Multi-omic approach decodes paradoxes of the triple-negative breast cancer: lessons for predictive, preventive and personalised medicine.

Breast cancer epidemic in the early twenty-first century results in around two million new cases and half-a-million of the disease-related deaths registered annually worldwide. A particularly dramatic situation is attributed to some specific patient subgroups such as the triple-negative breast cancer (TNBC). TNBC is a particularly aggressive type of breast cancer lacking clear diagnostic approach and targeted therapies. Consequently, more than 50% of the TNBC patients die of the metastatic BC within the first 6 months of the diagnosis. In the current study we have hypothesised that multi-omic approach utilising blood samples may lead to discovery of a unique molecular signature of the TNBC subtype. The results achieved demonstrate, indeed, multi-omics as highly promising approach that could be of great clinical utility for development of predictive diagnosis, targeted prevention and treatments tailored to the person-overall advancing the management of the TNBC.

Breast cancer risk assessment: a non-invasive multiparametric approach to stratify patients by MMP-9 serum activity and RhoA expression patterns in circulating leucocytes.

Breast cancer is a multifactorial disease classified by several sub-types which differ from each other by risk factors, specific molecular promoters and severity of outcomes. Tumour aggressiveness and metastatic disease are the key determinants of breast cancer outcomes. Tumour cell ability to degrade the extracellular matrix and to be motile is the hallmark of invasion and essential step in a development of breast cancer metastatic disease. Therefore, a coordinated action between cell motility and ability to degrade the extracellular matrix is currently under extensive investigation focused on molecular targets for both diagnostic and therapeutic purposes. Contextually, our current study was dedicated to patient stratification utilising MMP-9 serum activity levels and RhoA expression patterns measured in circulating leucocytes. Biomarker patterns were "masked" in non-stratified patient groups. In contrast, the multiparametric stratification approach led to highly improved clinical utility of biomarker patterns. Presented stratification system is recommended for population screening as a cost-effective non-invasive approach to facilitate predictive diagnostics of breast cancer predisposition, pre-lesions and early stages, when the pathology can be effectively prevented or cured. Proposed approach might be particularly useful for early and predictive breast cancer diagnostics applied to certain phenotypes such as premenopausal rather than postmenopausal women, women with dense breast tissue, where highly increased RhoA/MMPs activities are utilised for effective proteolysis of the matrix and cancer cell migration into dense matrices, as well as for breast cancer of unclear origin such as particularly aggressive triple-negative sub-type.

Breast cancer epidemic in the early twenty-first century: evaluation of risk factors, cumulative questionnaires and recommendations for preventive measures.

Rapidly increasing incidence of breast cancer is a new social challenge resulting from a spectrum of internal and external risk factors which appear to be well accepted as an attribute of the early twenty-first century, being, however, new for female sub-populations compared to the past. These include altered socio-economical conditions such as occupational exposure, rotating shift work, specific environmental factors (increased pollution and environmental toxicity, altered dietary habits, quality and composition of meal) as well as consequently shifted and/or adapted physiologic factors such as lower age at menarche, late age of first full-term pregnancy, if any, shorter periods of breastfeeding and later menopause. Consolidated expert statements suggest that over 50 % of all breast cancer cases may be potentially prevented by risk reduction strategy such as regulation of modifiable risk factors. Currently available risk assessment models may estimate potential breast cancer predisposition, in general; however, they are not able to predict the disease manifestation individually. Further, current deficits in risk assessment and effective breast cancer prevention have been recently investigated and summarised as follows: gaps in risk estimation, preventive therapy, lifestyle prevention, understanding of the biology of breast cancer risk and implementation of known preventive measures. This paper overviews the most relevant risk factors, provides recommendations for improved risk assessment and proposes an extended questionnaire for effective preventive measures.

Patients with hepatic breast cancer metastases demonstrate highly specific profiles of matrix metalloproteinases MMP-2 and MMP-9 after SIRT treatment as compared to other primary and secondary liver tumours.

