Faina Linkov

Early Detection of Head and Neck Cancer: Development of a Novel Screening Tool Using Multiplexed Immunobead-Based Biomarker Profiling

Squamous cell carcinoma of the head and neck (SCCHN) is an aggressive disease that has been linked to altered immune, inflammatory, and angiogenesis responses. A better understanding of these aberrant responses might improve early detection and prognosis of SCCHN and provide novel therapeutic targets. Previous studies examined the role of multiplexed serum biomarkers in small cohorts or SCCHN sera. We hypothesized that an expanded panel comprised of multiple cytokines, chemokines, growth factors, and other tumor markers, which individually may show some promising correlation with disease status, might provide higher diagnostic power if used in combination. Thus, we evaluated a novel multianalyte LabMAP profiling technology that allows simultaneous measurement of multiple serum biomarkers. Concentrations of 60 cytokines, growth factors, and tumor antigens were measured in the sera of 116 SCCHN patients before treatment (active disease group), 103 patients who were successfully treated (no evidence of disease group), and 117 smoker controls without evidence of cancer. The multimarker panel offering the highest diagnostic power was comprised of 25 biomarkers, including epidermal growth factor, epidermal growth factor receptor, interleukin (IL)-8, tissue plasminogen activator inhibitor-1, α-fetoprotein, matrix metalloproteinase-2, matrix metalloproteinase-3, IFN-α, IFN-γ, IFN-inducible protein-10, regulated on activation, normal T-cell expressed and secreted (RANTES), macrophage inflammatory protein-1α, IL-7, IL-17, IL-1 receptor-α, IL-2 receptor, granulocyte colony-stimulating factor, mesothelin, insulin-like growth factor binding protein 1, E-selectin, cytokeratin-19, vascular cell adhesion molecule, and cancer antigen-125. Statistical analysis using an ADE algorithm resulted in a sensitivity of 84.5%, specificity of 98%, and 92% of patients in the active disease group correctly classified from a cross-validation serum set. The data presented show that simultaneous testing using a multiplexed panel of serum biomarkers may present a promising new approach for the early detection of head and neck cancer. (Cancer Epidemiol Biomarkers Prev 2007;16(1):102–7)

Reproducibility of serum cytokines and growth factors

BACKGROUND In most studies, circulating biomarkers are usually assessed from a single sample, assuming that this single measurement represents the long-term biomarker status of the individual. Such an assumption is rarely tested although it may not be valid for all biomarkers. The objective of this study was to investigate the temporal reproducibility of a panel of cytokines and growth factors. METHODS Thirty-five postmenopausal women with two annual visits and 30 premenopausal women with three annual visits were randomly selected from the participants in an existing prospective cohort. A total of 23 serum cytokines, nine growth factors and C-reactive protein (CRP) were measured using the Luminex xMap technology. In addition, for eight biomarkers, regular and high sensitivity (hs) assays were compared. RESULTS The biomarkers with adequate (>60%) detection rates and acceptable (> or =0.55) intra-class correlation coefficients (ICCs) were: hsIL-1beta, IL-1RA, hsIL-2, hsIL-4, hsIL-5, hsIL-6, hsIL-10, IL-12p40, hsIL-12p70, hsTNF-alpha, TNF-R1, TNF-R2, CRP, HGF, NGF, and EGFR. The remaining biomarkers either had low temporal reproducibility or were undetectable in more than 40% of samples. CONCLUSIONS The results suggest that 16 of the 41 biomarkers measured with Luminex technology showed sufficient sensitivity and temporal reproducibility in sera.

USA Endometrial Cancer Projections to 2030: should we be concerned?

AIM As the incidence of endometrial cancer (EC) increased considerably since 2007, this study aimed to project the burden of EC to the year 2030. METHODS Multivariate linear regression was used to project EC incidence by modeling trends in EC incidence from 1990 to 2013, while accounting for temporal changes in obesity, hysterectomy and smoking. RESULTS The best-fitting model predicting EC rates included a time effect plus effects for hysterectomy (12-year lag), severe obesity (3-year lag) and smoking (9-year lag). The best-fitting model projected an increase to 42.13 EC cases per 100,000 by the year 2030, a 55% increase over 2010 EC rates. CONCLUSION The projected increase of EC over next 16 years indicates the need for close monitoring of EC trends.

