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.

Prostate cancer is the most common cancer affecting men today. Therefore, there is a strong need for accurate biomarkers and successful therapeutic treatments. A novel approach combining a computationally built protein-protein interaction network of proven microRNA protein targets with high throughput proteomics identified ErbB2 and ErbB3 as key proteins in prostate cancer. These results coupled with microRNA array screening of an androgen-independent prostate cancer progression model, substantiated by single microRNA analysis, suggested miR125b as a key tumor suppressor contributing to prostate cancer progression. miR125b expression was shown to be substantially increased in the non-tumorigenic P69 cell line compared to its highly tumorigenic, metastatic M12 variant. Luciferase reporter gene assays including the entire 3’UTR of either ErbB2 or ErbB3 revealed a 2.8- and 2.4-fold decrease (respectively) compared to control vector. Thus, this combinatorial approach has suggested an additional microRNA and its target involved in prostate tumor progression.

Prostate cancer is the second most common cancer in the United States that affects men today. To better treat this disease accurate biomarkers and successful therapeutic treatments are needed. A novel approach to understand the mechanisms behind prostate cancer tumor formation lies in identifying dysregulated micro-RNAs (miRNAs), which are a class of small (18-24 nucleotides) non-coding RNAs that regulate gene expression posttranscriptionally by either inhibiting protein synthesis or signaling messenger-RNA for degradation. Multiple miRNAs were discovered in our highly tumorigenic and metastatic prostate cancer progression model M12 cell line compared to its weakly tumorigenic P69 parental cell line. Various analyses such as human panel analyses, single-miR analyses and patient tumor biopsy samples were analyzed to determine dysregulated miRNAs that contributed to the progression and metastasis of prostate cancer. Together with performing experiments to identify miRNAs, a de novo next generation sequencing approach was applied to identify miRNAs naturally present in biological fluids of normal and healthy subjects. Since, these miRNAs are highly dysregulated in many diseases, including cancer, they can act as potential biomarkers or therapeutic targets to improve treatments for prostate cancer. Essential miRNAs studied for this research were miR-17-3p that is known to target the ErbB2 mRNA; miR-299-5p that directly targets osteopontin (OPN) mRNA, and miR-147b that directly targets many mRNAs, such as COL4A2, ALDH5A1, NDUFA4, SDHD, and IER5. A wide range of miRNAs were identified in six biological fluids: venous blood, menstrual blood, vaginal fluid, semen, saliva, and feces. There were some miRNAs that were common to all 6 body fluids, some unique to each body fluid, and some miRNAs that literature suggested could potentially be biomarkers or normalizers for body fluid characterization.

In radiation therapy, interfraction organ motion introduces a level of geometric uncertainty into the planning process. Plans, which are typically based upon a single instance of anatomy, must be robust against daily anatomical variations. For this problem, a model of the magnitude, direction, and likelihood of deformation is useful. In this thesis, principal component analysis (PCA) is used to statistically model the 3D organ motion for 19 prostate cancer patients, each with 8-13 fractional computed tomography (CT) images. Deformable image registration and the resultant displacement vector fields (DVFs) are used to quantify the interfraction systematic and random motion. By applying the PCA technique to the random DVFs, principal modes of random tissue deformation were determined for each patient, and a method for sampling synthetic random DVFs was developed. The PCA model was then extended to describe the principal modes of systematic and random organ motion for the population of patients. A leave-one-out study tested both the systematic and random motion model’s ability to represent PCA training set DVFs. The random and systematic DVF PCA models allowed the reconstruction of these data with absolute mean errors between 0.5-0.9 mm and 1-2 mm, respectively. To the best of the author’s knowledge, this study is the first successful effort to build a fully 3D statistical PCA model of systematic tissue deformation in a population of patients. By sampling synthetic systematic and random errors, organ occupancy maps were created for bony and prostate-centroid patient setup processes. By thresholding these maps, PCA-based planning target volume (PTV) was created and tested against conventional margin recipes (van Herk for bony alignment and 5 mm fixed [3 mm posterior] margin for centroid alignment) in a virtual clinical trial for low-risk prostate cancer. Deformably accumulated delivered dose served as a surrogate for clinical outcome. For the bony landmark setup subtrial, the PCA PTV significantly (p30, D20, and D5 to bladder and D50 to rectum, while increasing rectal D20 and D5. For the centroid-aligned setup, the PCA PTV significantly reduced all bladder DVH metrics and trended to lower rectal toxicity metrics. All PTVs covered the prostate with the prescription dose.

