Дисертації з теми "Lung and breast"

The objective of the present study is to investigate various problems associate with breast cancer and lung cancer patients. In this study, we compare the effectiveness of breast cancer treatments using decision tree analysis and come to the conclusion that although certain treatment shows overall effectiveness over the others, physicians or doctors should discretionally give different treatment to breast cancer patients based on their characteristics. Reoccurrence time of breast caner patients who receive different treatments are compared in an overall sense, histology type is also taken into consideration. To further understand the relation between relapse time and other variables, statistical models are applied to identify the attribute variables and predict the relapse time. Of equal importance, the transition between different breast cancer stages are analyzed through Markov Chain which not only gives the transition probability between stages for specific treatment but also provide guidance on breast cancer treatment based on stating information. Sensitivity analysis is conducted on breast cancer doubling time which involves two commonly used assumptions: spherical tumor and exponential growth of tumor and the analysis reveals that variation from those assumptions could cause very different statistical behavior of breast cancer doubling time. In lung cancer study, we investigate the mortality time of lung cancer patients from several different perspectives: gender, cigarettes per day and duration of smoking. Statistical model is also used to predict the mortality time of lung cancer patients.

Breast Cancer (BC) is one of the major causes of cancer deaths in women. BC is a heterogeneous disease, and within the heterogeneity of the tumor, Tumor-Initiating Cells (TICs) have been reported as important drivers in tumor initiation and progression. On the basis of these observations, we hypothesized that TICs contribute to BC metastasis, as well to chemoresistance. Through genetic, transcriptomic, molecular and therapeutic analyses of tumor xenografts, BC cell lines, and/or human tumors, in this thesis we first demonstrate that RARRES3, a lung metastatic suppressor gene, prevents adhesion to the lung parenchyma and the initiation of metastatic lesions by enforcing the retention of differentiation features and that this retention is driven by its PLA1/2 catalytic activity. These results indicate that RARRES3 genetic activity blocks both tissue-specific metastasis to the lung and metastatic initiation properties. Next, we dissected functional interplay between stem cell (SC)-like master regulator genes (EVI1 and SOX9) and resistance to mTOR inhibitors, which leads to an aggressive metastatic cancer phenotype. Collectively, the data shown in this thesis depict novel evidence regarding the role of tumor initiation properties in: I) BC progression and metastasis; II) how cancer progression is functionally linked to resistance to mTOR; and III) how resistance to therapy driven by tumor initiation properties may eventually produce an aggressive cancer phenotype.
El cáncer de mama (BC, de sus siglas en inglés) es una de las mayores causas de muerte por cáncer en mujeres. El cáncer de mama es una enfermedad heterogénea, y en la heterogeneidad del tumor, las células iniciadoras tumorales (TICs, del inglés) han sido asociadas como importantes responsables en la iniciación y progresión tumoral. Basándonos en estas observaciones, nuestra hipótesis es si estas células contribuyen en la metástasis y quimioresistancia en el cáncer de mama. A través de análisis genéticos, transcriptómicos, moleculares y terapéuticos en xenoinjertos tumorales, líneas celulares y tumores humanos, en esta tesis hemos revelado en primer lugar que RARRES3, es un gen de supresión metastática en pulmón, que previene la adhesión al parénquima pulmonar y la iniciación de lesiones metastáticas, imponiendo las características de diferenciación, y cuya retención es causada por la actividad catalítica PLA1/2. Estos resultados indican que la actividad genética de RARRES3 bloquea específicamente la metástasis a pulmón y las propiedades de iniciación metastática. Además, hemos descrito una relación entre las los genes con características de autorenovación (EVI1 y SOX9) y resistencia a inhibidores de mTOR, que termina con una fenotipo más agresivo y metastático. Conjuntamente, los datos mostrados en esta tesis demuestras evidencias novedosas relacionadas con las propiedades de iniciación tumoral en distintos contextos: I) progresión y metástasis en el cáncer de mama; II) como la progresión tumoral está funcionalmente relacionada con la resistencia a inhibición de mTOR; y III) como la resistencia a terapia que está desencadenada por estas propiedades de iniciación tumoral pueden al final producir un fenotipo más agresivo y metastático

RARbeta2 is a tumour suppressor activated by retinoic acid (RA). It controls gene expression by heterodimerizing with RXRs on RA response elements. We hypothesised that Erk1/2 inhibits RARbeta2 transcriptional activity in RARbeta2-expressing and RA-resistant cell lines by phosphorylating RXRalpha. Treatment of RA-resistant, RARbeta2-expressing cell lines (A549 and Hs578t) and a RA sensitive cell line (H157) with PD98059 synergized with RA in inhibiting growth. Cloning an active mutant of MEK1 (MEK1DD) into H157 made these cells around 40% less sensitive to ATRA. EGF-mediated Erk1/2 activation inhibited the ATRA-induced transcription of both a synthetic reporter and an endogenous (RARbeta) gene. In both cases, PD98059 reversed this inhibition. Finally, MEK1DD inhibited to approximately 90% transcription from the reporter gene. These results indicate that Erk1/2 reduces RA-mediated transcription by all RARs, possibly by phosphorylating RXRalpha and that RA may be effective clinically when used in combination with drugs acting upstream of MEK1/2 such as Iressa and Herceptin.

The purpose of this thesis is to explore the theory, practice and application of costing with specific reference to cancer. In part it reviews the theory and guidelines related to costing methods including the recent focus on the analytical techniques used with cost data. In addition it examines how these theories and guidelines are applied in practice, by reviewing the literature on costs and cancer. The empirical research in this thesis applies costing methods to three specific cancer sites; breast, cervix and lung. This analysis provides information on the total burden of these specified cancers in terms of cost to a typical health authority (Trent). It also explores the hypothesis highlighted in previous studies that the cost of cancer increases with the stage of the disease. The final area of contribution for the thesis is in the application of recently suggested analytical techniques for cost data to the breast, cervical and lung cancer data sets; it investigates a number of proposed techniques for the analysis of skewed cost data and methods for data with incomplete patient follow up.

Radiotherapy cures many cancers, but the optimum total doses and fractionations used to treat different cancer types remain uncertain. While conventional fractionation (≈2 Gy per fraction) is common in many countries, UK practice has been highly variable. This thesis compared different curative-intent radiotherapy dose-fractionations used in non-small cell lung and breast cancer. These two cancers together make up over a quarter of UK cancer incidence and mortality, and radiotherapy can increase cure rates of both cancers. Two studies were conducted: (A) A meta-analysis of randomised radiotherapy trials in non-small cell lung cancer and (B) A cohort study of non-small cell lung and breast cancer radiotherapy in the Thames Valley. For the meta-analysis, a systematic search was conducted. Eligible studies were randomised comparisons of two or more radiotherapy regimens. Median survival ratios were calculated for each comparison and pooled. 3,795 patients in 25 randomised comparisons of radiotherapy dose were studied. When radiotherapy was given alone, the higher dose within-trial resulted in increased survival (median survival ratio 1.13, 95% confidence interval 1.04-1.22). When radiotherapy was given with concurrent chemotherapy, the higher dose within-trial resulted in decreased survival (median survival ratio 0.83, 95% confidence interval 0.71-0.97). For the cohort study, multiple Public Health England data sources were combined to obtain information on radiotherapy, patient characteristics, and outcomes. Multivariable Cox regressions were conducted separately by cancer site. 324 non-small cell lung, 8,879 invasive breast, and 477 ductal carcinoma in situ patients were studied. In analyses of both non-small cell lung and invasive breast cancer, increasing radiotherapy dose was associated with improved survival in some treatment centres, while in other centres the opposite was true. These opposite trends by treatment centre were unlikely to be explained by chance, and they suggest that differences in patient selection were driving results. There were insufficient events among ductal carcinoma in situ patients to assess associations. Findings from the meta-analysis support consideration of further radiotherapy dose escalation trials, making use of modern methods to reduce toxicity. Findings from the cohort study suggest that it is not possible to use observational studies to examine causal effects of radiotherapy dose-fractionation. This thesis therefore shows the continued importance of conducting sufficiently large randomised trials to ascertain optimal dose-fractionation in radiotherapy.

