published_or_final_version / abstract / Molecular Biology / Master / Master of Philosophy

published_or_final_version / abstract / Anatomy / Doctoral / Doctor of Philosophy

published_or_final_version / abstract / Pathology / Doctoral / Doctor of Philosophy

The immunoproteasome, known to play an important role in MHC class I antigen processing and presentation, have been linked to neurodegenerative diseases and hematological cancers. However, the pathophysiological functions of the immunoproteasome in these diseases are still not very well established. This can be attributed mainly to the lack of appropriate molecular probes that selectively target the immunoproteasome catalytic subunits. Herein, we report the development of a small molecular inhibitor (AM) that selectively targets the major catalytic subunit, LMP2, of the immunoproteasome. We show that the compound covalently modifies the LMP2 subunit with high specificity in human prostate cancer cell. AM was also shown to selectively inhibit the chymotrypsin-like activity of LMP2 subunit. More importantly, the anti-proliferative activity of AM is more pronounced in prostate cancer cells that highly express LMP2 without inducing toxicity in normal cells. These results implicate an important role of LMP2 in regulating cell growth of malignant tumors that highly express LMP2. Subsequently, the modes of action of AM were investigated. Prostate cancer cells that highly express LMP were shown to induce G2/M cell cycle arrest and apoptosis via PARP cleavage when treated with the compound. Similar to epoxomicin, the treatment of AM induced the accumulation of poly-ubiquitination in prostate cancer cells, which indicates the inhibition of proteolysis. However, unlike epoxomicin, the treatment of AM did not appear to inhibit the activation of inflammation. In conclusion, these results suggest that the LMP2 inhibitor, AM, may induce cytotoxicity prostate cancer cells that highly express LMP2 catalytic subunit in similar modes of action as epoxomicin but it does not involve the inflammatory pathway.

Prostate cancer (PCa) has been the leading cause of cancer death in men since1930. While studies pertaining to PCa have primarily focused on the disease and the subsequent side effects of treatment, psychological distress in this group has yet to be adequately addressed. The purpose of this dissertation was to: 1) conceptualize health related quality of life (HRQL) and health seeking behavior of men by describing lifestyle, cultural and health risks associated with being male, 2) evaluate the psychometric properties of the SF-12 Health Survey (SF-12) combined with the urinary and sexual portions of the UCLA PCa Index (UCLA-PCI), 3) investigate the psychological impact, coping styles and informational needs of a group of men diagnosed with PCa who have not yet undergone treatment. Men have been observed to underutilize health care services despite the fact that they are in poorer health, have higher mortality rates and lower life expectancies than women. Restricting emotions, being oriented toward success, having limited social networks and taking health risks are often associated with being male. Because incontinence and sexual dysfunction, the two most common side effects of PCa treatments impact men's quality of life, portions of the UCLA-PCI and the SF12 were analyzed. The psychometric analysis of the SF-12/UCLA-PCI and its three subcomponents confirmed the validity of the instrument. The SF-12 component had a Cronbach's alpha of 0.87, while the urinary and sexual subscales had a Cronbach's alpha of .86 and .91 respectively. All three scales were found to have good internal consistency. KEYWORDS: prostate cancer, health related quality of life, coping styles, psychological distress.

Background: Low-grade chronic inflammation has been implicated as a risk factor in prostate-related pathologies including benign hyperplasia and cancer. Sweet cherry containing the bioactive anthocyanin (ACN), has demonstrated tumor inhibitory action in model systems, specifically inhibition of inflammatory molecules and prostaglandin biosynthesis. Objective: To assess the urinary and plasma concentrations of ACN from the daily consumption of 3 cups of sweet cherries for 4 weeks and test the relationship of ACN levels and cherry consumption to inflammatory biomarkers in an at risk population. Results: Prostaglandin E2 Metabolite (PGEM) levels were reduced with cherry consumption in men with elevated baseline values. Conclusion: We conclude that 1c (142g) of sweet cherries 3 times daily for 4 weeks significantly reduced the COX-2 metabolite, PGEM, in men with elevated baseline levels. This was the first study to examine the chronic effects of daily sweet cherries on COX-2 inhibition in an at risk population.