BACKGROUND: Patients with primary and metastatic liver malignancies represent a highly heterogeneous patient pool characterised by some of the shortest life expectancies amongst oncology patients. Investigation and better understanding of liver malignancies is an emerging field which requires high-quality multidisciplinary research and collaboration. METHODS: A study of 158 patients with primary hepatic carcinomas and secondary liver metastases, altogether 15 cancer types of different origin, who underwent selective internal radiation therapy (SIRT) with Yttrium(90) or transarterial chemoembolisation, was undertaken in an effort to detect distinguishing features with respect to activity profiles of both blood matrix metalloproteinase (MMP-2 and MMP-9). RESULTS: Noteworthy, stratification of all hepatic cancer groups with respect to MMP-2 and MMP-9 activities revealed characteristic patterns specifically in patients with hepatic breast cancer metastases who had undergone SIRT. In contrast to all other groups, these patients demonstrated well-consolidated profiles of both MMPs, reflecting a common feature, namely an immediate and durable increase of their activity after the SIRT treatment. Although the total number of patients in the breast cancer group is relatively small (15 patients), since increased activities of MMP-2 and MMP-9 are well known prognostic factors for poor outcomes of oncologic patients, the significance and clear group-specificity (from 15 ones investigated here) of this previously unanticipated finding requires particular attention and further investigations. Particularly important is to determine, whether this increase of the metalloproteinase activity was provoked by SIRT, as well as whether special selection criteria are required for patients with breast cancer metastases to the liver who are being considered for SIRT. CONCLUSIONS: It is recommended that a more focused, multidisciplinary and large-scaled investigations of the possible adverse effects of SIRT in patients with advanced metastatic disease of breast cancer be undertaken, with an appropriate patients' stratification, set-up of the relevant patient profiles and disease modelling.

Optimal multiparametric set-up modelled for best survival outcomes in palliative treatment of liver malignancies: unsupervised machine learning and 3 PM recommendations.

Over the last decade, a rapid rise in deaths due to liver disease has been observed especially amongst young people. Nowadays liver disease accounts for approximately 2 million deaths per year worldwide: 1 million due to complications of cirrhosis and 1 million due to viral hepatitis and hepatocellular carcinoma. Besides primary liver malignancies, almost all solid tumours are capable to spread metastases to the liver, in particular, gastrointestinal cancers, breast and genitourinary cancers, lung cancer, melanomas and sarcomas. A big portion of liver malignancies undergo palliative care. To this end, the paradigm of the palliative care in the liver cancer management is evolving from "just end of the life" care to careful evaluation of all aspects relevant for the survivorship. In the presented study, an evidence-based approach has been taken to target molecular pathways and subcellular components for modelling most optimal conditions with the longest survival rates for patients diagnosed with advanced liver malignancies who underwent palliative treatments. We developed an unsupervised machine learning (UML) approach to robustly identify patient subgroups based on estimated survival curves for each individual patient and each individual potential biomarker. UML using consensus hierarchical clustering of biomarker derived risk profiles resulted into 3 stable patient subgroups. There were no significant differences in age, gender, therapy, diagnosis or comorbidities across clusters. Survival times across clusters differed significantly. Furthermore, several of the biomarkers demonstrated highly significant pairwise differences between clusters after correction for multiple testing, namely, "comet assay" patterns of classes I, III, IV and expression rates of calgranulin A (S100), SOD2 and profilin-all measured ex vivo in circulating leucocytes. Considering worst, intermediate and best survival curves with regard to identified clusters and corresponding patterns of parameters measured, clear differences were found for "comet assay" and S100 expression patterns. In conclusion, multi-faceted cancer control within the palliative care of liver malignancies is crucial for improved disease outcomes including individualised patient profiling, predictive models and implementation of corresponding cost-effective risks mitigating measures detailed in the paper. The "proof-of-principle" model is presented.

Vasospastic individuals demonstrate significant similarity to glaucoma patients as revealed by gene expression profiling in circulating leukocytes.