Cancer is the second biggest cause of death in the economically developed countries with breast cancer in females and lung cancer in males are the most frequently diagnosed tumours. Clinically referred to as malignant neoplasias, cancer is a combination of patho- logical states, all involving unregulated proliferation of cells in speficic locations. Over the last few years a new concept has emerged, suggesting that cancer is a disease driven by breast tumour-initiating cells (BTICs). Most current anti-cancer therapies used in clinical practice have the capability to reduce the tumour size, however they have detrimental side effects and also fail to target BTICs, a phenomenon that often leads to recurrence of tumours and disease relapse. The novel anti-cancer agents, referred to as mitocans are often selective for cancer cells while being non-toxic to normal cells and may have the propensity to induce apoptosis in tumour-initiator cells including BTICs. Our group has focused on two mitocans, -tocopheryl succinate and its mitochondrially targeted derivative MitoVES. While it has been documented that MitoVES is effective in a variety of cancer cells and in vivo mouse models of cancer, it has not been established what effects it has on BTICs and whether it acts by aiming at relevant molecular targets. The Thesis addresses these questions by establishing three aims, which are addressed in individual chapters. We have validated that the breast cancer cell line MCF7-derived mammospheres (MS) is a valid model to study the BTIC biology and resistance to apoptosis, as both phenotypical and genotypical features confirm increased level of stemness.
Thesis (PhD Doctorate)
Doctor of Philosophy (PhD)
School of Medical Science
Griffith Health
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Fibroblasts are abundant mesenchymal cells present in all tissues in a quiescent state, which contribute to wound healing when activated. Cytokine transforming growth factor-β1 (TGF-β1) stimulates fibroblast-myofibroblast differentiation, which induces extracellular matrix secretion, tissue contraction and promotes cancer cell migration. Hence, chronic activity of stromal myofibroblasts correlates with a poor prognosis for cancer and organ fibrosis patients. Therefore, modulating myofibroblast activity may reduce the severity of these diseases. Previous research suggests blockade of transmembrane integrin receptors expressed by fibroblasts prevents TGF-β1- induced differentiation, indicating integrins are attractive therapeutic targets. However, fibroblasts derived from different organs exhibit heterogeneity, although their integrin expression and integrin-regulated differentiation has not been directly compared. The aim of my research was 1) to understand and compare how integrins regulate TGF-β1-induced activation of fibroblasts derived from normal skin, lung and breast tissue; 2) to examine the global gene expression of TGF-β1-treated lung fibroblasts; 3) to identify novel therapeutic targets that modulate TGF-β1-induced activation of lung fibroblasts using a drug library. qPCR showed skin, lung and breast fibroblasts differentially expressed TGF-β1- induced activation markers, including ACTA2, FN1, TIMP3, CTGF and SERPINE1, in addition to integrin genes for α1, α4, α11 and β3. Small-molecule inhibitors of αv integrins only reduced the invasion of TGF-β1-exposed skin fibroblasts, but not lung or breast fibroblasts. siRNA against α11, β3 and β5 decreased TGF-β1-induced collagen contraction and activation marker expression in skin and lung fibroblasts, while α1 siRNA prevented collagen contraction by breast fibroblasts only. RNA sequencing of TGF-β1-treated lung fibroblasts revealed pro-inflammatory and profibrotic pathways were significantly enriched, while screening TGF-β1-treated lung fibroblasts with a FDA-approved drug library identified 46 hits that significantly reduced α-smooth muscle actin and fibronectin expression. Overall, genes are differentially expressed in TGF-β1-treated skin, lung and breast fibroblasts, while different integrins in each fibroblast appear to regulate invasion, TGF-β1-induced collagen contraction and gene expression. RNA sequencing revealed TGF-β1 promotes the expression of a pro-tumour signature in lung fibroblasts and several novel therapeutic targets that modulate the activation of lung fibroblasts have been identified. Understanding these integrin-dependent and independent mechanisms will facilitate the generation of myofibroblast-targeted treatments for cancer and organ fibrosis.

In order to study the expression of MMP2, MMP 3 and MMP9 in breast cancer brain and lung metastasis, we used a syngeneic rat model of distant metastasis of ENU1564, a carcinogen-induced mammary adenocarcinoma cell line. At six weeks post inoculation we observed development of micro-metastasis in the brain and lung. Immunohistochemistry and Western blotting analyses showed that MMP 2, -3 and -9 protein expression is consistently significantly higher in neoplastic brain tissue compared to normal brain tissue. Lung metastases express abundant MMP2, -3 and -9 in neoplastic cell cytoplasm. In situ zymography revealed gelatinase activity within the brain metastasis. Gel zymography showed an increase in MMP2 and MMP3 activity in brain metastasis. Furthermore, we were able to significantly decrease the development of breast cancer brain and lung metastasis in animals by treatment with PD 166793, a selective synthetic MMP inhibitor. In addition, PD 166793 decreased the in vitro invasive cell behavior of ENU1546. TIMP2 overexpression also decreased the development of breast cancer lung metastasis in our model. Our results suggest that MMP2, -3 and -9 may be involved in the process of metastasis of breast cancer to the brain and lung. Because astrocytes have been associated with breast cancer brain metastasis we evaluated the role of astrocytes and ERK2 pathway in MMP2 up-regulation in BC brain metastasis. A significant decrease in brain metastases development, and orthotopic tumor size and weight were observed in animals inoculated with ENU1564-TIMP2 cells. These were associated with decreased MMP2 activity, as demonstrated by gel zymography. Rat astrocyte-conditioned media increased expression of MMP2 in ENU15645 cells and increased in vitro cell invasion of ENU1564 and ENU1564-TIMP2 cells. Blockage of ERK1/2 phosphorylation by treatment with PD98059 decreased the expression of MMP2 in cancer cells grown in rat astrocyte-conditioned media. We determine that MMP2 plays a role in in vivo development of breast cancer brain metastases. Additionally, we conclude that astrocytes are associated with expression of MMP2 in cancer cells via ERK1/2 signaling pathway.

Chromobacterium violaceum (CV) produces a violet color pigment known as Violacein. It has been reported that violacein has anticancer activity. This compound is produced by CV a gram-negative facultatively anaerobic bacterium found in soil and water environmental samples. The purpose of this study was to determine the effect of purified violacein on select cancer cell lines. Violacein used in this study was purified from CV strain (14N23), a strain isolated from environmental samples collected in the Tennessee Copper Basin. The previous reports used a crude extract preparation of violacein; thus, it was of interest to determine the effect of the pure compound on cancer cell growth was similar to that of the crude extracts. The compound purified following the method of Mehta, et al. was exposed to cancer cells and cell death assessed using the Alamar Blue procedure. It was found that violacein had no effect on A549, BT549, and PC3 cancer cell growth; however, there was a significant effect on Colo-320 cancer cells. It was concluded that further studies are required to assess the effect of violacein on enzymes and proteins involved in the cancer cell apoptotic pathways. Such studies will explain why cancer cell death was observed in certain cancer cells and not others.

Cancer is the second leading cause of death worldwide. Breast and lung cancers are highly diffused in population with a negative impact on mortality due to the high rate of dissemination and chemoresistance. The understanding of new molecular mechanisms and the introduction of new drugs are key elements in the oncology research. In cancer cells, a deregulation of metabolism and redox balance has been reported, with an increase of the detoxifying systems. High levels of reactive species may induce tumor cells to die. Many chemotherapeutic agents act in this way, shifting the balance toward an overwhelming of the endogenous scavenger systems. Nitric oxide (NO) is a Reactive Oxygen Species (ROS) member having proneoplastic and antineoplastic activity, depending on its concentration and cellular context. The first aim of this thesis was to investigate the antitumor activity of NO-donor (termed metal-nonoate) on A549 cells, a cell line representative of lung cancer. In this study, we observed cytotoxic activity of metal-nonoate and a reduction of the main hallmarks of tumor malignancy such as tumor cell clonogenicity, invasion and tumor angiogenesis. Upon NO donor treatment in A549, apoptosis pathway was activated with an upregulation of p53 and accumulation of cytochrome c. The mechanisms underlying these events were the phosphorylation of ERK1/2 and a burst of ROS production. The second part of my thesis addressed the role in tumor angiogenesis of aldehyde dehydrogenase (ALDH), a detoxifying enzyme that catalyzes the oxidation of aldehydes, a class of metabolic intermediates of tumor cell metabolism. The isoform ALDH1A1 is an important marker of breast cancer stemness. In this second study we observed a link between stemness status of breast cancer cells (MCF-7) and tumor angiogenesis, mediated by ALDH1A1. In MCF-7 overexpressing ALDH1A1, we observed a greater expression of angiogenic factors, such as HIF-1α and VEGF compared to wild type cells. In a panel of in vitro co-culture experiments, we demonstrated a promotion of endothelial cells recruitment that was dependent on VEGF expression, which was released by MCF-7 over-expressing ALDH1A1. This result was associated to in vivo tumor growth and angiogenesis in MCF-7 xenograft models, evaluated by blood flow and microvessel density (MVD). The intratumoral level of HIF-1α and VEGF was higher in MCF-7 tumor enriched of ALDH1A1, in which we also observed a greater expression of stemness markers, demonstrating a correlation between tumor stemness and tumor angiogenesis. In conclusion, our findings indicate that deregulation of tumor cell metabolism, a hallmark of cancer, is functionally linked with tumor angiogenesis and its modulation appears a promising strategy for cancer treatment.

