Seraina Schmid

Hypermethylation of Tumor Suppressor Genes Involved in Critical Regulatory Pathways for Developing a Blood-Based Test in Breast Cancer

Background Aberrant DNA methylation patterns might be used as a biomarker for diagnosis and management of cancer patients. Methods and Findings To achieve a gene panel for developing a breast cancer blood-based test we quantitatively assessed the DNA methylation proportion of 248 CpG sites per sample (total of 31,248 sites in all analyzed samples) on 10 candidate genes (APC, BIN1, BMP6, BRCA1, CST6, ESR-b, GSTP1, P16, P21 and TIMP3). The number of 126 samples consisting of two different cohorts was used (first cohort: plasma samples from breast cancer patients and normal controls; second cohort: triple matched samples including cancerous tissue, matched normal tissue and serum samples). In the first cohort, circulating cell free methylated DNA of the 8 tumor suppressor genes (TSGs) was significantly higher in patients with breast cancer compared to normal controls (P<0.01). In the second cohort containing triple matched samples, seven genes showed concordant hypermethylated profile in tumor tissue and serum samples compared to normal tissue (P<0.05). Using eight genes as a panel to develop a blood-based test for breast cancer, a sensitivity and specificity of more than 90% could be achieved in distinguishing between tumor and normal samples. Conclusions Our study suggests that the selected TSG panel combined with the high-throughput technology might be a useful tool to develop epigenetic based predictive and prognostic biomarker for breast cancer relying on pathologic methylation changes in tumor tissue, as well as in circulation.

Levels of circulating cell-free nuclear and mitochondrial DNA in benign and malignant ovarian tumors

OBJECTIVE: To analyze the levels of circulating cell-free nuclear DNA and circulating cell-free mitochondrial DNA in patients with benign and malignant ovarian tumors using a gold-standard assay and to investigate whether quantitative alterations of the circulating cell-free species have values in the management of the patients. METHODS: One hundred four patients were recruited for this study. We developed a quantitative, multiplex polymerase chain reaction to measure the levels of circulating cell-free nuclear DNA and circulating cell-free mitochondrial DNA in serum and plasma of patients with epithelial ovarian cancer, benign epithelial ovarian tumors, or endometriosis. The levels of the circulating cell-free DNA were compared with those of a healthy, age-matched control group. RESULTS: The patients with epithelial ovarian cancer had significantly higher amounts of circulating cell-free nuclear DNA and circulating cell-free mitochondrial DNA in plasma compared with the healthy control group (mean of nuclear DNA 10,723/2,591 and mean of mitochondrial DNA 4,918,978/2,294,264, P=.009 and 0.022, respectively) and with the other group with benign ovarian diseases (mean of nuclear DNA 10,723/2,965 and mean of mitochondrial DNA 4,918,978/1,597,551, P=.027 and 0.002, respectively). However, no relationship between levels of the circulating cell-free DNA and the pathological parameters as well as CA 125 measurement in patients with epithelial ovarian cancer was found. A significant difference between the epithelial ovarian cancer and endometriosis group was found in circulating cell-free mitochondrial DNA but not in circulating cell-free nuclear DNA (mean of mitochondrial DNA 4,918,978/2,273,988 and mean of nuclear DNA 10,723/3,291, P=.013 and 0.105, respectively). CONCLUSION: Elevated levels of circulating cell-free nuclear DNA and circulating cell-free mitochondrial DNA in epithelial ovarian cancer may have diagnostic value. Our finding suggests that the circulating molecules might be potential biomarkers in the disease. LEVEL OF EVIDENCE: III

Circulating cell-free DNA as a potential biomarker for minimal and mild endometriosis