Endometrial hyperplasia, endometrial cancer and prevention: Gaps in existing research of modifiable risk factors

PURPOSE Endometrial carcinoma is the most common cancer of the lower female genital tract in Europe and the United States. Faced with the growing incidence of endometrial cancer in Europe and around the world, scientists, doctors and public health professionals are becoming more concerned with identifying effective preventive measures for this condition. This review paper presents the existing knowledge about modifiable risk factors leading to endometrial hyperplasia and endometrial cancer and highlights the need for more studies in this area. DESIGN/METHODS Extensive literature review of modifiable risk factors for endometrial cancer and endometrial hyperplasia has been performed. Additionally, biomarker approaches to cancer monitoring, existing therapies for endometrial hyperplasia and factors affecting patient survival are reviewed. RESULTS Obesity and inactivity are two of the major risk factors associated with the development of endometrial cancer and endometrial hyperplasia. Other modifiable risk factors include dietary habits, exercise and the use of hormonal therapy. Similar factors, along with cancer biomarkers, may play an important role in the early detection of endometrial cancer and survival after the diagnosis. The majority of these factors fit well with the unopposed oestrogen theory. Diet and exercise programmes are currently not integrated into a standard treatment programmes for patients with endometrial hyperplasia or endometrial cancer. CONCLUSIONS More studies are needed to investigate modifiable risk factors for endometrial cancer and endometrial hyperplasia. Existing therapies for endometrial hyperplasia target hormone imbalance, which is just one aspect of endometrial cancer development. Next generation therapies for endometrial cancer and endometrial hyperplasia patients should include diet, exercise and weight loss plans, which would target other modifiable aspects of endometrial cancer risk.

Papyrus to PowerPoint (P 2 P): metamorphosis of scientific communication

Scientific communication is in the process of metamorphosis. Will it change into a dung beetle or into a beautiful butterfly? Here is one possibility that some might argue is as frightening as Kafkas story > “As Gregor Samsa awoke from unsettling dreams one morning, he found himself transformed in his bed into a monstrous bug.” > > Kafka, Metamorphosis In 1995 we questioned the hallowed tenets of paper journals. We wrote a series of articles, beginning with “The death of biomedical journals,” suggesting the death knell for paper journals.1–3 Delamothe echoed our conclusions that “The burgeoning world wide web … makes it inevitable that new systems of disseminating research will replace or at least supplement journals.”4 ![][1] #### Summary points Traditional peer reviewed journals are becoming obsolete We are experiencing a dramatic metamorphosis of the tools of scientific communication The prima lingua of scientific communication is PowerPoint Our search for the optimal information exchange method in science leads to P2P The response was Kafkaesque, reminding us of the quote from Penal Colony “It is an exceptional apparatus” so do not question it. The “journal” apparatus shows that little of the fibre of journals has been scientifically evaluated. Are journals an efficient, scientific, “just in time” process? It is impossible to answer. For 300 years there has been no evidence based evaluation of the journal process. For example, there is virtually no research on the quality of learning from journals, whether IMRD (introduction, methods, results, discussion) optimises learning, or if traditional peer review is the best system. To quote Goldbeck-Wood, “But if peer review is so central to the process by which scientific knowledge become canonised, it is ironic that science has little to say as to whether it works.”5 This applies to all phases of the journal process. Is a metamorphosis in … [1]: /embed/graphic-1.gif

Multiple biomarker panels for early detection of ovarian cancer

Ovarian cancer is the eighth most common cause of cancer mortality in women. It is diagnosed in more than 20,000 women in the USA each year and approximately 15,000 women die of the disease annually. The majority of patients are diagnosed with advanced-stage ovarian cancer, as this deadly disease causes minimal and nonspecific symptoms until late in the course of the disease. No standardized screening test exists to reliably detect ovarian cancer. Cancer antigen (CA)-125 is a protein antigen found at abnormally high levels in the blood of many women with ovarian cancer. Most healthy women have CA-125 levels of below 35 units/microl of blood serum. However, a number of noncancerous conditions can cause elevated CA 125 levels, and many women with early-stage ovarian cancer have normal CA-125 levels. Owing to these limitations, this test is not recommended for routine screening in women who are not at high risk or who do not have specific symptoms of the disease. Currently, many researchers are focusing on simultaneous examination of multiple markers to increase sensitivity of the screening test for early detection of ovarian cancer. Analysis of the current literature shows that combining several biomarkers dramatically improves sensitivity of CA-125 in ovarian cancer patients. This article provides a comprehensive overview of existing studies in the area of multimarker panel development for the early detection and monitoring of ovarian cancer. Our literature review demonstrates that a multimarker approach for the generation of a prototype assay for early detection of ovarian cancer has a great potential to lead to the development of a screening test for this disease.