Early detection of prostate cancer is problematic due to the lack of a marker that has high diagnostic sensitivity and specificity. The prostate specific antigen test, in combination with digital rectal examination, is the gold standard for prostate cancer diagnosis. However, this modality suffers from low specificity. Therefore, specific markers for clinically relevant prostate cancer are needed. Our objective was to proteomically characterize the conditioned media from human prostate cancer cell lines to identify secreted proteins that could serve as novel prostate cancer biomarkers. An initial proof of principle study of the PC3 prostate cancer cell line was conducted. From this study over 200 proteins were identified in the conditioned media. Through gene ontology analysis and literature searches Mac-2 binding protein was selected as a candidate biomarker for validation in the serum of prostate cancer patients. A preliminarily validation showed that Mac-2 binding protein has discriminatory ability in prostate cancer diagnosis. However, an extended validation did not confirm this. Based on our proof of principle study we optimized our workflow and extended our analysis by culturing three different prostate cell lines [PC3 (bone metastasis), LNCaP (lymph node metastasis), and 22Rv1 (localized to prostate)]. We conducted a bottom-up analysis of each cell line by 2-dimensional liquid chromatography and tandem mass spectrometry. Of the 2124 proteins identified, 12% (329) were classified as extracellular and 18% (504) as membrane-bound. Among the identified proteins were known prostate cancer biomarkers such as PSA and KLK2. To select the most promising candidates for further investigation, tissue specificity, biological function, disease association based on literature searches, and comparison of protein overlap with the proteome of seminal plasma and serum were examined. Based on these results, several candidates were selected for validation in serum of patients with and without prostate cancer. Of these four novel candidates: follistatin, chemokine (C-X-C motif) ligand 16, pentraxin 3 and spondin 2 showed discriminatory ability. Of the four candidates, follistatin was further studied in an extended validation in serum of patients with biopsy confirmed prostate cancer and tissues of prostate cancer patients of low and high grade tumours by immunohistochemistry. In addition, follistatin was also investigated in the tissue of colon and lung cancer where intense staining was observed in one specimen of lung squamous carcinoma.

Prostatacancer är en av de mest vanliga cancersjukdomarna globalt sett och den vanligaste cancerformen i Sverige. Om männen när de får diagnosen är i en relation med någon kan även denna person påverkas av detta. Syftet med denna studie var att undersöka dessa partners erfarenhet av att leva med en man med prostatacancer. En litteraturöversikt baserad på kvalitativt material utfördes. Femton artiklar analyserades och åtta kategorier kunde identifieras: rsn, Anhörigvårdare, Information, Stöd, Sexualitet och Intimitet, Team, Kommunikation och Vardagen. Resultatet visade att kvinnor ofta tar på sig rollen som anhörigvårdare och att de visade ett behov av ytterligare information och stöd. Det sexuella samlivet var även något som kvinnorna ansåg påverkades som en följd av impotensen mannen drabbades av efter behandling. Vissa kvinnor beskrev hur de bemötte cancern tillsammans med sin man som en enhet. En bra kommunikation mellan kvinnorna och deras män uppfattades av vissa kvinnor som viktigt. Många kvinnor beskrev hur deras liv förändrades att de använde sig av olika strategier för att klara av vardagen. När kvinnor antar rollen som anhörigvårdare är det viktigt för dem att de från sjukvården får rätt typ av information och stöd i rätt tidpunkt för att för att kunna förbereda sig för och hantera denna roll. Sjukvården behöver därför utveckla riktlinjer så att kvinnornas behov inkluderas i vårdprocesserna kring mannen. / Prostate cancer is one of the most common cancers globally and the most common cancer in Sweden. If the men when diagnosed are in a relationship with someone, this person may be affected by this. The purpose of this study was to examine the partner's experience of living with a man with prostate cancer. A literature review based on qualitative material was carried out. Fifteen articles were analyzed and eight categories were identified: Changed outlook on life, Caregiver, Information, Support, Sexuality and Intimacy, Team, Communication, and Daily Life. The results showed that women often take on the role of caregivers and that they showed a need for additional information and support. According to women, the impotence the men suffered from after treatment had an impact on their sex-life. Although the women prioritized that their husbands were still alive instead of a functional sex-life many felt that their sex-life had been affected in a negative sense. Some women described how they responded to the cancer with her husband as a unit. Good communication between women and their husbands were perceived by some women as important. Many women described how their life changed and that they used various strategies to cope with everyday life. When women assume the role of caregivers it's important for them to have the right kind of information at the right time given to them by health care providers, this in order to be able to prepare for and respond to the role as a caregiver. Health services therefore need to develop guidelines to ensure that the needs of women are included in the care processes of the men.