It has recently been reported that high concentrations of circulating cell-free (ccf) nucleic acids in plasma and serum could be used as biomarkers for non-invasive monitoring a wide variety of malignant and benign proliferations and inflammatory conditions. Endometriosis is one of the most common benign gynaecological proliferations with inflammatory activation in premenopausal women. Real-time multiplex polymerase chain reaction was used for synchronized quantification of the glyceraldehyde-3-phosphate dehydrogenase gene sequence in nuclear DNA (nDNA) and the ATP synthase-8 gene sequence in mitochondrial DNA (mtDNA). DNA was extracted from 500 microl serum and plasma of 19 cases with endometriosis to measure the total amount of ccf nDNA and ccf mtDNA. The concentration of ccf nDNA in plasma was significantly higher in the endometriosis group than in the control group (P = 0.046). The cut-off value selected by a receiver operating characteristic curve could provide a sensitivity of 70% and a specificity of 87% to discriminate between the minimal or mild cases and normal controls. The finding of significantly increased concentrations of ccf nDNA in plasma of patients with endometriosis suggests that ccf nDNA might be a potential biomarker for developing non-invasive diagnostic test in endometriosis.

Levels of circulating cell-free serum DNA in benign and malignant breast lesions.

Purposes of the study: We analyzed circulating cell-free DNA in the serum of patients with benign and malignant breast disease and in healthy individuals to determine its diagnostic value. BASIC PROCEDURES Serum samples were obtained from 50 healthy individuals, 33 patients with malignant breast disease and 32 patients with benign breast disease. Circulatory DNA was extracted from serum samples. Cell-free DNA was quantified by real-time quantitative PCR for the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene. Tissue samples from patients with malignant and benign breast lesions were histopathologically examined. MAIN FINDINGS The mean levels of circulating cell-free DNA in serum samples were 41,149 genome equivalents (GE)/mL in patients with malignant disease, 30,826 GE/mL in patients with benign disease, and 13,267 GE/mL in healthy individuals. Healthy individuals had significantly lower levels of cell-free DNA than patients with malignant or benign breast disease (p=0.001, p=0.031). No significant difference was observed between malignant and benign disease. There was a correlation between cell-free DNA levels and tumor size but not with other tumor characteristics. PRINCIPAL CONCLUSION Our results suggest that levels of circulating cell-free DNA in serum could have diagnostic value to discriminate between healthy individuals and patients with breast lesions but not between patients with malignant and benign breast lesions.PURPOSES OF THE STUDY We analyzed circulating cell-free DNA in the serum of patients with benign and malignant breast disease and in healthy individuals to determine its diagnostic value. BASIC PROCEDURES Serum samples were obtained from 50 healthy individuals, 33 patients with malignant breast disease and 32 patients with benign breast disease. Circulatory DNA was extracted from serum samples. Cell-free DNA was quantified by real-time quantitative PCR for the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene. Tissue samples from patients with malignant and benign breast lesions were histopathologically examined. MAIN FINDINGS The mean levels of circulating cell-free DNA in serum samples were 41,149 genome equivalents (GE)/mL in patients with malignant disease, 30,826 GE/mL in patients with benign disease, and 13,267 GE/mL in healthy individuals. Healthy individuals had significantly lower levels of cell-free DNA than patients with malignant or benign breast disease (p=0.001, p=0.031). No significant difference was observed between malignant and benign disease. There was a correlation between cell-free DNA levels and tumor size but not with other tumor characteristics. PRINCIPAL CONCLUSION Our results suggest that levels of circulating cell-free DNA in serum could have diagnostic value to discriminate between healthy individuals and patients with breast lesions but not between patients with malignant and benign breast lesions.

Impact of body mass index on compliance and persistence to adjuvant breast cancer therapy