In most randomised controlled trials (RCTs), a large number of patients need to be followed over many years, for the clinical benefit of the drug to be accurately quantified (1). Using an early proxy, or a surrogate endpoint, in place of the direct endpoint of overall survival (OS) could theoretically shorten the duration of RCTs and minimise the exposure of patients to ineffective or toxic treatments (2, 3). This thesis examined the relationship between surrogate endpoints and OS in metastatic colorectal cancer (CRC), advanced non-small cell lung cancer (NSCLC) and metastatic breast cancer (MBC). A review of the literature identified 144 RCTs in metastatic CRC, 189 in advanced NSCLC and 133 in MBC. The publications were generally of poor quality with incomplete reporting on many key variables, making comparisons between studies difficult. The introduction of the CONSORT statement was associated with improvements in the quality of reporting. For CRC (337 arms), NSCLC (429 arms) and MBC (290 arms) there were strong relationships between OS and progression free survival (PFS), time to progression (TTP), disease control rate (DCR), response rate (RR) and partial response (PR). Correlation was also demonstrated between OS and complete response (CR) in CRC and duration of response (DOR) in MBC. However, while strong relationships were found, the proportion of variance explained by the models was small. Prediction bands constructed to determine the surrogate threshold effect size indicated that large improvements in the surrogate endpoints were needed to predict overall survival gains. PFS and TTP showed the most promise as surrogates. The gain in PFS and TTP required to predict a significant gain in overall survival was between 1.2 and 7.0 months and 1.8 and 7.7 months respectively, depending on trial size and tumour type. DCR was a better potential predictor of OS than RR. The results of this study could be used to design future clinical trials with particular reference to the selection of surrogate endpoint and trial size.

Bronchoalveolar lavage (BAL) is an established method providing diagnostic support and evaluation of disease activity in interstitial lung disease (ILD). The aims of the present investigation were 1) to study the inflammatory response in pneumonitis evoked by irradiation. 2) to evaluate how well lung tissue inflammation is reflected in BAL findings. 3) to study the effect of smoking on radiation-induced pneumonitis. BAL was performed in 21 patients (11 smokers, 10 non-smokers) who were treated for breast cancer, stage 1 (TjMaNq) by post-surgery irradiation to an accumulated target dose of 56 Gy. It was founa that irradiation induced an alveolitis in the non-smoking patient group while the smoking patients did not differ from their smoking controls. The alveolitis in non-smokers was characterized by an increase in lymphocytes, mast cells and elevated concentrations of hyaluronan (HA), and fibronectin (FN). Three of the non-smoking patients had chest X-ray infiltrates indicating the presence of pneumonitis. An animal experimental model for radiation-induced pneumonitis and fibrosis was established in rats, allowing comparative analysis of BAL fluid and morphology. In the rat model a divergence was noted between the differential cell counts in BAL and cells observed in the interstitial tissue, which was most notable for neutrophils (PMN) and mast cells whereas there was a good correlation between HA content in BAL and HA deposition in the lung tissue. A marked infiltration of intraseptally-located mast cells occurred during the pneumonitis-phase, and this increase was paralleled by a deposition of HA in the interstitial tissue. Histochemical fixation and staining properties of the mast cells revealed that the majority of these cells were of connective tissue mast cell type (CTMC). Compound 48/80, a mast cell secretagogue, significantly altered the HA content both in BAL and in lung tissue in the irradiated animals. Regular treatment throughout the whole experimental period induced depletion of mast cell granules and a decrease in HA deposition whereas 48/80 treatment during the pneumonitis phase enhanced HA deposition. A rat model with smoke exposure was developed, and the effect of cigarette smoke on radiation-induced inflammation was studied. Rats that smoked 3 weeks prior to irradiation and continued to smoke throughout the observation period (7 weeks) had a significantly reduced inflammatory response compared to irradiated non-smoking rats. The most prominent BAL findings in the smoke-exposed rats were a decrease in PMN, mast cells and a decrease in HA. In conclusion, irradiation induces an alveolitis characterized mainly by mononuclear cells. Mast cells seem to be of importance in the remodelling of the connective tissue in the radiation-induced inflammatory response. Hyaluronan is an important component in the early connective tissue response preceding later collagen deposition, and its interstitial deposition is very well reflected in BAL. Moreover, tobacco-smoke suppresses the radiation-induced inflammation with a decreased recruitment of effector cells including mast cells.

Diss. (sammanfattning) Umeå : Umeå universitet, 1992, härtill 5 uppsatser.

digitalisering@umu

Tumour microenvironment is a complex network of cells, including immune cells, with a role in tumorigenesis and metastatic spread. Among the immune cells, M2-polarized Tumour-Associated-Macrophages (M2-TAMs) show immunosuppressive activities by expressing inflammatory molecules, thus promoting tumorigenesis. Triple-Negative Breast Cancer (TNBC), lacking both hormone receptors (i.e. Progesterone Receptor [PgR] and Estrogen Receptor [ER]) and Human Epidermal growth factor Receptor 2 (HER2), is associated with poor prognosis and high probability of distant metastases. In TNBC, a large number of infiltrating M2-TAMs is positively correlated to higher risk of metastases and lower rates of Disease-Free Survival” (EFS) and Overall Survival (OS). Prune-1 belongs to DHH (Asp-His-His) phosphoesterase superfamily with an exopolyphosphatase activity. The overexpression of Prune-1 is correlated with metastases and poor prognosis in several tumours including Breast Cancer (BC). Prune-1 was also found to induce Epithelial-Mesenchimal-Transition (EMT) and metastatic dissemination throught the enhancement of canonical TGF-β signaling by counterbalancing its inhibition operated by NDPK-A. Further evidences suggested that lung cancer progression is driven by Prune-1 through the canonical WNT signaling pathway in both autocrine and paracrine manner via Wnt3a cytokine secretion, thus suggesting a potential role for Prune-1 also in extracellular environment. Here, we identified Prune-1 protein as overexpressed in TNBC patients and positively correlated to lynphnode metastases, inflammatory pathways and infiltrating M2-TAMs (CD68+/CD163+). Furthermore, we developed a Genetically Engineered Mouse Model (GEMM) of metastatic TNBC over-expressing both Prune-1 and Wnt-1 in mammary glands (MMTV-Prune1/Wnt1). We found Prune-1 to recruit TAMs and to enhance their polarization toward a pro-tumorigenic M2-phenotype in the tumour microenvironment, thus promoting lung metastases in this GEMM. We also show that Prune-1 takes part to the communication between TNBC cells and TAMs through intracellular pathways activation (i.e., TGF-b and NF-kB), but also in a paracrine manner by inducing the secretion of inflammatory cytokines (e.g., IL-17F and IL-28) and modulating the exosome protein contents. Finally, we found an anti-Prune-1 drug (AA7.1) with the ability to reduce the primary tumour growth by reducing the percentage of M2-TAMs in orthotopic xenograft immunocompetent models of TNBC. In conclusion, our GEMM of metastatic TNBC driven by Prune-1 will be a useful source for future immunotherapy pre-clinical trials targeting M2-polarized TAMs within the tumour microenvironment to inhibit distant metastases in TNBC with poor prognosis.

