Prognostic and Treatment Predictive Markers Associated with Cyclin D1 Gene Amplification and Epithelial-Mesenchymal Transition in Breast Cancer

Endocrine resistant breast cancer is a major therapeutical challenge and accurate selection of patients susceptible to certain types of anti-hormonal treatment strategies is pivotal. Discovery of novel biomarkers potentially predicting treatment response as well as the clinical course of breast cancer is essential to effectively improve therapies and diagnosis. The main goal of this thesis was to identify putative markers for prediction of clinical outcome and response to tamoxifen, the first-line endocrine treatment option for premenopausal breast cancer patients. The markers examined were associated with CCND1 amplification (Chk1 and β-arrestin1), which has previously been linked to a potential adverse effect of tamoxifen, or with epithelial-mesenchymal transition (Snail), a biological process crucial for the development of metastasis. Immunohistochemical analyses of the biomarkers were predominantly performed in a breast cancer cohort including premenopausal patients randomly assigned to receive either tamoxifen or no adjuvant treatment. The in vitro studies investigated the regulation of EMT in hypoxia, in a panel of human breast cancer cells lines. We found that deletion of the CHK1 gene is potentially associated with CCND1 amplification, and loss of Chk1 protein expression was demonstrated to be linked to an impaired response to tamoxifen in premenopausal breast cancer patients. Additionally, the EMT-regulator Snail, which has been reported to be involved in ER- signaling, was shown to predict tamoxifen response, where absence of nuclear Snail rendered patients less sensitive to this adjuvant therapy. Moreover, hypoxia induced an incomplete EMT in vitro, and Snail modulated the migratory propensity of breast cancer cells. Finally, stromal expression of β-arrestin1 was found to be an independent prognostic marker in a cohort of both pre- and postmenopausal breast cancer patients. In premenopausal patients both high expression and absence of β-arrestin1 was linked to poor prognosis, but expression of this protein was not associated with an altered tamoxifen response. In conclusion, these studies revealed two biomarkers with potential clinical significance in predicting the sensitivity to adjuvant tamoxifen, as well as one with an impact on clinical outcome

A Triple-Transgenic Immunotolerant Mouse Model

ABSTRACTAvoiding unwanted immunogenicity is of key importance in the development of therapeutic drug proteins. Animal models are of less predictive value because most of the drug proteins are recognized as foreign proteins. However, different methods have been developed to obtain immunotolerant animal models. So far, the immunotolerant animal models have been developed to assess one protein at a time and are not suitable for the assessment of combination products. Our aim was to develop an animal model for evaluating the impact of manufacturing and formulation changes on immunogenicity, suitable for both single protein and combination products. We constructed two lines of transgenic mice expressing the three human coagulation factors, II, VII, and X, by inserting a single vector containing the three coagulation factors encoding sequences separated by insulator sequences derived from the chicken beta-globin locus into the mouse genome. Immunization of transgenic mice from the two lines and their wild-type littermates showed that transgenic mice from both lines were immunotolerant to the expressed human coagulation factors. We conclude that transgenic mice immunotolerant to multiple proteins can be obtained, and that these mice are potentially useful as animal models in the assessment of immunogenicity in response to manufacturing changes. © 2013 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 102:1116-1124, 201

Gene products of chromosome 11q and their association with CCND1 gene amplification and tamoxifen resistance in premenopausal breast cancer

Introduction: The amplification event occurring at chromosome locus 11q13, reported in several different cancers, includes a number of potential oncogenes. We have previously reported amplification of one such oncogene, namely CCND1, to be correlated with an adverse effect of tamoxifen in premenopausal breast cancer patients. Over-expression of cyclin D-1 protein, however, confers tamoxifen resistance but not a tamoxifen-induced adverse effect. Potentially, co-amplification of an additional 11q13 gene, with a resulting protein over-expression, is required to cause an agonistic effect. Moreover, during 11q13 amplification a deletion of the distal 11q region has been described. In order to assess the potential impact of the deletion we examined a selected marker for this event. Method: Array comparative genomic hybridization analysis was employed to identify and confirm changes in the gene expression of a number of different genes mapping to the 11q chromosomal region, associated with CCND1 amplification. The subsequent protein expression of these candidate genes was then examined in a clinical material of 500 primary breast cancers from premenopausal patients who were randomly assigned to either tamoxifen or no adjuvant treatment. The protein expression was also compared with gene expression data in a subset of 56 breast cancer samples. Results: Cortactin and FADD (Fas-associated death domain) over-expression was linked to CCND1 amplification, determined by fluorescence in situ hybridization, but was not associated with a diminished effect of tamoxifen. However, deletion of distal chromosome 11q, defined as downregulation of the marker Chk1 (checkpoint kinase 1), was associated with an impaired tamoxifen response, and interestingly with low proliferative breast cancer of low grade. For Pak1 (p21-activated kinase 1) and cyclin D-1 the protein expression corresponded to the gene expression data. Conclusions: The results indicate that many 11q13 associated gene products are over-expressed in conjunction with cyclin D-1 but not linked to an agonistic effect of tamoxifen. Finally, the deletion of distal 11q, linked to 11q13 amplification, might be an important event affecting breast cancer outcome and tamoxifen response