PURPOSE: There is growing evidence that vasospatic individuals could be predisposed to develop glaucoma. Vasospastic deregulation is ensuing in activation of circulating leukocytes. In previous studies using "gene-hunting" strategies, we demonstrated stable alterations in gene expression profiles of circulating leukocytes isolated from glaucoma patients with vascular deregulation when compared to healthy individuals with no history of glaucomatous damage. The goal of this study was to look for possible similarities in gene expression profiles of circulating leukocytes in vasospastic individuals and glaucoma patients. METHODS: Normal-tension (NTG) and high-tension (HTG) glaucoma patients as well as individuals with vascular deregulation (VD) and healthy controls were recruited for the gene expression analysis. The methodology of comparative Expression Array analysis followed by highly sensitive quantitative real-time PCR has been used. RESULTS: Compared to the control group the expression of 146, 68, and 60 genes was found to be altered in NTG, HTG, and VD groups respectively. Thirty-four genes demonstrated similar expressional alterations in NTG, HTG, and VD groups versus controls, and only 21 genes demonstrated similar expressional alterations in NTG and HTG groups, having no overlap with the VD group. CONCLUSIONS: This result indicates a potential predisposition of vasospastic individuals to glaucomatous optic nerve atrophy. The targeted expression profiles might be further considered for early/predictive glaucoma diagnosis.

Down-regulation of microfilamental network-associated proteins in leukocytes of breast cancer patients: potential application to predictive diagnosis.

BACKGROUND: Breast cancer is one of the most frequent tumors worldwide. Currently applied diagnostic approaches are frequently not able to recognize early stages in tumor development therefore impairing outcomes. The focus of this study is the creation of a non-invasive predictive diagnostic approach by pathology-specific blood proteome analysis. PATIENTS AND METHODS: Circulating leukocytes were isolated from fresh blood samples of breast cancer patients, benign breast pathologies and healthy controls. In patients with all kinds of breast pathologies, blood samples were taken before core needle biopsy of the lump. Comparative protein mapping was performed by 2D-PAGE followed by MALDI-TOF analysis and Western-blot quantification of differentially expressed protein spots. RESULTS: By protein mapping, 64 protein spots were identified. Pathology-specific differential expression patterns comprised microfilamental network-associated proteins: Calgranulin A (S100), LyGDI (Rho GDIbeta), RhoA and profilin 1. RhoA and profilin values discriminated between healthy controls and patients with all breast pathologies. CONCLUSION: Microfilamental network-associated proteins are involved in the regulation of a variety of central cellular processes functionally linked with each other and known to be highly relevant for all stages of tumorigenesis including precancerous lesions and metastases. Pathology-related molecular patterns are currently considered for the creation of a novel highly sensitive minimally-invasive approach for predictive diagnosis of breast cancer.

Up-regulation of vimentin expression in low-density malignant glioma cells as immediate and late effects under irradiation and temozolomide treatment.

Nervous system tumors are one of the leading causes of cancer related death. Specific mechanisms facilitating the invasive behavior of gliomas remain obscure. Advanced simulation models of the in vivo response to therapy conditions should potentially improve malignant glioma treatment. Expressional profiling of vimentin--one of reliable pro-invasive tumor makers--in those simulation models was the goal of this study, in order to estimate a pro-invasive response of surviving malignant glioma cells under clinically relevant therapeutic conditions. Human U87-MG malignant glioma cells were used. These cells are characterized by the wild p53-phenotype, which is relevant for the majority of primary malignant glioblastomas. Experimental design foresaw the cells to undergo either irradiation or chemo-treatment with temozolomide alone, or combined treatment. Expression profiling of vimentin was performed by quantitative "Real-Time"-PCR under all treatment conditions simulating diverse tumor regions. Here we demonstrated that vimentin expression patterns in human malignant glioma cells strongly depend on cellular density, algorithms of drug delivery and chemo/radio treatment. Substantial differences were recognized between immediate and late therapy effects. Significant increase in vimentin expression levels was detected particularly in low-density cell cultures under durable treatment with constant concentration levels of temezolomide. Simulation of variable intratumoral regional conditions (central intratumoral regions vs. disseminated malignant cells in peripheral regions) demonstrated differential response of vimentin expression in malignant glioma cell cultures treated under clinically relevant conditions. Slight ebbing of expression levels as late effects of the treatment in confluent cultures may correspond to necrotic processes clinically observed in central intratumoral regions. Contrary, in disseminated malignant cells of peripheral regions therapy resulted in vimentin-inducing effects. This is in agreement with the clinical observations of an increased aggressiveness and malignancy grade of post-operatively chemo/radio-treated malignant gliomas.

Liquid biopsy and multiparametric analysis in management of liver malignancies: new concepts of the patient stratification and prognostic approach.