DNA-damaging chemotherapeutics are a consistently employed for the treatment of cancer, and are regularly part of first-line combination therapeutic regimens. However, these regimens often display limited activity in cancers including of the lung and breast. Novel therapeutic strategies are urgently required. Understanding the molecular mechanisms regulating DNA-damaging chemotherapeutics activity will lead to such novel strategies. Activating transcription factor 3 (ATF3) is a stress inducible gene that plays a significant role in regulating cellular stress including DNA damage. This thesis investigates the role of ATF3 in mediating the cytotoxic effects of two commonly employed DNA-damaging chemotherapeutics, cisplatin and doxorubicin. This study also identifies other independent ATF3 inducing agents in potential novel combination therapeutic strategies. In this study, cell line models of cisplatin-resistance were generated independently from two non-small cell lung cancer (NSCLC) cell lines, Calu6 and H23. Full transcriptome RNA-sequencing analysis identified ATF3 as the most highly dysregulated apoptosis regulatory gene in the cisplatin-resistant compared to their respective parental cell lines following treatment with cisplatin. Further characterization identified cisplatin induced activation of JNK as the key regulator of ATF3 induction in these cell lines. Restoring JNK activity resulted in induced ATF3 expression and re-sensitization of the resistant cell lines to cisplatin treatment. FDA-approved 1200 compound library screens were employed to identify agents that can enhance cisplatin cytotoxicity as well as whose cytotoxicity was dependent on ATF3 expression. Vorinostat, an HDAC inhibitor, was identified in both screens and importantly displayed synergistic cytotoxicity in combination with cisplatin. In addition, ATF3 was also induced with treatments of the DNA damaging agent doxorubicin in these NSCLC cell lines including in the cisplatin resistant models. This work suggested a different mechanism of ATF3 induction by doxorubicin and the potential role of ATF3 in mediating doxorubicin cytotoxicity was further investigated in breast cancer cells. Doxorubicin robustly increased ATF3 expression in human breast cancer cell lines and tumor tissue ex-vivo. Loss of ATF3 in MEFs resulted in reduced sensitivity to doxorubicin treatment compared to wild-type MEFs. Employing the same library screen as above, compounds with ATF3 dependent mechanisms that could enhance doxorubicin cytotoxicity were identified. Vorinostat, as well as, the nucleoside analogues trifluridine and 6-mercaptopurine induced ATF3 expression and enhanced doxorubicin cytotoxicity. This study demonstrated that ATF3 plays an important role in mediating the cytotoxic effect of the DNA-damaging chemotherapeutics cisplatin and doxorubicin. It also provides rationale for ATF3 as a potential therapeutic target, and for the incorporation of ATF3 inducing agents in novel combination therapeutic strategies.

Al momento sono stati classificati più di 100 forme di cancro secondo il tipo di cellula inizialmente affetta. Le patologie tumorali sono caratterizzate da una crescita cellulare incontrollata che porta alla formazione di grumi o masse di tessuto che danneggiano l organismo. Lo sviluppo di nuovi farmaci antitumorali è una delle attività prioritarie nell'ambito della ricerca chimico-farmaceutica, non solo per l alta incidenza ma anche per i numerosi inconvenienti legati ai farmaci in uso. Il progetto di ricerca è stato indirizzato al disegno, sintesi e valutazione biologica di nuovi farmaci antitumorali derivati da una serie di composti dotati di notevole attività apoptotica e capaci di bloccare il ciclo cellulare. Questi derivati presentano strutture 1,4-diossepaniche, strutture di tipo para-nitrobenzensolfonammidico e strutture fenilpirrolidiniche. Le molecole sintetizzate sono state testate contro le linee cellulari di cancro di polmone (AC49), seno (MCF-7) e prostata (PC-3) mostrando la loro migliore attività nei confronti di quest ultima. I composti più attivi hanno mostrato valori di IC50 compresi tra 20,27 e 31 microM.

The major advantage of position sensitive radiation detector systems lies in their ability to non invasively map the regional distribution of the emitted radiation in real-time. Three of such detector systems were studied in this thesis, gamma-cameras, positron cameras and CMOS image sensors. A number of physical factors associated to these detectors degrade the qualitative and quantitative properties of the obtained images. These blurring factors could be divided into two groups. The first group consists of the general degrading factors inherent to the physical interaction processes of radiation with matter, such as scatter and attenuation processes which are common to all three detectors The second group consists of specific factors inherent to the particular radiation detection properties of the used detector which have to be separately studied for each detector system. Therefore, the aim of this thesis was devoted to the development of computational methods to enable quantitative molecular imaging in PET, SPET and in vivo patient dosimetry with CMOS image sensors.

The first task was to develop a novel quantitative dual isotope method for simultaneous assessments of regional lung ventilation and perfusion using a SPET technique. This method included correction routines for photon scattering, non uniform attenuation at two different photon energies (140 and 392 keV) and organ outline. This quantitative method was validated both with phantom experiments and physiological studies on healthy subjects.

The second task was to develop and clinically apply a quantitative method for tumour to background activity uptake measurements using planar mammo-scintigraphy, with partial volume compensation.

The third stage was to produce several computational models to assess the spatial resolution limitations in PET from the positron range, the annihilation photon non-collineairy and the photon depth of interaction.

Finally, a quantitative image processing method for a CMOS image sensor for applications in ion beam therapy dosimetry was developed.

From the obtained phantom and physiological results it was concluded that the methodologies developed for the simultaneous measurement of the lung ventilation and perfusion and for the quantification of the tumour malignancy grade in breast carcinoma were both accurate. Further, the obtained models for the influence that the positron range in various human tissues, and the photon emission non-collinearity and depth of interaction have on PET image spatial resolution, could be used both to optimise future PET camera designs and spatial resolution recovery algorithms. Finally, it was shown that the proton fluence rate in a proton therapy beam could be monitored and visualised by using a simple and inexpensive CMOS image sensor.

