 Show Abstract · Added October 15, 2011This study aims to explore the presence of informative protein biomarkers in the human saliva proteome and to evaluate their potential for detection of oral squamous cell carcinoma (OSCC). | Publication | 18829504 (PMID) PMC2877125 (PMCID) 10.1158/1078-0432.CCR-07-5037 (DOI) |
Hu Z, Zimmermann BG, Zhou H, Wang J, Henson BS, Yu W, Elashoff D, Krupp G, Wong DT. Exon-level expression profiling: a comprehensive transcriptome analysis of oral fluids. (2008) Clin Chem 54: 824-32 Show Abstract · Added October 15, 2011The application of global gene expression profiling to saliva samples is hampered by the presence of partially fragmented and degraded RNAs that are difficult to amplify and detect with the prevailing technologies. Moreover, the often limited volume of saliva samples is a challenge to quantitative PCR (qPCR) validation of multiple candidates. The aim of this study was to provide proof-of-concept data on the combination of a universal mRNA-amplification method with exon arrays for candidate selection and a multiplex preamplification method for easy validation. | Publication | 18356245 (PMID) PMC2799536 (PMCID) 10.1373/clinchem.2007.096164 (DOI) |