Reliability of tumor markers, chemokines, and metastasis-related molecules in serum

There is a growing interest in the role that cancer biomarkers, metastasis-related molecules, and chemokines may play in the development and progression of various cancers. However, few studies have addressed the reliability of such biomarkers in healthy individuals over time. The objective of this study was to investigate the temporal reliability of multiple proteins in serum samples from healthy women who donated blood over successive years. Thirty five, postmenopausal women with two, repeated annual visits, and thirty, premenopausal women with three, repeated annual visits were randomly selected among eligible subjects from an existing, prospective cohort. Multiplexing Luminex xMAPTM technology was used to measure the levels of 55 serum proteins representing cancer antigens, chemokines, angiogenic and anti-angiogenic factors, proteases, adipokines, apoptotic molecules, and other markers in these women. The biomarkers with high detection rates (> 60%) and acceptable reliability (intraclass correlation coefficient, ICCs > or = 0.55) using xMAPTM method were: cancer antigens: AFP, CA 15-3, CEA, CA-125, SCC, SAA; growth factors/related molecules: ErbB2, IGFBP-1; proteases and adhesion molecules: MMP-1, 8, 9, sE-selectin, human kallikreins (KLK) 8,10, ICAM-1, VCAM-1, chemokines: fractalkine, MCP-1,2, RANTES, MIP-1alpha, MIP-1beta, Eotaxin, GRO-alpha, IP-10; inhibitors of angiogenesis: angiostatin and endostatin; adipokines leptin and resistin; apoptotic factor: Fas, and other proteins mesothelin, myeloperoxidase (MPO), and PAI-1. The rest of the biomarkers under investigation either had ICCs less than 0.55 or had low levels of detection (< 60%). These included cancer antigens: CA 19-9, CA 72-4, MICA, S100, TTR, ULBP1, ULBP2, ULBP3; proteases: MMP 2, 3, 7, 12, 13; chemokines: MCP-3, MIF, MIG; adipokines: leptin and resistin; apoptotic factors: FasL, DR5, Cyfra 21-1; and inhibitors of angiogenesis and other markers: thrombospondin and heat shock protein (HSP) 27. In conclusion, 34 out of the 55 biomarkers investigated were present in detectable levels in > 60% of the samples, and with an ICC > or = 0.55, indicating that a single serum measurement can be used in prospective epidemiological studies using the xMAPTM method.

Comparison of survival outcomes between patients with malignant mixed mullerian tumors and high-grade endometrioid, clear cell, and papillary serous endometrial cancers.

Introduction: Malignant mixed mullerian tumors (MMMTs) are an aggressive subtype of endometrial cancer (EC). Previous studies compare survival between high-grade endometrioid (EM), clear cell (CC), and papillary serous (PS) ECs; yet few studies compare MMMTs to these aggressive subtypes. The goal of this study was to compare recurrence-free survival (RFS), disease-specific survival (DSS), and overall survival (OS) among EC subtypes. Methods: We conducted a retrospective cohort study of EC cases treated at Magee-Womens Hospital between 1996 and 2008. Kaplan-Meier estimates of RFS, DSS, and OS as well as and log-rank tests were used to compare survival distributions between histologic subtypes. Cox regression was used to estimate hazard ratios for histologic subtypes, adjusted for other significant prognostic factors. Interactions between histologic subtype and prognostic factors were examined to assess effect modification. Results: This cohort included 81 MMMT (15%), 254 high-grade EM (46%), 73 CC (13%), and 147 PS (26%) cases. Compared to high-grade EM (6%) and CC (7%) cases, relatively more MMMT (12%) and PS (12%) cases were nonwhite. Stage differed significantly among the subtypes, with 36%, 34%, 37%, and 51% of MMMT, high-grade EM, CC, and PS cases, respectively, diagnosed at advanced late stage (P < 0.001). Kaplan-Meier curves and log-rank tests showed similar RFS, DSS, and OS between MMMT, high-grade EM, CC, and PS cases stratified by stage. In adjusted Cox regression models, RFS and DSS were not significantly different between MMMT and other subtypes. High-grade EM cases had a significantly better OS compared to MMMT cases (HR, 0.63; 95% confidence interval [CI], 0.41-0.98). Conclusions: This is the first retrospective study to suggest that certain survival outcomes are similar among MMMT, high-grade EM, CC, and PS subtypes. Other large-scale studies are needed to confirm these findings.