Hintergrund: Das Mammakarzinom stellt weltweit die häufigste maligne Tumorerkrankung der Frau dar und wird immer noch größtenteils als primär systemische Krebsform angesehen. Nach Primärbehandlung werden 5-Jahresüberlebensraten von 80% erreicht. Jedoch überleben Patientinnen, bei denen ein Stadium IV vorliegt, im median nur 20-30 Monate. 5-15% dieser Patientinnen weisen dabei einen isolierten metastatischen Befall der Lunge auf, der als Oligometastasierung im Sinne eines stabilen Zwischenstadiums der Erkrankung angesehen werden kann und somit einer lokalen Therapie zugänglich ist. Etliche Studien weisen darauf hin, dass gerade diese Frauen von einer Resektion ihrer Lungenmetastasen deutlich mehr profitieren können, als von medikamentöser systemischer Therapie allein. Zudem kann das Rezeptorverhalten (Östrogen-, Progesteron-, HER2-Rezeptor) zwischen primärem Mammakarzinom und dessen Metastasen differieren, was in bisher noch nicht geklärtem Umfang Änderungen des Behandlungsschemas zur Folge hat. Frage- und Zielstellung: Ziel der vorliegenden Arbeit ist es, das Outcome von Patientinnen mit pulmonal metastasiertem Mammakarzinom, deren Lungenmetastasen ausschließlich mit einer neuen parenchymsparenden 1318-nm-Lasertechnik reseziert wurden, im Verlauf zu untersuchen und unabhängige prognostische Faktoren zu identifizieren. Weiterhin soll der Nachweis einer Rezeptordiskordanz speziell für pulmonale Fernmetastasen erbracht und aufgezeigt werden, in welchen Größenordnungen mit solchen Rezeptorwechseln zu rechnen ist. Patientinnen und Methoden: Im Rahmen dieser retrospektiven Studie wurde vom 01.01.1996 bis 31.12.2012 bei insgesamt 102 Patientinnen im Alter von 33 bis 78 Jahren und einem Durchschnittsalter von 58 Jahren eine kurative pulmonale Laser-Metastasenresektion mit systematischer Lymphadenektomie vorgenommen. Vorgegebene Einschlusskriterien waren die vollständige Resektion und Kontrolle des Primärtumors sowie das Fehlen von extrapulmonalen/-thorakalen Metastasen bzw. deren präoperative erfolgreiche Therapie. Eine Limitierung bezüglich der Zahl der Lungenmetastasen wurde nicht vorgegeben, allerdings mussten die technische Resektabilität und die funktionelle Operabilität aus der präoperativen Diagnostik ableitbar sein. Mit Hilfe der Kaplan-Meier-Methode wurde das Gesamtüberleben des Patientenkollektivs sowie ausgewählter Subgruppen analysiert. Das Cox-Proportional-Hazard-Modell wurde verwendet, um im uni- und multivariaten Verfahren prognostische Faktoren zu ermitteln. Zum Vergleich des Rezeptorstatus von Primärtumor und Metastasen kam der McNemare-Test zum Einsatz. Eine statistische Signifikanz wurde bei p-Werten von < 0,05 angenommen. Ergebnisse: Insgesamt wurden 936 Lungentumore entfernt, von denen sich nach histopathologischer Sicherung 716 als Metastasen des anamnestisch bekannten Mammakarzinoms erwiesen. Die Anzahl reichte von einer solitären Metastase bis zu 61 zweizeitig entfernten Metastasen (durchschnittlich 7 pro Patientin). Die Lobektomierate betrug 0,98% (n=1). In 7,8% (n=8) der Fälle waren zusätzlich lappensparende Laser-Segmentresektionen möglich. R0-Resektionen konnten bei 73,5% (n=75) der Patientinnen erreicht werden. Das mediane Gesamtüberleben betrug 43 Monate, die 5-Jahresüberlebensrate belief sich auf 46,1%. Als unabhängige prognostische Faktoren konnten der Resektionsstatus (p=0,02), der intrathorakale Lymphknotenbefall (p=0,001) und die Expression des Östrogenrezeptors (p=0,018) nachgewiesen werden. Das Risiko zu versterben war bei tumorbefallenen Lymphknoten und bei fehlender Ausprägung des Östrogenrezeptors 3,2- bzw. 2-fach erhöht. Die Anzahl der resezierten Metastasen, die Art des Lungenbefalls (uni-/bilateral), das krankheitsfreie Überleben nach Primär-Operation ( 36 Monate) und die Expression des Progesteronrezeptors hatten keinen signifikanten Einfluss auf das Überleben. Angaben zum primären und metastatischen Hormonrezeptor- bzw. HER2-Status waren bei 88,2% (n=90) bzw. 62,7% (n=64) der Patientinnen verfügbar. Es fanden sich Diskordanzraten bzgl. des Östrogen-, Progesteron- und HER2-Rezeptors von 26,7%, 41,1% bzw. 28,1%. Eine Signifikanz der Abweichung zwischen Primärtumor und Metastasen konnte lediglich für den Östrogenrezeptor nachgewiesen werden (p=0,002). In einer Nebenbetrachtung der vorliegenden Arbeit konnten bei 157 Mammakarzinom-Patientinnen mit neu aufgetretenen, radiologisch detektierten Lungenrundherden in 65,6% der Fälle Metastasen des Mammakarzinoms histologisch gesichert werden. Bei den übrigen Befunden handelte es sich um andere therapiebedürftige maligne Tumore und zu etwa 20% um benigne Befunde. Schlussfolgerungen: Die vorliegenden Ergebnisse bekräftigen den positiven Einfluss der Lungenmetastasektomie auf das Überleben ausgewählter Mammakarzinom-Patientinnen mit isolierter pulmonaler Oligometastasierung. Dabei können mit der Anwendung der parenchymsparenden 1318 nm -Lasertechnik, auch bei Vorhandensein von multiplen und beidseitigen Lungenmetastasen, in größerem Umfang als bisher berichtet, vollständige Resektionen ohne wesentlichen Funktionsverlust und somit guter Lebensqualität erreicht werden. Ungeachtet höherer Zahlen resezierter pulmonaler Metastasen werden gegenüber konventionellen Operationstechniken, auch beim pulmonal metastasierten Mammakarzinom, vergleichbare Überlebensraten erreicht. Die Anzahl der präoperativ diagnostizierten Lungenmetastasen sollte daher einen geringen Einfluss auf die Indikationsstellung zur Operation haben, weshalb diesbezüglich eine Erweiterung der Einschlusskriterien sinnvoll erscheint. Eine R0-Resektion konnte erneut als wichtigster prognostischer Parameter bestätigt werden und sollte deshalb stets oberstes Ziel des Operateurs sein. Das wesentlich schlechtere Outcome unvollständig operierter Patientinnen sowie der Vergleich mit der Literatur zur alleinigen systemischen Therapie zeigen, dass das analysierte Patientenkollektiv von einer Resektion der pulmonalen Mammakarzinom-Metastasen deutlich mehr profitieren kann, als von medikamentöser Behandlung allein. Ein intrathorakaler Lymphknotenbefall wurde, nach unserem Wissen, erstmals bei Patientinnen mit isolierten Lungenmetastasen eines Mammakarzinoms, trotz radikaler Ausräumung, als signifikante negative Einflussgröße auf das Überleben nachgewiesen. In Anlehnung an die Therapie des Lungenkarzinoms sollte trotz dessen, zumindest bis zum Vorliegen weiterführender Studien, standardmäßig eine intraoperative systematische Lymphadenektomie durchgeführt werden. Bei positivem Tumornachweis ist eine komplette Lymphknotendissektion zu erwägen, um keine Patientin von einer potenziell kurativen Therapie auszuschließen. Den vorliegenden Ergebnissen zufolge, darf des Weiteren speziell bei Verdacht auf pulmonale Mammakarzinom-Metastasen nicht von einer Konstanz der Expression der Steroidhormon- bzw. HER2-Rezeptoren, insbesondere der des Östrogenrezeptors, ausgegangen werden. Änderungen zum primären Befund treten dabei in relevanten Größenordnungen auf. Die Bestimmung eines aktuellen Rezeptorstatus sollte nach Metastasektomie obligat durchgeführt werden. Bezüglich der Frage des Ursprungs pulmonaler Rundherde bei bekanntem Mammakarzinom kann darüber hinaus durch deren Resektion mit nachfolgender histopathologischer Analyse sicher zwischen Metastasen, Lungenkarzinomen und benignen Tumoren differenziert werden. Insgesamt ermöglicht dies konkrete Therapieentscheidungen zu treffen. Um Patientinnen jedoch in zeitlich limitierter oligometastatischer Tumorausbreitung zu diagnostizieren und einer bestmöglichen Therapie, einschließlich der Resektion, zuzuführen, ist zufolge unserer Daten sowie der neueren Literatur eine konsequente, engmaschige und zudem apparative Nachsorge notwendig. Dieser Problematik wird gegenwärtig in den aktuellen Leitlinien nicht adäquat Rechnung getragen, da sich die Autoren auf ältere, heutzutage kritisch zu hinterfragende Analysen beziehen. Als Limitationen der vorgelegten Arbeit sind das retrospektive Studiendesign und die Form der Kohortenanalyse, die uneinheitliche Bestimmung des primären Rezeptorstatus sowie die Heterogenität der postoperativen Anschlusstherapien anzusehen. Zukünftig sind größere, multizentrische und randomisierte Studien notwendig, um weiterführende Daten zu generieren und die pulmonale Lasermetastasektomie beim Mammakarzinom im Rahmen multimodaler Therapien möglicherweise weiter zu etablieren sowie den Wert einer erweiterten Nachsorge zu evaluieren
Background: Breast carcinoma is the most common type of cancer in women worldwide and is still regarded as a systemic disease. After primary treatment five-year survival rates around 80% are reported. However, the mean survival time of stadium-IV classified patients is 20-30 months. 5-15% of patients appear with isolated metastases of the lungs which can be considered as an oligometastatic and, therefore, stable intermediate stage in disease process. Several studies point out that especially these women are more likely to benefit from resection of lung metastases than from systemic therapy alone. Furthermore, there is the possibility of a discordant expression of typical receptors (Estrogen-, Progesterone- and HER2-receptor) between primary breast cancer and its paired metastases. As a result a change in treatment regimen might be necessary. Objective: The aim of the present study was to evaluate long-time survival of patients with lung metastases from breast cancer who have been operated exclusively with a new parenchyma-saving and lobe-sparing 1318-nm-lasertechnique. Additionally, the identification of independent prognostic factors was of interest. Furthermore, existence and magnitude of receptor discordance, specifically for distant pulmonary metastases, should be proved. Patients and methods: Within this retrospective study between 1996 and 2012 102 patients (mean age 58; range 33-78 years) underwent curative laser metastasectomy and systematic lymphadenectomy. Inclusion criteria were complete resection of primary breast cancer and absence of extrapulmonary/-thoracal metastases or its previous total treatment. Although there were no limitations regarding the number of metastases, technical resectability and functional operability had to be assumed after the preoperative diagnostics. Kaplan-Meier-analysis was performed to assess overall survival in all patients and selected subgroups. Uni- and multivariate analyses of prognostic factors were performed using the Cox-proportional-hazard model. Comparison of the receptor status of primary breast cancer and paired lung metastases was assessed by the McNemare method. Significant results were assumed if p-values were <0.05. Results: In total 936 intrapulmonary nodules had been resected, including 716 histopathologically confirmed breast cancer metastases. The amount reached from a single metastasis up to 61 two-staged removed pulmonary nodules (mean 7 per patient). The lobectomy rate was 0.98%. In 7.8% of all cases segment-resections, also performed by laser, were possible. Complete resection was achieved in 73.5% (n=75). The median overall survival time was 43 months and the five-year survival rate was 46.1%. As independent prognostic factors resection status (p=0.02), involvement of intrathoracal lymph nodes (p=0.001) and expression of estrogen receptor (p=0.018) were identified. The mortality rate in case of lymph node involvement and negative estrogen receptor status was increased by 3.2- and 2-fold, respectively. The number of resected metastases, type of lung affection (uni-/bilateral), disease free interval after primary breast surgery ( 36 months), and expression of progesterone receptor had no significant influence on survival. Data concerning the primary and metastatic hormone receptor- and HER2-status were available in 88.2% (n=90) and 62.7% (n=64) of all cases, respectively. Discordant results appeared in 26.7%, 41.1%, and 28.1% regarding the estrogen-, progesterone- and HER2-receptor. Significance of these findings had only been proved for estrogen receptor (p=0.002). A subanalysis of the present study revealed that 65.5% of 157 breast cancer patients who presented with newly occurred radiologically detectable pulmonary nodules had histopathologically confirmed paired metastases. The remaining results showed malignancies other than known breast cancer, and in approximately 20% of all cases there were benign lesions. Conclusions: The results of the present study emphasize the favorable effect of the lung metastasectomy on survival of selected breast cancer patients with isolated pulmonary oligometastatic disease. Via the use of the parenchyma-saving 1318nm-lasertechnique even in case of distinct and bilateral pulmonary metastatic involvement, increased rates of complete resection without substantial loss of lung function can be achieved. Therefore, an adequate quality of life is provided. In comparison with conventional surgery practices, this procedure creates similar survival rates despite higher numbers of resected lung metastases. That is why the number of preoperatively diagnosed metastases should have little influence on decision upon surgery. Thus, an extension of inclusion criteria seems reasonable. Again, complete resection appeared as one of the most important prognostic parameters and should, therefore, be the main objective of the surgeon. The poorer outcome for women with incomplete resections and the results of studies on systemic therapy implicate once more that breast cancer patients are more likely to benefit from the resection of their pulmonary metastases than from medical treatment alone. Furthermore, for the first time according to our knowledge, despite radical excision intrathoracal lymph node involvement has been proved as a significant negative predictive determinant in a collective of patients with isolated pulmonary metastases of breast cancer. Nevertheless, an intraoperative systematic lymph node sampling should be considered, at least until further studies are presented. In reference to the surgical approach of lung carcinoma, as circumstances require, a complete lymph node dissection should be performed to provide potential curative treatment to those affected. Moreover, according to the present findings, in case of the appearance of lung metastases the constancy of metastatic steroid hormone- and HER2 receptor expression, especially of the estrogen receptor, cannot always be assumed. Changes in comparison to the primary carcinoma appear in a relevant number of cases. Thus, the current metastatic receptor status should be evaluated obligatorily after pulmonary metastasectomy. Regarding the origin of pulmonary nodules of patients with history of breast cancer, their surgical resection with subsequent histopathological analysis can reliably differentiate between metastases, lung carcinoma or benign tumors. Altogether this facilitates specific and accurate treatment decisions. However, to identify patients with a limited and stable oligometastatic state of disease and to introduce optimal treatment, including surgical resection, an early, continuous, and also instrument-based follow-up is necessary. This matter is still only slightly taken into account, while the authors of the current guidelines refer to out of date studies, which have to be seen critically. As limitations of the present investigation, the retrospective study design, inconsistent evaluation of the primary receptor status, and also heterogeneity of postoperative medical therapy must be mentioned. In the future larger, multicentric, prospective, randomized trials are necessary to acquire further data, to conceivably continue to establish the pulmonary laser metastasectomy in multimodal therapy settings and also to determine the value of an extended follow-up