BACKGROUND: The annually recorded incidence of primary hepatic carcinomas has significantly increased over the past two decades accounting for over 800 thousand of annual deaths caused by hepatocellular carcinoma (HCC) alone globally. Further, secondary liver malignancies are much more widespread compared to primary hepatic carcinomas: almost all solid malignancies are able to metastasise into the liver. The primary tumours most frequently metastasising to the liver are breast followed by colorectal carcinomas. Given the increased incidence of both primary and metastatic liver cancers, a new, revised approach is needed to advance medical care based on predictive diagnostics, innovative screening programmes, targeted preventive measures, and patient stratification for treatment algorithms tailored to individualised patient profile. ADVANTAGES OF THE APPROACH TAKEN: The current pilot study took advantage of systemic alterations characteristic for liver malignancies, utilising liquid biopsy (blood samples) and specific biomarker patterns detected. Key molecular pathways relevant for pathomechanisms of liver cancers have been considered opening a perspective for both-individualised diagnostics and targeted treatment. Systemic alterations have been analysed prior to the therapy application avoiding molecular biological effects potentially diminishing predictive power of the biomarker-panel proposed. Multi-omics at DNA and protein (both expression and activity) levels has been applied. An established biomarker panel is considered as a powerful tool for individualised patient profiling and improved multi-level diagnostics-both predictive and prognostic ones. RESULTS AND CONCLUSIONS: Biomarker panels have been created for the patient stratification, prediction of a more optimal therapy and prognosis of survival based on the individualised patient profiling. Although there are some limitations of the pilot study performed, the results are encouraging, as it may be possible, through further research along these lines, to find a clinically and cost-effective means of stratifying liver cancer patients for personalised care and therapy. The benefits to the patient and society of accurate treatment stratification cannot be overemphasised.

Premenopausal breast cancer: potential clinical utility of a multi-omics based machine learning approach for patient stratification.

BACKGROUND: The breast cancer (BC) epidemic is a multifactorial disease attributed to the early twenty-first century: about two million of new cases and half a million deaths are registered annually worldwide. New trends are emerging now: on the one hand, with respect to the geographical BC prevalence and, on the other hand, with respect to the age distribution. Recent statistics demonstrate that young populations are getting more and more affected by BC in both Eastern and Western countries. Therefore, the old rule "the older the age, the higher the BC risk" is getting relativised now. Accumulated evidence shows that young premenopausal women deal with particularly unpredictable subtypes of BC such as triple-negative BC, have lower survival rates and respond less to conventional chemotherapy compared to the majority of postmenopausal BC. WORKING HYPOTHESIS: Here we hypothesised that a multi-level diagnostic approach may lead to the identification of a molecular signature highly specific for the premenopausal BC. A multi-omic approach using machine learning was considered as a potent tool for stratifying patients with benign breast alterations into well-defined risk groups, namely individuals at high versus low risk for breast cancer development. RESULTS AND CONCLUSIONS: The study resulted in identifying multi-omic signature specific for the premenopausal BC that can be used for stratifying patients with benign breast alterations. Our predictive model is capable of discriminating individually between high and low BC-risk with high confidence (>90%) and considered of potential clinical utility. Novel risk assessment approaches and advanced screening programmes-as the long-term target of this project-are of particular importance for predictive, preventive and personalised medicine as the medicine of the future, due to the expected health benefits for young subpopulations and the healthcare system as a whole.

Irradiated breast cancer patients demonstrate subgroup-specific regularities in protein expression patterns of circulating leukocytes.