The chemokine receptor CCR5 (CCR5) plays an integral role within the inflammatory network of cells. Importantly, CCR5 is a mediator in several disease states and can be targeted using small molecule antagonists. Within this work, CCR5’s role in prostate cancer and HIV/AIDS has been exploited in order to develop potential therapeutics and probes. First, a series of novel compounds was designed by using pharmacophore-based drug design based upon known CCR5 antagonists and molecular modeling studies of the CCR5 receptor’s three-dimensional conformation. Once synthesized, these compounds were tested for their CCR5 antagonism and their anti-proliferative effects in several prostate cancer cell lines. The data from both the calcium mobilization studies and the anti-proliferation studies suggests that the compounds synthesized have activity as CCR5 antagonists and as anti-proliferative agents in certain prostate cancer cell lines. In addition, a bivalent ligand containing both a mu opioid receptor (MOR) and a CCR5 antagonist pharmacophore was designed and synthesized in order to study the pharmacological profile of the putative CCR5-MOR heterodimer and its relation with NeuroAIDS. The structural-activity relationship between the bivalent ligand and the heterodimer was studied with radio-ligand binding assays, functional assays, HIV-1 fusion assays, cell fusion assays, and in silico molecular dynamics. The subsequent bivalent ligand was proven to be a potent inhibitor in both an artificial cell fusion assay mimicking HIV invasion and a native HIV-1 invasion assay using live virus. In all, two novel sets of compounds were synthesized that targeted either CCR5 or the CCR5-MOR heterodimer. For the CCR5 antagonists, as leads for prostate cancer therapeutics, further work needs to be done to ascertain and develop their structure-activity-relationship. This library of novel compounds was shown as promising leads as CCR5 and anti-prostate cancer agents. The bivalent ligand targeting the CCR5-MOR heterodimer proved to be a potent and tissue-specific inhibitor for neuroAIDS where the known treatment, maraviroc, is less efficacious and fails to inhibit virus entry in the presence of morphine. Both projects illustrate the roles that CCR5 plays in these two unique diseases.