This diploma thesis deals with various ways of treatment of selected oncological diseases and the effectiveness of treatment methods and evaluation of the influence of various factors influencing the survival of patients. The activity of individual healing processes is evaluated by survival analysis. The subjects of the study are patients with breast, lung and prostate cancer. The survival analysis considers the sex of the patient, the age and stage of his illness, and other factors to avoid distorted results. The aim of the work is to find out the effects of selected therapeutic procedures on patients' health and to identify factors that have a significant impact on the survival of patients. The data for the diploma thesis was provided by the Institute of Health Information and Statistics of the Czech Republic, the Statistical Office, the National Cancer Register (NOR), the US SEER database and the German Breast Cancer Study.

Medical imaging is a powerful tool for clinical practice allowing in-vivo insight into a patient’s disease state. Many modalities exist, allowing for the collection of diverse information about the underlying tissue structure and/or function. Traditionally, medical professionals use visual assessment of scans to search for disease, assess relevant disease predictors and propose clinical intervention steps. However, the imaging data contain potentially useful information beyond visual assessment by trained professional. To better use the full depth of information contained in the image sets, quantitative imaging characteristics (QICs), can be extracted using mathematical and statistical operations on regions or volumes of interests. The process of using QICs is a pipeline typically involving image acquisition, segmentation, feature extraction, set qualification and analysis of informatics. These descriptors can be integrated into classification methods focused on differentiating between disease states. Lung cancer, a leading cause of death worldwide, is a clear application for advanced in-vivo imaging based classification methods. We hypothesize that QICs extracted from spatially-linked and size-standardized regions of surrounding lung tissue can improve risk assessment quality over features extracted from only the lung tumor, or nodule, regions. We require a robust and flexible pipeline for the extraction and selection of disease QICs in computed tomography (CT). This includes creating an optimized method for feature extraction, reduction, selection, and predictive analysis which could be applied to a multitude of disease imaging problems. This thesis expanded a developmental pipeline for machine learning using a large multicenter controlled CT dataset of lung nodules to extract CT QICs from the nodule, surrounding parenchyma, and greater lung volume and explore CT feature interconnectivity. Furthermore, it created a validated pipeline that is more computationally and time efficient and with stability of performance. The modularity of the optimized pipeline facilitates broader application of the tool for applications beyond CT identified pulmonary nodules. We have developed a flexible and robust pipeline for the extraction and selection of Quantitative Imaging Characteristics for Risk Assessment from the Tumor and its Environment (QIC-RATE). The results presented in this thesis support our hypothesis, showing that classification of lung and breast tumors is improved through inclusion of peritumoral signal. Optimal performance in the lung application achieved with the QIC-RATE tool incorporating 75% of the nodule diameter equivalent in perinodular parenchyma with a development performance of 100% accuracy. The stability of performance was reflected in the maintained high accuracy (98%) in the independent validation dataset of 100 CT from a separate institution. In the breast QIC-RATE application, optimal performance was achieved using 25% of the tumor diameter in breast tissue with 90% accuracy in development, 82% in validation. We address the need for more complex assessments of medically imaged tumors through the QIC-RATE pipeline; a modular, scalable, transferrable pipeline for extracting, reducing and selecting, and training a classification tool based on QICs. Altogether, this research has resulted in a risk assessment methodology that is validated, stable, high performing, adaptable, and transparent.

Vitamin D3 must be metabolically activated by the liver to 25-hydroxyvitamin D3 (25OHD3) and then by the kidney 1alphahydroxylase (1alphaOHase) to become 1,25dihydroxyvitamin D 3 (1,25(OH)2D3). 1,25(OH)2 D3 is a potent inhibitor of tumor growth in vitro and in vivo. Recent studies indicate that metabolic activation of 1,25(OH) 2D3 also occurs in cancer cells such as breast cancer. Consequently, the major objective of this project was to determine if tumoral 25OHD 3-1alphahydroxylase modulates any or all of the stages of breast tumor progression without inducing the hypercalcemic side effects of 1,25(OH) 2D3. For this purpose we used the PyVMT breast cancer mouse model in which the oncoprotein, polyomamiddle T antigen (PyMT) is under the control of mouse mammary tumor virus LTR (MMTV LTR). Mice exhibited tumors restricted to the mammary epithelium progressing to the various stages of breast cancer. Animals were treated with either vehicle, 25OHD3 (2000 pM/24h) or 1,25(OH)2D3 (12pM/24h). Mice treated with the vitamin D precursor, 25OHD3, exhibited a marked reduction in tumor onset and growth comparable to the 1,25(OH)2D3 treated group. Furthermore, biomarkers of tumor progression were markedly reduced in 25OHD3 and 1,25(OH)2D3 animals as compared to vehicle-treated animals. However, mean circulating calcium concentrations remained unchanged in 25OHD3 treated animals but increased significantly in 1,25(OH)2D3 treated animals as compared to controls. Tumoral levels of 1,25(OH)2D3 in mice treated with 25OHD3 were increased 79% in comparison to vehicle control mice. Additionally, 25OHD3 and 1,25(OH)2D 3 treated animals had a significant decrease in the mean number of lung metastases per animal as compared to vehicle treated control animals. This study therefore suggests an important autocrine role of 1alphaOHase expression in breast tumor cells. Furthermore, accumulation of intra-tumoral 1,25(OH) 2D3 in response to 25OHD3 administration strongly suggests that locally produced 1,25(OH)2D3 plays a significant role in restraining tumor growth without inducing the hypercalcemic side effects associated with 1,25(OH)2D3.