BACKGROUND: Breast cancer is one of the most frequent types of cancer with fatal outcome worldwide. The use of breast conserving lumpectomy followed by radiation therapy is common and has been shown to be a strategy competitive to mastectomy in preventing mortality caused by breast cancer. However, breast irradiation, particularly applied after pre-irradiation chemotherapy, frequently leads to serious short- and long-term side-effects, the prediction of which is highly desirable in terms of individual therapy planning. For these purposes, minimal-invasive molecular blood analysis is considered as a powerful diagnostic tool: molecular interplay in blood is highly informative and may predict individual side-effects of therapy. MATERIALS AND METHODS: Ex vivo comparative protein expression profiling was performed in circulating leukocytes isolated from fresh blood samples of seven breast cancer patients before lumpectomy and consequently at several checkpoints under radiation treatment (0-60 Gy). Protein expression patterns were investigated by two-dimensional polyacrylamide gel electrophoresis followed by protein spot identification using matrix assisted laser desorption/ ionisation -- time of flight. Specific expression levels of highly affected differentially expressed proteins were quantified by Western blotting. RESULTS: The radiation treatment caused individual extensive alterations in expression patterns of leukocytes in the patients tested. In particular, a key regulator of redox status, thioredoxin, and the free-radical detoxification cascade members, SOD-2 and catalase, were highly affected. In spite of the high diversity of individual expression levels, characteristic protein expression patterns were recognized and patients were grouped according to the similarities found. CONCLUSION: Characteristic expression patterns in circulating leukocytes might provide novel molecular targets for prediction of therapy side-effects and improve individual therapy planning for breast cancer patients, thus avoiding unnecessary and excessive treatment-related toxicity. Molecular candidates and specific patterns are demonstrated in this work.

Impaired wound healing: facts and hypotheses for multi-professional considerations in predictive, preventive and personalised medicine.

Whereas the physiologic wound healing (WH) successfully proceeds through the clearly defined sequence of the individual phases of wound healing, chronic non-healing wounds/ulcers fail to complete the individual stages and the entire healing process. There are many risk factors both modifiable (such as stress, smoking, inappropriate alcohol consumption, malnutrition, obesity, diabetes, cardio-vascular disease, etc.) and non-modifiable (such as genetic diseases and ageing) strongly contributing to the impaired WH. Current statistics demonstrate that both categories are increasingly presented in the populations, which causes dramatic socio-economic burden to the healthcare sector and society at large. Consequently, innovative concepts by predictive, preventive and personalised medicine are crucial to be implemented in the area. Individual risk factors, causality, functional interrelationships, molecular signature, predictive diagnosis, and primary and secondary prevention are thoroughly analysed followed by the expert recommendations in this paper.

Pro-invasive gene regulating effect of irradiation and combined temozolomide-radiation treatment on surviving human malignant glioma cells.

The current chemotherapeutic treatment of glioblastoma patients has minor success. Little is known about the molecular and cellular mechanisms of the resistance of gliomas towards current therapies. This study investigated both suppressive cellular effects and regulation of extracellular matrix remodeling proteins with pro-invasive activity in surviving human glioblastoma cells under clinically relevant treatments. All cellular and molecular biological investigations were performed on the genetically well-defined and clinically relevant p53-wild type U87Mg glioma cells. Malignant glioma cells underwent either radiation or temozolomide treatments alone, or combined chemo/radio treatment. Protein expression patterns were investigated by two-dimensional polyacrylamide gel electrophoresis followed by protein spot identification using tandem mass spectrometry analysis. Specific expression levels were quantified by Western-blotting. Extracellular gelatinase activities for both metalloproteinases MMP-2 and MMP-9 were determined by zymogramms. Survival curves indicated no effective suppression of glioma cells under all treatment conditions tested. Morphological changes demonstrated sub-lethal effect of both temozolomide and combined treatment. Expression of MMP-2, MMP-9, and membrane type 1 matrix metalloproteinases (MT1-MMP) was differentially up-regulated by increasing cellular density and treatment conditions. A significantly enhanced extracellular degrading activity under all treatment conditions tested was demonstrated for MMP-2 only. Being a marker for brain tumour progression and angiogenesis, lysozyme c was highly up-regulated under the combined chemo/radio treatment. The activation of proteins with pro-invasive activity indicates an increasing malignancy grade of surviving glioma cells under treatment conditions tested correlating well with more aggressive tumour phenotypes observed clinically in recurrences of treated glioblastomas.

Potential biomarker panels in overall breast cancer management: advancements by multilevel diagnostics.

Breast cancer (BC) prevalence has reached an epidemic scale with half a million deaths annually. Current deficits in BC management include predictive and preventive approaches, optimized screening programs, individualized patient profiling, highly sensitive detection technologies for more precise diagnostics and therapy monitoring, individualized prediction and effective treatment of BC metastatic disease. To advance BC management, paradigm shift from delayed to predictive, preventive and personalized medical services is essential. Corresponding step forwards requires innovative multilevel diagnostics procuring specific panels of validated biomarkers. Here, we discuss current instrumental advancements including genomics, proteomics, epigenetics, miRNA, metabolomics, circulating tumor cells and cancer stem cells with a focus on biomarker discovery and multilevel diagnostic panels. A list of the recommended biomarker candidates is provided.