Several authors found that the prognosis of overweight and obese breast cancer (BC) patients was lower than that of normal weight patients. We present the first study which evaluates the impact of body mass index (BMI) on compliance (i.e. to start a recommended therapy) and persistence to adjuvant BC therapy. An unselected cohort of 766 patients (≤75 years) diagnosed from 1997 to 2009 was analyzed in relevance to the four adjuvant therapy modalities: (A) radiation, (B) chemotherapy, (C) therapy with trastuzumab, and (D) endocrine therapy. With respect to compliance, multivariate analyses calculated Odds ratios (ORs) >1 for increased BMI in all four therapy modalities, i.e. increased BMI had a positive influence on compliance. The results were significant for radiotherapy (OR,2.37;95%CI,1.45-3.88;p < 0.001) and endocrine therapy (OR,1.92;95%CI,1.21-3.04;p = 0.002) and showed a trend in chemotherapy (OR,1.42;95%CI,0.97-2.08;p = 0.063). Analyzing persistence, increasing BMI had ORs <1 for chemotherapy and therapy with trastuzumab, both not reaching statistical significance. For endocrine therapy, increasing BMI was a significant predictor for persistence (OR,1.35;95%CI,1.08-1.80;p = 0.042). Failure of compliance and persistence to adjuvant therapy does not pose a contributing factor for the observed unfavorable prognosis in overweight/obese BC patients. In most therapy modes, patients with increasing BMI demonstrated a higher motivation and perseverance to the recommended treatment.

Eligibility, compliance and persistence of extended adjuvant endocrine therapy for breast cancer

Abstract Background. Extended therapy (ET) beyond the standard five years of tamoxifen-containing treatment is a widely discussed therapy option in adjuvant endocrine breast cancer (BC) therapy which might offer an opportunity for further protection against late relapses. In this study we evaluated eligibility, compliance and persistence of extended adjuvant endocrine BC therapy. Patients and methods. Data concerning all BC patients (≤75 years) who initiated endocrine adjuvant therapy between 1999 and 2005 (n = 286) was analyzed. Results. One hundred and thirty-eight patients were valid candidates for an ET according current guidelines; this represents 48.3% of the individuals who started endocrine therapy five years ago. Of these, 89 (64.5%) received a corresponding offer/recommendation by their treating physicians. Advanced age (p = 0.002), favorable disease stage (p = 0.011), and follow-up at a general practitioner (p < 0.001) were significant factors where a recommendation for an ET was not made. Of the 89 patients who were offered an ET, 64 followed this proposal (compliance: 84.7%). Eighteen patients (28.1%) were non-persistent to the ET; therapy-related adverse effects were the main reason for discontinuation. Sixteen patients received an ET beyond current guidelines (tamoxifen or an aromatase inhibitor alone was given longer than five years); this represents 11.0% of all patients who completed five years of endocrine therapy. Conclusions. Only a minority of the patients who started an endocrine therapy were actually eligible for an ET. Patients who were offered/recommended an ET had a high rate of compliance and persistence. Efforts should be made to make sure that all physicians, above all general practitioners, who are involved in the treatment of BC patients, are provided with current therapy guidelines as to guarantee an optimal patient management.

Cannabinoid hyperemesis syndrome: an underreported entity causing nausea and vomiting of pregnancy

IntroductionIn the western world, cannabis is the most widely used drug of abuse. Cannabinoid hyperemesis syndrome, which seems to be a rare paradoxical reaction in individuals with a particular predisposition, is characterized by cyclic severe nausea and vomiting in long-term cannabis users. While the symptoms are unresponsive to antiemetic drugs, compulsive hot baths result in a considerable symptom relief.MethodsWe report the first case of cannabinoid hyperemesis syndrome in pregnancy. A 26-year-old patient was admitted to our clinic in the 10th week of gestation.ConclusionBefore undertaking time-consuming and expensive medical examinations to rule out other medical reasons for therapy-resistant hyperemesis in pregnancy, obstetricians should determine whether compulsive bathing or showering provides symptomatic relief and ask specific questions regarding possible/suspected cannabis consumption.