Improvements in the medical field have given many cancer patients and survivors better odds of long-term survival. As more patients become survivors, the demand for psychological treatment becomes greater. The most prevalent concern of survivors is getting help with a psychosocial condition known as fear of recurrence (FOR). Prior to this study, few researchers had explored how having a more aggressive cancer influences the development of FOR. The purpose of this quantitative study was to determine whether cancer stage and type (a measurement of severity) are predictive of FOR development in the high-risk cancer groups lung and bronchus and female breast. The theoretical framework guiding this research was based on Mishel's theory of uncertainty in illness, which states that uncertainties about illness recurrence can cause survivors to experience breakdown in their lives (whether psychological and/or physical). The fear of cancer recurrence inventory (FCRI) survey was administered to 97 lung and bronchus and female breast cancer survivors; the survivors were asked to rate their level of discomfort about the possibility of a cancer recurrence. Data were analyzed using multiple linear regression. The results indicated that cancer type and severity both impacted the development and severity of FOR in lung and bronchus and female breast cancer survivors. Furthermore, regardless of the cancer type, stage of cancer, age of the survivor, or years in remission, survivors reported clinical levels of FOR in all areas of concern. Practitioners can use the current findings to work towards developing better intervention and treatment programs that promote quality survivorship and reduce the risk and rate of FOR in high risk cancer populations.

The aims of this thesis were to investigate the long-term effects of adjuvant tamoxifen treatment on breast cancer recurrence and mortality, cardiovascular disease, and the incidence of secondary cancer. Between 1982 and 1992, postmenopausal patients with early stage breast cancer were included in a randomized clinical study of 2 or 5 years of postoperative tamoxifen therapy. The trial was planned by the Swedish Breast Cancer Group, and it included 4610 patients. Follow-up on causes of death, hospitalizations and secondary cancers were obtained from national population-based registries.  All-cause mortality, breast cancer-specific mortality and mortality from coronary heart disease were decreased in the 5-year group, but the incidence of endometrial cancer was increased (Paper I). The incidence and mortality of cerebrovascular diseases were increased during the active treatment phase, and reduced after the active treatment (Paper II). Similar results were seen for subgroups of cerebrovascular diseases such as stroke and ischemic stroke. In the 5-year group, the morbidity from coronary heart disease was reduced during treatment but not after treatment was stopped (Paper III). This was the case also for heart failure and for atrial fibrillation/flutter. For secondary cancers the lung cancer risk was reduced, as well as the lung cancer mortality (Paper IV). An increased risk was observed for endometrial cancer, but appeared to decrease over time. The risk of contralateral breast cancer was reduced, with most of the reduction after treatment was stopped. For distance recurrences the risk was reduced both during treatment and a few years after treatment was stopped. The breast cancer mortality was also reduced, especially during the post-treatment phase.

BACKGROUND:Numerous microarray-based prognostic gene expression signatures of primary neoplasms have been published but often with little concurrence between studies, thus limiting their clinical utility. We describe a methodology using logistic regression, which circumvents limitations of conventional Kaplan Meier analysis. We applied this approach to a thrice-analyzed and published squamous cell carcinoma (SQCC) of the lung data set, with the objective of identifying gene expressions predictive of early death versus long survival in early-stage disease. A similar analysis was applied to a data set of triple negative breast carcinoma cases, which present similar clinical challenges.METHODS:Important to our approach is the selection of homogenous patient groups for comparison. In the lung study, we selected two groups (including only stages I and II), equal in size, of earliest deaths and longest survivors. Genes varying at least four-fold were tested by logistic regression for accuracy of prediction (area under a ROC plot). The gene list was refined by applying two sliding-window analyses and by validations using a leave-one-out approach and model building with validation subsets. In the breast study, a similar logistic regression analysis was used after selecting appropriate cases for comparison.RESULTS:A total of 8594 variable genes were tested for accuracy in predicting earliest deaths versus longest survivors in SQCC. After applying the two sliding window and the leave-one-out analyses, 24 prognostic genes were identified
most of them were B-cell related. When the same data set of stage I and II cases was analyzed using a conventional Kaplan Meier (KM) approach, we identified fewer immune-related genes among the most statistically significant hits
when stage III cases were included, most of the prognostic genes were missed. Interestingly, logistic regression analysis of the breast cancer data set identified many immune-related genes predictive of clinical outcome.CONCLUSIONS:Stratification of cases based on clinical data, careful selection of two groups for comparison, and the application of logistic regression analysis substantially improved predictive accuracy in comparison to conventional KM approaches. B cell-related genes dominated the list of prognostic genes in early stage SQCC of the lung and triple negative breast cancer.

SASH1 (SAM and SH3 domain-containing protein 1) is a putative tumour suppressor functioning in the control of apoptosis and cellular proliferation. In summary this project has further characterised the role of SASH1 in genome maintenance, including DNA repair and apoptosis, which has implications on its links with tumourigenesis. In addition we have also identified that targeting SASH1 may be a novel cancer treatment target and shown that Chloropyramine may act as an effective anti-cancer agent through a SASH1-dependent pathway.

This thesis is focused on extracting meaningful statistics for the characterization and classification of biological, medical, and financial data and contains four chapters. The first chapter contains theoretical background on scaling and wavelets, which supports the work in chapters two and three. In the second chapter, we outline a methodology for representing sequences of DNA nucleotides as numeric matrices in order to analytically investigate important structural characteristics of DNA. This methodology involves assigning unit vectors to nucleotides, placing the vectors into columns of a matrix, and accumulating across the rows of this matrix. Transcribing the DNA in this way allows us to compute the 2-D wavelet transformation and assess regularity characteristics of the sequence via the slope of the wavelet spectra. In addition to computing a global slope measure for a sequence, we can apply our methodology for overlapping sections of nucleotides to obtain an evolutionary slope. In the third chapter, we describe various ways wavelet-based scaling may be used for cancer diagnostics. There were nearly half of a million new cases of ovarian, breast, and lung cancer in the United States last year. Breast and lung cancer have highest prevalence, while ovarian cancer has the lowest survival rate of the three. Early detection is critical for all of these diseases, but substantial obstacles to early detection exist in each case. In this work, we use wavelet-based scaling on metabolic data and radiography images in order to produce meaningful features to be used in classifying cases and controls. Computer-aided detection (CAD) algorithms for detecting lung and breast cancer often focus on select features in an image and make a priori assumptions about the nature of a nodule or a mass. In contrast, our approach to analyzing breast and lung images captures information contained in the background tissue of images as well as information about specific features and makes no such a priori assumptions. In the fourth chapter, we investigate the value of social media data in building commercial default and activity credit models. We use random forest modeling, which has been shown in many instances to achieve better predictive accuracy than logistic regression in modeling credit data. This result is of interest, as some entities are beginning to build credit scores based on this type of publicly available online data alone. Our work has shown that the addition of social media data does not provide any improvement in model accuracy over the bureau only models. However, the social media data on its own does have some limited predictive power.