The Shepherds' Tale : A Genome-Wide Study across 9 Dog Breeds Implicates Two Loci in the Regulation of Fructosamine Serum Concentration in Belgian Shepherds

Diabetes mellitus is a serious health problem in both dogs and humans. Certain dog breeds show high prevalence of the disease, whereas other breeds are at low risk. Fructosamine and glycated haemoglobin (HbA1c) are two major biomarkers of glycaemia, where serum concentrations reflect glucose turnover over the past few weeks to months. In this study, we searched for genetic factors influencing variation in serum fructosamine concentration in healthy dogs using data from nine dog breeds. Considering all breeds together, we did not find any genome-wide significant associations to fructosamine serum concentration. However, by performing breed-specific analyses we revealed an association on chromosome 3 (rho(corrected) approximate to 1:68 x 10(-6)) in Belgian shepherd dogs of the Malinois subtype. The associated region and its close neighbourhood harbours interesting candidate genes such as LETM1 and GAPDH that are important in glucose metabolism and have previously been implicated in the aetiology of diabetes mellitus. To further explore the genetics of this breed specificity, we screened the genome for reduced heterozygosity stretches private to the Belgian shepherd breed. This revealed a region with reduced heterozygosity that shows a statistically significant interaction (rho = 0.025) with the association region on chromosome 3. This region also harbours some interesting candidate genes and regulatory regions but the exact mechanisms underlying the interaction are still unknown. Nevertheless, this finding provides a plausible explanation for breed-specific genetic effects for complex traits in dogs. Shepherd breeds are at low risk of developing diabetes mellitus. The findings in Belgian shepherds could be connected to a protective mechanism against the disease. Further insight into the regulation of glucose metabolism could improve diagnostic and therapeutic methods for diabetes mellitus.Peer reviewe

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Viktimisering bland familjehemsplacerade ungdomar i Stockholm: förekomst och förändring över tid

Prior studies have shown that individuals with experiences of child welfare interventions have high rates of adverse social and health outcomes. Yet little is known about whether they are disproportionately subjected to crime victimization. Using repeated cross-sectional data for 32,332 adolescents in Stockholm (of which around 1% is placed in family foster care), this study asks whether adolescents placed in family foster care have higher rates of crime victimization compared to same aged peers living in other family types. The aim is also to examine if this trend is the same for adolescents placed in family foster care over time.Results from gender stratified logistic regression analyses show that adolescents placed in family foster care have a higher risk of reporting various forms of crime victimization compared to adolescents living in intact families. The differences were less pronounced when compared to adolescents living in single parent families. With the exception of having experienced severe threats, where they show an increase in risk over time, results from interaction analyses suggest that adolescents in family foster care generally follow the same decreasing trend as their peers living in other family types. Implications for research, policy and practice are discussed.

Hypoxia and breast cancer: prognostic and therapeutic implications

Hypoxia affects many important processes in tumour progression and is a key feature in the tumour microenvironment that needs to be taken into account when evaluating prognostics and therapeutic options for cancer patients. Hypoxia-regulating proteins, i.e. hypoxia inducible factors (HIFs), and associated gene products have been linked to certain tumour behaviours and might be useful as prognostic and predictive markers. Recently, hypoxia-driven gene products have been launched as novel cancer treatment targets with the potential to increase tumour-specific effects. Breast cancer consists of a multitude of different diseases with certain common characteristics, but also clearly disparate behaviours and genetic alterations. In this review we will summarise the role of hypoxia in breast cancer and specifically outline the importance of hypoxia and HIF-1alpha regarding prognostic and treatment-specific implications

Cultural policy in the Nordic welfare states: aims and functions of public funding for culture

In this research anthology on public subsidy systems for culture in the Nordic region, researchers from each Nordic country contribute with a chapter on the status and challenges of public subsidy systems for culture in their particular country. In addition, a former civil servant with the Nordic Council of Ministers provides descriptions of Nordic co-operation grants for culture, as well as grants in the Faroe Islands, Greenland, and Åland. While the authors have chosen which issues to focus on in their respective chapters, all in one way or another concern themselves with the question of how Nordic welfare policies are reflected in Nordic cultural policies. The research anthology has been produced by Kulturanalys Norden and edited by Sakarias Sokka, senior researcher at CUPORE