Suicide in breast cancer patients: An individual-centered approach provides insight beyond epidemiology

Abstract Background. Epidemiologic studies have identified increased suicide rates among breast cancer (BC) patients. The population-based approach, however, has considerable methodic shortcomings. None of the studies have been carried out in a prospective manner and none reported suicide rates from a country in which physician-assisted suicide (PAS) is legal. Patients and methods. All cases recorded by a prospective Swiss BC database during a 17-year period (1990–2006; n = 1165) were analyzed. Using an individual-centered approach, the cases of women who committed suicide are reported according to the psychological autopsy method. Results. In six patients (0.5%; 5.1/1.000 patients), suicidal death was identified. In four patients, suicide was committed during late stages of metastatic BC. In two cases, comorbid conditions were associated with suicide. Three women chose PAS. Conclusion. The individual-centered approach is a well-suited innovative concept to increase the knowledge regarding the relationship between cancer and suicide. We found a two to seven times higher suicide rate than those reported in epidemiologic studies. The population-based approach can barely elucidate the immense variety of one of the most personal decisions: the act of intentionally ending ones own life. These studies suffer from systematic failure of analysis since they did not a) consider the potential confounding role of comorbid medical and/or psychiatric conditions, and b) report in which disease stage suicide was committed, since the decisive disease-related event whether and when metastatic disease occurred was not recorded. Furthermore, epidemiologic data stems from countries in which PAS is prohibited and therefore not included in official statistics. This grey area of medicine accounts for a greater scope of underreporting than had previously been assumed.

Overweight, obesity, and breast cancer screening: results from the 2012 Swiss Health Survey.

Obesity is associated with poor breast cancer (BC) prognosis. Larger tumor size and more advanced disease stage at diagnosis could partly explain this outcome and nonadherence of obese women to BC screening may play a role. We examined the relationship between BMI (kg/m2) and the use of mammography in Switzerland as well as separately in the German-speaking part with mainly opportunistic screening and in the French-speaking part with organized programs. We analyzed the data of 50–69-year-old women (n=3121) of the Swiss Health Survey 2012. Study participants were classified as underweight (BMI<18.5), normal weight (BMI 18.5–24.9), overweight (BMI 25–29), or obese (BMI≥30). Outcome measures were dichotomized into 2 years or less since most recent mammography versus more than 2 years or never. We carried out multivariable logistic regression analyses, adjusting for sociodemographics, lifestyle, and self-perceived health. In Switzerland, 47% of women aged 50–69 years had had BC screening within the last 2 years, 35% of women in the German-speaking and 78% of women in the French-speaking part. In the total group, mammography use was higher in overweight than in normal-weight women (adjusted odds ratio 1.21, 95% confidence interval 0.98–1.49). Stratified by region, this effect was only maintained in women of the German-speaking part (adjusted odds ratio 1.41, 95% confidence interval 1.08–1.85; P-interaction<0.001). There were no differences in mammography attendance between normal-weight and obese and underweight women. In the 2012 Swiss Health Survey, overweight, obesity, and underweight were no barriers to mammography. In the German-speaking part, overweight women even attended mammography screening more often than normal-weight women.

Use of Palliative Endocrine Therapy in Patients with Hormone Receptor-Positive Distant Metastatic Breast Cancer: How Often, How Effective, How Long?

Background: This study provides real-world clinical evidence regarding palliative endocrine therapy (ET) in breast cancer (BC). The main questions to be answered were: how often and how long did patients receive ET? A particular aspect was the analysis of compliance and persistence with ET. Methods: An analysis of a nonselected/consecutive cohort of women with distant metastatic hormone receptor-positive BC (n = 205) was conducted. Results: In all, 165 patients (80.5%) received ET during the palliative disease course. The noncompliance rate was 1.5%. Sixty-seven patients (40.6%) had ET as the only antineoplastic therapy. The median number of therapy lines was 2, and the median duration was 18 months. The median metastatic disease survival (MDS) was 34 months. In patients who had an MDS of ≥9 months (n = 145; 87.9%), during 70.6% of the MDS time only ET had been administered. Patients who were naïve to ET more often had a good response to and a longer duration of palliative ET than those who were not. The nonpersistence rate was 4.3%. Conclusions: Excluding the few patients who had a rapidly progressive course, the disease was controlled for about 70% of the entire palliative disease course with ET alone. Only very few patients were nonpersistent with ET and consciously stopped a still effective, ongoing ET.