Selective internal radiation therapy in treatment of hepatocellular carcinoma: new concepts of personalization.

Hepatocellular carcinoma (HCC) is a global health problem, with more than half a million new cases diagnosed annually and mortality rates at similar level. The majority of HCC is diagnosed at intermediate-advanced stages being, therefore, an issue for palliative rather than curative care. Selective internal radiation therapy (SIRT) is one of the best appropriate palliative treatment modalities in HCC management. Although delivering satisfactory results, SIRT application comes along with frequent complications and tumor recurrence. Recent studies suggest treatment algorithm tailored to the person as improving individual outcomes and reducing treatment-related complications. This review provides insights to implicate innovative concepts of predictive, preventive and personalized medicine in SIRT application to HCC cohorts.

Persistence of Chlamydia pneumoniae in degenerative aortic valve stenosis indicated by heat shock protein 60 homologues.

BACKGROUND AND AIMS OF THE STUDY: Based on the concept of chronic persistent infections with Chlamydia pneumoniae among variable stressors for aortic valve degeneration, the study aim was to assess the presence of chlamydial heat shock protein (cHSP) 60 and its human homologue (hHSP60) in diseased valvular tissue. METHODS: Surgical specimens of high-grade stenosed, native (n = 33) and bioprosthetic (n = 10) aortic valves were examined immunohistochemically for the localization of cHSP60, hHSP60 and macrophages (CD68), supplemented by polymerase chain reaction (PCR) and electron microscopy to prove microbial presence. RESULTS: Degenerated valves showed specific immunostaining of cHSP60 in 27 cases (65%), of hHSP60 in 26 (63%), and of CD68 in 36 (84%). Both HSP60 homologues were predominantly detected in valvular fibrosa, consistently co-localized with macrophages and, quantitatively, showed a strong correlation (r = 0.81, p < 0.001). Presence of C. pneumoniae was demonstrated by PCR in a subset of 11 of 18 valves (61%). Microbial persistence was confirmed by ultrastructural analysis. Degenerated prosthetic valves revealed markedly higher macrophage infiltration and cHSP60 signaling compared with degenerated native valves (each p < 0.05). CONCLUSION: Beyond detection of C. pneumoniae, the present data on co-localization and valvular predilection sites (fibrosa) of both HSP60 homologues indicate the presence of chronic persistent C. pneumoniae infection as well as regional stressor effects, and suggest their involvement in native and prosthetic valve degeneration.

Increased expression of matrix metalloproteinases in mononuclear blood cells of normal-tension glaucoma patients.

PURPOSE: Glaucomatous optic neuropathy involves tissue remodeling by matrix metalloproteinases (MMPs). In this study we investigated MMP gene expression in circulating leukocytes isolated from normal tension glaucoma (NTG) patients in comparison to healthy controls. METHODS: Blood samples were collected from 6 glaucoma patients and 6 age- and sex-matched healthy controls. Leukocytes were separated using Ficoll-Hypaque gradient. mRNA pools were used for subtractive hybridization to identify genes with altered expression. The subtracted genes were sequenced and individual mRNA pools were quantified using semiquantitative RT-PCRs. Target PCR products were confirmed using sequence-based restriction analysis. In this study we focused on MMPs. RESULTS: MMP-9 and MT1-MMP (MMP-14) were subtracted as upregulated genes in the group of NTG patients. Upregulation of these genes was confirmed by RT-PCR and Western-blot analysis in all 6 patients. The expression of the tissue inhibitor of matrix metalloproteinases TIMP-1 was slightly enhanced in patients as compared with controls. Expression of MMP-2 was not detected in leukocytes either in glaucoma patients or in healthy subjects. CONCLUSION: A simultaneous upregulation of both MMP-9 and MT1-MMP gene expression and only slightly enhanced expression of TIMP-1 suggest an increased enzymatic matrix metalloproteinase activity delivered by mononuclear blood cells in these patients.

Differences in gene expression in lymphocytes of patients with high-tension, PEX, and normal-tension glaucoma and in healthy subjects.