This is a compilation of 3 abstracts for the three manuscripts included in this dissertation. I. Meta-Analysis: Racial Disparities in Prostate Cancer Survival: Prostate cancer is the second leading cause of cancer-related mortality in men. Previous studies have drawn inconsistent conclusions on racial differences in prostate cancer survival. This meta-analysis was conducted to investigate the relationship between race and survival from prostate cancer. A systematic review of published articles from 1968 to 2007 assessing survival from prostate cancer among African American and White men was conducted. The search yielded 20 eligible published manuscripts. Analysis of unadjusted studies showed African American men have an increased risk of all-cause mortality (Hazard ratio (HR) = 1.47, 95% confidence interval (CI): 1.31, 1.65, P < 0.001). However, examination of adjusted studies showed no difference (HR = 1.07, 95% CI: 0.94, 1.22, P = 0.308). No statistically significant difference was observed in prostate cancer-specific survival in both analyses using unadjusted (HR = 1.11, 95% CI: 0.94, 1.31, P = 0.209) and adjusted studies (HR = 1.15, 95% CI: 0.95, 1.41, P = 0.157). There was evidence of heterogeneity that was unexplained by factors analyzed in overall survival but explained by stage in prostate cancer-specific survival. This meta-analysis concludes that there are no racial differences in the overall and prostate cancer-specific survival between African American and White men. II. Case-Control study: Association between Family History of Cancers and Prostate Cancer: Family history of prostate cancer is an established risk factor for prostate cancer. However, the relationship between family history of cancers other than prostate cancer and prostate cancer risk is inconclusive. This study sought to examine the association between family history of cancers and prostate cancer. A case-control study was conducted in which cases and controls were randomly selected from a large urology clinic in Central Virginia. Cases were 600 histologically confirmed prostate cancer patients who were diagnosed between January 2000 and December 2005, and controls were 686 patients who visited the clinic during the same period and diagnosed with urological illnesses other than cancers and prostate-related problems. Data on family history of cancers, lifestyle and demographic factors were collected. Unconditional logistic regression analysis was used to estimate the odds ratios and the corresponding 95% confidence intervals after adjustment for potential confounding factors. Multiple comparisons adjustments were made using Bonferroni adjustment. Men with family history of any cancer in first-degree relatives including parents (OR=2.42, 95% CI: 1.53, 3.84) and parents only (OR=1.90, 95% CI: 1.23, 2.94) were at increased risk of developing prostate cancer compared to men with no such family history of cancer. Significant increased risk was also observed with family history of prostate cancer in first-degree relatives (OR=2.68, 95% CI: 1.53, 4.69) and parents only (OR=3.26, 95% CI: 1.71, 6.24) compared to men with no family history of prostate cancer. Even after adjustments for multiple comparisons, the significance persisted both in first-degree relatives (OR=2.68, 95% CI: 1.16, 6.21) and parents alone (OR=3.26, 95% CI: 1.24, 8.63). No association was found with family history of other cancers including breast, colon, lung, skin, digestive tract, stomach, liver, pancreas, female cancers, urogenital, urinary bladder, brain, blood and lymph node and other cancers and risk of prostate cancer. This study demonstrated an increased prostate cancer risk for men with a family history of any cancer or prostate cancer in first-degree relatives including parents and parents alone. Health care providers need to be aware of the potential risk of family history of cancers on prostate cancer. III. Case-Control study: Association between Obesity and Prostate Cancer: Obesity is a major public health problem in the United States. Several studies have investigated the association between obesity and prostate cancer risk. However the impact of early-adult obesity on prostate cancer is not well studied. This study proposes to investigate the relationship between prostate cancer and early-onset obesity and current obesity. A case-control study was conducted to investigate the relationship between obesity and prostate cancer in a large urology clinic population in Central Virginia. Cases included histologically confirmed prostate cancer patients of all stages and grades diagnosed from January 2000 to December 2005. Controls were patients who were diagnosed with urological illness other than cancers and prostate-related problems. Self-reported data was collected on anthropometric, lifestyle and demographic factors through a mail survey. Unconditional logistic regression analysis was conducted to investigate the association between prostate cancer and early-onset obesity (BMI at age 18) and current obesity. Odds ratios and corresponding 95% confidence intervals were calculated after accounting for significant interaction terms and adjusting for potential confounding variables. This study showed statistically significant association between BMI at age 18 and prostate cancer risk in the multivariate analysis when BMI was evaluated as a continuous variable. There was a 7% decrease in the odds of prostate cancer risk for every 1 kg/m(2) increment in BMI at age 18 (OR=0.93, 95% CI: 0.87, 0.98). Analysis of BMI at age 18 as a categorical variable also showed reduced risk though statistically non-significant. Obese men (OR=0.62, 95% CI: 0.12, 3.08) and overweight men (OR=0.60, 95% CI: 0.35, 1.05) had a non-significant decreased risk of developing prostate cancer compared to normal weight men at age 18. Examination of current BMI showed a non-statistically significant decreased risk of prostate cancer when examined as a continuous variable. However, there was significant interaction between current BMI treated categorically and age. This study concludes that there is decreased prostate cancer risk associated with increasing BMI at age 18. Future large prospective studies are needed to better understand the association between early-onset obesity and risk of prostate cancer and explore the biological factors associated especially in the early ages. This document was created in Microsoft Word 2003.

Breast and prostate cancer incidence and mortality have been steadily decreasing. Reasons for these reductions may be related to increased rates of cancer screening and other factors such as improvements in diet, including consumption of fruits and vegetables. We wanted to determine if individuals who get screened for breast and prostate cancer are more or less likely to consume adequate servings of fruit and vegetables. A cross-sectional study using the BRFSS survey was conducted. Individuals included in this study (n=26,222), were asked about their breast or prostate cancer screening history. They were also asked about their servings per day of fruit and vegetables. Statistical analyses were conducted using the SAS 9.2 software program. Logistic regression analyses were conducted on the variables and potential confounders. Over 40% of individuals who did not screen for breast and prostate cancer were in the 50-59 years of age category. A trend was seen with younger age groups being less likely to consume 3 or more daily servings of fruit and vegetables than their older counterparts. Another trend was seen in education levels. Individuals with lower education were less likely to consume at least 3 daily servings of fruit and vegetables. There was a statistically significant association between cancer screening and servings of fruit and vegetables per day. Individuals who were screened for either breast or prostate cancer were 52% more likely to consume 3 or more servings of fruit and vegetables than those who did not screen for either breast or prostate cancer (OR=1.52, 95% CI: 1.29-1.79). Further research needs to be conducted related to how other health behaviors may be related to cancer screening adherence and fruit/vegetable intake.