La Ténascine-C (TNC) est une molécule de la matrice extracellulaire qui est anormalement surexprimée dans entre autres, le cancer du sein. Le but principal de ma thèse était de mieux comprendre les différentes contributions de la TNC, durant l’établissement et progression des cancers mammaires. Mes résultats acquis avec le modèle transgénique NeuNT soutiennent fortement la notion que la TNC exerce des rôles pléiotropiques et cruciaux à la fois à des stades précoces et à des stades ultimes de la progression tumorale mammaire. D'autre part, j’ai développé de nouvelles lignées cellulaires cancéreuses mammaires syngéniques à partir des deux modèles transgéniques. En utilisant des modèles murins et des approches in vitro, mon travail de doctorat a permis de mettre en lumière différentes contributions importantes de la TNC au cours de la carcinogenèse mammaire. Les analyses encore en cours permettront rapidement de mieux cerner les mécanismes moléculaires et cellulaires mis en jeu en aval de la TNC et impliqués dans la progression tumorale mammaire
The microenvironment, which comprises the extracellular matrix (ECM), plays instrumental roles during tumor formation and progression. Tenascin-C (TNC) is a major ECM component highly expressed in breast cancer, correlating with poor prognosis, tamoxifen resistance and lung metastasis formation. TNC exerts pleiotropic effects by promoting tumor cell survival, proliferation and invasion as well as angiogenesis , inflammation and metastasis. The main goal of my thesis was to try to comprehensively understand the several contributions of TNC during breast cancer establishment and progression to metastatic disease. My analyses revealed that the absence of TNC does not affect breast tumorigenesis in the MMTV-PyMT breast cancer mouse model, confirming a previous study. More interestingly, using the MMTV-NeuNT model, we show that TNC promotes primary tumor initiation and lung metastatic colonization. In the lung, TNC increases of cancer cell survival in intravascular metastases and promotes their progression. Additionally, we established breast cancer cell lines from MMTV-PyMT and MMTV-NeuNT mouse models that grow in vitro and are tumorigenic when re-Implanted in syngeneic, fully immuno-Competent mice. In this work we had shown that TNC participates in tumor initiation and in lung metastasis colonization in an ErbB2-Driven transgenic breast cancer mouse model. The established cell lines are alternative tools useful for in vivo and in vitro studies in breast cancer research

Le troisième Plan Cancer lancé en 2013 désigne la précocité du diagnostic comme l'un des enjeux majeurs pour l'amélioration de la prise en charge des patients. Malgré l’essor des modalités et des performances de l’imagerie médicale, il reste des défis à relever pour l’aide au diagnostic et optimiser le recourt à la biopsie.L’imagerie photonique et spécialement la fluorescence résolue spectralement a déjà été éprouvée pour la caractérisation ex vivo des tumeurs mammaires et pulmonaires, sans agent de contraste ou traitement des échantillons. Notre objectif est de caractériser les capacités d'un dispositif médical innovant, développé au laboratoire, utilisant une aiguille fibrée de faible calibre pour l’analyse spectrale de la fluorescence endogène de ces lésions in situ. Nos premiers travaux dans le cadre d’études précliniques et cliniques ont montré des différences significatives de signatures spectrales entre tumeurs bénignes et malignes ex vivo et in vivo. Nos résultats ont également mis en évidence les limites d’utilisation du dispositif, en termes de spécificité, pour certains types de lésions.Une étude secondaire a été entreprise sur des tumeurs mammaires afin d'identifier les entités tissulaires majeures à l'origine des signatures spectrales obtenues avec notre dispositif fibré. L'imagerie spectrale en microscopie confocale et seconde harmonique (SHG), en multiphoton, ont été mises en œuvre afin d’établir une cartographie de biomarqueurs endogènes des tissus mammaires. Nous avons confronter ses résultats aux données obtenues avec le dispositif d'aiguille fibrée afin de pouvoir le positionner non seulement comme une aide au diagnostic mais aussi comme une méthode prometteuse pour l’histopathologie in situ
The third Cancer Plan, launched in 2013, identifies early diagnosis as one of the major challenges for improving patient care. Despite the growth in medical imaging modalities and performance, challenges remain in diagnosis aid and optimizing the use of biopsy.Photonic imaging and especially spectrally resolved fluorescence has already been tested for the ex vivo characterization of breast and lung tumors, without contrast agent or sample processing. Our goal is to characterize the capabilities of an innovative medical device, developed in the laboratory, using a low-caliber fibered needle for the spectral analysis of the endogenous fluorescence of these lesions in situ. Our early work in preclinical and clinical studies showed significant differences in spectral signatures between benign and malignant tumors ex vivo and in vivo. Our results also highlighted the limits the device, in terms of specificity, for certain types of lesions.Another study was conducted on mammary tumors in order to identify the major tissue entities at the origin of the spectral signatures obtained with our fibered device. Spectral imaging in confocal and second harmonic microscopy (SHG), in multiphoton, has been implemented in order to establish a mapping of endogenous biomarkers of mammary tissues. We compare its results with the data obtained with the fibered needle device in order to position it not only as an aid to diagnosis but also as a promising method for in situ histopathology

Metastatic breast cancer treatment has seen few advances in recent years, yet treatment resistance continues to rise, causing disease recurrence. A pilot study was performed to determine the efficacy of ex vivo expansion and reprogramming of tumor-reactive immune cells from experimental metastatic tumor-sensitized mice. Also, phenotypic changes in tumors due to metastasis or tumor microenvironment influences were characterized. Metastatic neu+ mouse mammary carcinoma (mMMC) and its distant relapsing neu-antigen-negative variant (mANV) were investigated in FVBN202 mice. Tumor-reactive central memory CD8+ T cells and activated NK/NKT cells were successfully reprogrammed and expanded during 6-day expansion from mMMC- and/or mANV-sensitized mice, resulting in tumor-specific cytotoxicity. mMMC exhibited a flexible neu-expression pattern and acquired stem-like, tumorigenic phenotype following metastasis while mANV remained stable except decreased tumorigenicity. Myeloid-derived suppressor cell (MDSC) levels were not increased. Adoptive cellular therapy (ACT) with reprogrammed tumor-reactive immune cells may prove effective prophylaxis against metastatic or recurrent breast cancer.

Even though anti‐cancer chemotherapy has been continuously improved during the last decades. problems with adverse effects and drug resistance still constitutes a considerable obstacle and sets a demand for new effective treatment options. Tissue homeostasis in multi‐cellular organisms is maintained through intrinsic cell death, apoptosis, which removes unwanted or damaged cells. Disrupted apoptosis is an important factor in tumorgenesis and drug resistance, therefore induction or restoration of apoptotic pathways is also important for the treatment of cancer. Several naturally occurring bacterial toxins have the ability to induce apoptosis and could thus be candidates to complement or improve the therapeutic effect of other anticancer drugs. The bacterial toxins, adenylate cyclase (AC) toxin from Bordetella pertussis, α‐toxin from Staphylococcus aureus and verotoxin‐1 (VT‐1) from Escherichia coli were investigated for their ability to induce apoptosis in different tumor cell lines. Toxin induction of cell death was investigated by cell viability assays, end‐stage apoptosis induction by DNA‐fregmentation (TUNEL) assay. Toxin receptor expression and signal transduction pathways to apoptosis were investigated by flow cytometry, caspase enzyme activity assays and western blot. Immunohistochemistry was used for identification of toxin receptor expression in tumor tissue samples. AC‐toxin was cytotoxic and induced apoptosis in cultured malignant plural mesothelioma (MPM) and small‐cell lung cancer (SCLC) cells. Low‐toxic concentrations of AC‐toxin enhanced cisplatin cytotoxicity and apoptosis in both cell lines. MPM‐cells with acquired cisplatin resistance were more sensitive to α‐toxin than the less resistant parental MPM cell line. A low‐toxic concentration of α‐toxin re‐sensitized resistant MPM cells to cisplatin cytotoxicity by apoptosis induced through the mitochondrial pathway without detectable activation of common up‐stream apoptosis signalling proteins. VT‐1 was highly cytotoxic and induced apoptosis in globotriosylceramide (Gb3) ‐expressing glioma, breast cancer and non‐small‐cell lung cancer (NSCLC) cells but was not cytotoxic to non‐Gb3‐expressing cells. PPMP, an inhibitor of glucosylceramide synthesis which makes exposed cells unable to synthesize Gb3 rendered Gb3‐expressing cells resistant to VT‐1. MPM cells with acquired‐cisplatin resistance expressed Gb3 in contrast to the absent of expression in the less resistant parental cell line. Gb3, could however be up‐regulated by cisplatin in Gb3‐negative MPM‐cells. Presence of a low‐toxic concentration of VT‐1 potentiated cisplatin‐induced cytotoxicity and apoptosis in the cisplatin‐resistance MPM cell line. VT‐1 was a potent inducer of apoptosis, probably via stress‐induced Mitogen‐activated protein kinase (MAPK)‐signaling involving c‐Jun N‐terminal kinase (JNK) and p38, leading to disruption of the mitochondrial membrane integrety, activation of caspase‐9 and ‐3, and ultimately DNA fragmentation and cell death. Gb3 expression was demonstrated in clinical specimens of glioblastoma and breast cancer making these tumor types interesting for further VT‐1 studies. We conclude that bacterial toxins may be used to induce apoptosis in several types of cancer cells. Low concentrations of verotoxin‐1 and α‐toxin may potentially be used to overcome acquired cisplatin‐resistance in cancer patients.