PURPOSE: Purpose Differences in the gene expression of leukocytes between patients with normal-tension glaucoma (NTG) and controls have been described. This study was performed in order to detect the differences in gene expression in peripheral lymphocytes in patients with primary open-angle glaucoma (POAG), patients with pseudoexfoliation glaucoma (PEX), and patients with NTG, and in healthy subjects. METHODS: Ten patients with POAG, 11 patients with PEX, 10 patients with NTG, and 42 sex- and age-matched healthy persons were recruited. All study subjects were Caucasian. Twenty-two preselected genes were chosen and their expression in blood lymphocytes was quantified by real-time PCR. First, a univariate comparison among all groups was performed using the nonparametric Friedman test. Second, an L1 penalized logistic regression was performed. RESULTS: Using the Friedman test to compare the 4 groups, 9 genes showed a different expression (p<0.05). Comparing the controls vs patients with POAG, 8 genes were differently expressed (p<0.05). Comparing patients with PEX vs controls, 9 genes were significantly different (p≤0.05). The statistical analysis of patients with NTG vs controls showed a difference in gene expression of 7 genes (p≤0.05). All these genes were upregulated in the glaucoma groups compared with the controls. The genes RhoGDI and RAR showed the most significant statistical difference in the L1-penalized logistic regression. The genes overexpressed in POAG/PEX differed from the ones in NTG. CONCLUSIONS: In this masked study among the preselected 22 genes, several genes are overexpressed in the blood lymphocytes of Caucasian patients with glaucoma compared with the controls. The genes upregulated in POAG/PEX differed from the ones in NTG.

'Suspect molecular signature' in blood as the indicator of undiagnosed breast cancer, cancer risk and targeted prevention.

BACKGROUND: Breast cancer is a multifactorial disease with the highest incidence rates amongst all cancer types. Further, high levels of circulating tumour cells are a characteristic of breast cancer patients demonstrating a particular predisposition to the development of breast cancer metastatic disease. Actual diagnostic approaches are frequently unable to recognise early stages of tumour development which impairs individual outcomes. In contrast, predictive and preventive risk assessment and early diagnosis may lead to full recovery after surgical resection. Recently, the authors have reported about the construction of diagnostic windows, which could influence the molecular diagnostics of breast cancer. MATERIAL AND METHODS: In a previous study, diagnostic windows for breast cancer risk assessment were analysed. Women with non-malignant breast diseases demonstrating molecular profiles similar to those of breast cancer patients were enrolled into this follow-up study. In the interviews, for patients identified as predisposed to cancer, a specialised questionnaire has been set up to characterise individual risk factors and estimate their potential impacts on cancer onset and progression. RESULTS AND CONCLUSIONS: By utilising the technological tool of diagnostic windows, 13 individuals have been identified demonstrating molecular profiles typical for patients diagnosed with breast cancer. The current paper summarises the analytical results and makes statements to the application of the pathology-specific molecular profiles recognised as the technological tool for improved diagnostic approach, breast cancer risk assessment and preventive health care management. The necessity to create individual patient profiles and analyse the evolution of the molecular signature is justified for advanced medical services. Expert recommendations are provided to promote further developments in the field of advanced breast cancer management.

Risk assessment, disease prevention and personalised treatments in breast cancer: is clinically qualified integrative approach in the horizon?

Breast cancer is a multifactorial disease. A spectrum of internal and external factors contributes to the disease promotion such as a genetic predisposition, chronic inflammatory processes, exposure to toxic compounds, abundant stress factors, a shift-worker job, etc. The cumulative effects lead to high incidence of breast cancer in populations worldwide. Breast cancer in the USA is currently registered with the highest incidence rates amongst all cancer related patient cohorts. Currently applied diagnostic approaches are frequently unable to recognise early stages in tumour development that impairs individual outcomes. Early diagnosis has been demonstrated to be highly beneficial for significantly enhanced therapy efficacy and possibly full recovery. Actual paper shows that the elaboration of an integrative diagnostic approach combining several levels of examinations creates a robust platform for the reliable risk assessment, targeted preventive measures and more effective treatments tailored to the person in the overall task of breast cancer management. The levels of examinations are proposed, and innovative technological approaches are described in the paper. The absolute necessity to create individual patient profiles and extended medical records is justified for the utilising by routine medical services. Expert recommendations are provided to promote further developments in the field.