The goal of this thesis is to design, implement and evaluate an automated algorithm to detect cancerous tissues and segment the malignant tumour in ultrasound images of the prostate. To accomplish this goal, first, the important image features which would lead to the optimal segmentation are identified. This work focuses on the local texture feature and spatial features. Various approaches to extract the local texture feature are explored, including grey-level co-occurrence matrix (GLCM), recurrent random-pulsed neural networks (RNN), and a novel wavelet-based filter. The spatial features are represented using conventional one dimensional fuzzy membership functions and novel multi-dimensional fuzzy membership functions. The texture and spatial features are combined using the fuzzy inference system. Two of the techniques investigated in this thesis could potentially constitute the basis for key paradigm shifts in medical imaging research. One of these is the idea that medical images in general, and ultrasound images in particular, contain information which are hidden from medical professionals due to limitations in the human visual system. This thesis shows that this information could be extracted using a computerized approach by separating the deterministic components in the image from the indeterministic components, or noise. The other idea concerns the representation of multidimensional statistical distribution information with fuzzy membership functions with the corresponding dimensions. This thesis shows that increasing the number of dimensions with which to represent the statistical distributions results in a more accurate mapping of information that relates to human anatomy, which is essentially 3D in nature. In the thesis, the natures of the various techniques are explored by testing on synthesized images. Then, these approaches are adapted to the ultrasonic prostate cancer segmentation problem and are evaluated with trans-rectal ultrasound images (TRUS). The segmentation using only texture features yields results with high sensitivity. When the spatial features are incorporated using the fuzzy inference system, the specificity of the diagnosis improves dramatically and the overall classification accuracy is also increased. Clinically, this automated diagnostic system could be used as a decision support tool for radiologists when identifying suspicious regions in the prostate from which to draw biopsy samples. The proposed system improves the consistency of the cancer detection process and could provide savings in both time and cost in the prevention and treatment of prostate cancer.

Background Screening for prostate cancer is debated, there is conflicting data on the net benefit of screening. Men who consider screening need to be informed on the pros and cons. Rehospitalization after surgery can be used as an indicator of general quality of care. For radical prostatectomy, little is known on the readmission rate after surgery. Men diagnosed with low- and intermediate-risk prostate cancer have low prostate-cancer specific mortality. However, adverse effects after curative treatment can be severe and decrease quality of life. Curative treatments for prostate cancer differ mainly in the pattern of adverse effects but detailed analysis of long-term adverse effects is lacking. The aim of this thesis was to assess the perioperative quality of radical prostatectomy and the risk of adverse effects after curative treatment for prostate cancer. Material and Methods In this thesis, data from the National Prostate Cancer Register (NPCR) and other nationwide Swedish registers were used. By use of the Swedish personal identity number, NPCR was cross-linked to other registers creating Prostate Cancer data Base Sweden (PCBaSe), a large dataset for research. Results The proportion of men who had received information on the pros and cons of screening for prostate cancer with PSA testing was low (14%) indicating that the majority of men who were screened did not make an informed decision. The risk of rehospitalization within 90 days after radical prostatectomy was approximately 10% and similar after retropubic and robot-assisted radical prostatectomy. Compared to controls, there was an increased risk of adverse effects after both radiotherapy and radical prostatectomy up to twelve years after treatment and the overall risk was quite similar after retropubic and robot-assisted radical prostatectomy. Conclusion Improved information to men on the pros and cons of PSA screening is warranted. The risk of adverse effects was elevated up to 12 years after curative treatment for prostate cancer. The pattern of adverse effects was different after radiotherapy and radical prostatectomy but quite similar after retropubic and robot-assisted radical prostatectomy.