Markéta Protivánková

HOTAIR long non-coding RNA is a negative prognostic factor not only in primary tumors, but also in the blood of colorectal cancer patients

Colorectal cancer (CRC) is one of the main causes of death of neoplasia. Demand for predictive and prognostic markers to reverse this trend is increasing. Long non-coding RNA HOTAIR (Homeobox Transcript Antisense Intergenic RNA) overexpression in tumors was previously associated with poor prognosis and higher mortality in different carcinomas. We analyzed HOTAIR expression levels in tumor and blood of incident sporadic CRC patients in relation to their overall survival with the aim to evaluate surrogate prognostic marker for CRC. Tissue donor group consisted of 73 CRC patients sampled for tumor and normal tissue. Blood donor group was represented by 84 CRC patients compared with 40 healthy controls. Patients were characterized for tumor-node-metastasis stage, tumor grade, microsatellite instability and tumor penetration by stromal cells. HOTAIR levels were assessed by real-time quantitative PCR. CRC patients had higher HOTAIR expression in blood than healthy controls (P = 0.0001), whereas there was no difference in HOTAIR levels between tumor and adjacent mucosa of CRC patients. HOTAIR levels positively correlated between blood and tumor (R = 0.43, P = 0.03). High HOTAIR levels in tumors were associated with higher mortality of patients [Coxs proportional hazard, hazard ratio = 4.4, 95% confidence interval: 1.0-19.2, P = 0.046]. The hazard ratio was even higher when blood HOTAIR levels were taken into account (hazard ratio = 5.9, 95% confidence interval: 1.3-26.1, P = 0.019). Upregulated HOTAIR relative expression in primary tumors and in blood of CRC patients is associated with unfavorable prognosis. Our data suggest that HOTAIR blood levels may serve as potential surrogate prognostic marker in sporadic CRC.

Monitoring of minimal residual disease in acute myeloid leukemia with frequent and rare patient‐specific NPM1 mutations

Nucleophosmin (NPM1) mutations in exon 12 are the most common genetic alternation in cytogenetically normal AML (CN‐AML). Although mutation types A, B, and D represent the majority of cases, rare mutation variants of the NPM1 gene in individual patients do occur. In this study, we have evaluated a novel, DNA‐based real‐time quantitative polymerase chain reaction (RQ‐PCR) for the detection of three of the most commonly occurring mutations and for six rare patient‐specific mutation types, which represent 28% of all of the NPM1 mutations in our group of 25 CN‐AML patients. Furthermore, the prognostic relevance of NPM1‐based monitoring of minimal residual disease (MRD) in peripheral blood (PB), bone marrow (BM), and in specific cell subsets (CD34+, CD34−, CD34dim) of BM were evaluated. In 80% of the evaluable patients, a molecular relapse preceded a hematological relapse. Moreover, in this subset of patients, the molecular relapse occurred at a median of 97 days before the hematological relapse. Our compartment analysis showed a strong correlation between BM and PB (r = 0.907, P < 0.001) as well as a high copy number of mutated NPM1 in CD34+ BM cells. In conclusion, we have demonstrated applicability of our presented RQ‐PCR method for a large percentage of mutated NPM1 patients with CN‐AML as well as the usefulness for long‐term follow‐up monitoring of MRD and the prediction of hematological relapse. Am. J. Hematol., 2010.

Bone Markers in the Treatment of Cancer Related Bone Disease in Patients with Metastatic Breast Cancer

Bone metastases and treatment-induced osteoporosis are frequently the maincauses of morbidity in patients with malignancies. Monitoring the level of bone markers (markers of bone metabolism) has been fairly well mapped in osteoporosis, where medical procedures can be modified according to the kinetics of marker levels before an answer can be evaluated by densitometry and before the onset of fractures. In bone metastases the role of these levels is not clear. The metabolic markers of bone resorption under review in this set were s-Cross Laps (CTX) - a peptide that is a part of C - telopeptide, and N-terminal propeptide of collagen type 1 (P1NP). Material and methods: We monitored a group of 52 female patients with metastatic breastcancer. The patients received appropriate systemic treatment based on the immunohistochemistry of the tumor; the treatment consisted of hormone therapy or chemotherapy, and included parenteral bisphosphonates (ibandronate and zoledronic acid alternately). Routine biochemical tests and blood count done prior to the initiation of therapy also included taking laboratory markers of bone metabolism CTX and P1NP and measuring bone mineral density (according to T-score). These bone markers were then checked in three, six, nine and twelve months, and matched with the progress of the disease. Total monitoring time was fifteen months. Results: The patients in this set whose CTX value in the first sampling was less than 0.425 ran 8.5 times higher risk of death; the patients whose P1NP value reached more than 74 in the first collection ran 8.7 times higher risk of death. According to Cox proportional-hazards regression analysis for CTX, the significance level of p-value was 0.0452 and HR was 8.516 (95% CI 1.047 to 69.262), a difference which is not statistically significant. Regarding P1NP in Cox regression analysis, the significance level of p-value is 0.0433 and HR 8.673 (95% CI 1.067 to 70.520). Even this difference is therefore not statistically significant. When comparing the kinetics of marker levels, the difference is below statistical significance: p-value 0.6131 for P1NP and p-value 0.6357 for CTX. Conclusion: The results of this study confirm a correlation between the starting levels of CTX and P1NP with the overall survival rate, which corresponds to the other results presented in literature.

Predicting Vitality Change in Older Breast Cancer Survivors after Primary Treatment--an Approach Based on Using Time-related Difference of Pro-inflammatory Marker C-reactive Protein.

BACKROUND We aimed to determine prognosis of vitality change and functional status of breast cancer survivors after primary oncological treatment using time-related differences of elevated levels of highly sensitive proinflammatory C-reactive protein (CRP). PATIENTS AND METHODS The test group consisted of 46 elderly breast cancer survivors (median age was 65 years) who completed Vitality Scale of Short Form 36 (SF-36) after completing treatment and another retrospectively at diagnosis. Data on tumor-related factors, treatment, and outcomes were obtained retrospectively from medical records, and linear regression analysis was performed. CRP was followed at diagnosis and one year after primary treatment. Within the scope of this study, clinically important difference in the Vitality Scale was set at five points of change. RESULTS Results showed a statistically significant relationship between CRP change and vitality component of SF-36 change (rs = - 0.350, p = 0.023) in which a decrease in CRP inversely correlated with the quality of life component. The overall change was 1.078 of the vitality scale score (approximately 1 point) for each 1 unit decrease of CRP (1 mg/ L). Association of CRP levels (before and after treatment, its difference between these time points) with age, number of comorbidities and stage of the disease was analyzed and no statistically significant relationship was found in our study. CONCLUSION Preliminary results suggested time-related differences in elevated CRP levels as a potentially suitable predictor for change in vitality status for long term, chronic condition for older breast cancer survivors. We suggest the interpretation schema including an understanding that CRP change of 5 mg/ L and more should be considered a potential risk factor for subsequent negative clinical outcomes.