Hormone receptor expression patterns in the endometrium of asymptomatic morbidly obese women before and after bariatric surgery

OBJECTIVE Obesity increases risk for endometrial neoplasia, but neither the pathophysiology nor the effects of weight loss on the risk are well established. We attempted to characterize the molecular profile of the endometrium of asymptomatic women with morbid obesity before and following bariatric surgery-induced weight loss. METHODS 59 asymptomatic, morbidly obese women underwent endometrial sampling before bariatric surgery; 46 (78%) of these returned one year later for re-biopsy (median weight loss of 41kg). Duplicate samples from these specimens were scored for expression of estrogen receptor (ER), progesterone receptor (PR), androgen receptor (AR), and Ki-67 by two independent, blinded pathologists using an H-score [staining intensity (0-3)×(percent of tissue involved)]. RESULTS The prevalence of hyperplasia pre-operatively was 7% overall and 10% among patients not on an anti-estrogen. ER H-scores were similar before and after surgery overall (median 190 and 196 respectively, p=0.82), but patients with hyperplasia had higher pre-operative H-scores (median 256, p<0.001) and experienced greater H-score drops, than those without hyperplasia (-112 vs +50, p=0.028). In two patients with persistent hyperplasia at one year, ER H-scores fell to levels that were similar to those without pathology. One patient who developed hyperplasia during the study period had a rising ER H-score. Patients with hyperplasia had higher median PR H-scores pre-operatively (284 vs 188, p=0.01), which normalized through greater drops (75 vs 0, p=0.053). AR H-scores dropped significantly after surgery (13 vs 2, p=0.015), but were similar between patients with and without hyperplasia (p=0.33). Weight loss did not affect Ki-67 proliferation index. CONCLUSION Asymptomatic morbidly obese patients have a high prevalence of occult hyperplasia, characterized by relatively high hormone receptor expression. These profiles appear to normalize with weight loss and in advance of pathologically identifiable changes. These data suggest a potential role for screening this population as well as the possibility that weight loss may be a valid treatment strategy for risk reduction.

Reproducibility of Serum Pituitary Hormones in Women

Endogenous pituitary hormones are commonly used in clinical and epidemiologic studies and some of them are thought to influence the risk of several diseases in women. In most studies, endogenous levels of pituitary hormones are usually assessed at a single point in time, assuming that this single measurement represents the long-term biomarker status of the individual. Such an assumption is rarely tested and may not always be valid. This study examined the reproducibility of the following pituitary hormones: adrenocorticotropic hormone (ACTH), growth hormone, follicle-stimulating hormone (FSH), luteinizing hormone (LH), thyroid-stimulating hormone (TSH), and prolactin, measured using the Luminex xMap method in sera of healthy premenopausal and postmenopausal women. The study included 30 premenopausal women with three yearly samples and 35 postmenopausal women with two repeated yearly samples randomly selected from an existing prospective cohort. Analysis of intraclass correlation coefficients suggested higher reproducibility in postmenopausal women compared with premenopausal women for the following hormones: FSH (0.72 and 0.37, respectively), LH (0.83 and 0.44, respectively), and growth hormone (0.60 and 0.35, respectively). The intraclass correlation coefficients were relatively high and similar between postmenopausal and premenopausal women for ACTH (0.95 and 0.94, respectively), TSH (0.85 and 0.85, respectively), and prolactin (0.72 and 0.69, respectively). This study found that serum concentrations of FSH, LH, and growth hormone are stable in postmenopausal women and that ACTH, TSH, and prolactin are stable in both premenopausal and postmenopausal women, suggesting that a single measurement may reliably categorize average levels over at least a 2-year period. (Cancer Epidemiol Biomarkers Prev 2008;17(8):1880–3)