Search

Supplementary Figure 1 from A Novel Prostate Cancer Susceptibility Locus at 19q13

Supplementary Table 1 from A Novel Prostate Cancer Susceptibility Locus at 19q13

Supplementary Figure 1 from Germ-Line Mutation of NKX3.1 Cosegregates with Hereditary Prostate Cancer and Alters the Homeodomain Structure and Function

A two-stage genome-wide association study (GWAS) of the Cancer Genetic Markers of Susceptibility (CGEMS) initiative identified single nucleotide polymorphisms (SNP) in 150 regions across the genome that may be associated with prostate cancer (PCa) risk. We filtered these results to identify 43 independent SNPs where the frequency of the risk allele was consistently higher in cases than in controls in each of the five CGEMS study populations. Genotype information for 22 of these 43 SNPs was obtained either directly by genotyping or indirectly by imputation in our PCa GWAS of 500 cases and 500 controls selected from a population-based case-control study in Sweden [Cancer of the Prostate in Sweden (CAPS)]. Two of these 22 SNPs were significantly associated with PCa risk (P < 0.05). We then genotyped these two SNPs in the remaining cases (n = 2,393) and controls (n = 1,222) from CAPS and found that rs887391 at 19q13 was highly associated with PCa risk (P = 9.4 × 10−4). A similar trend of association was found for this SNP in a case-control study from Johns Hopkins Hospital (JHH), albeit the result was not statistically significant. Altogether, the frequency of the risk allele of rs887391 was consistently higher in cases than controls among each of seven study populations examined, with an overall P = 3.2 × 10−7 from a combined allelic test. A fine-mapping study in a 110-kb region at 19q13 among CAPS and JHH study populations revealed that rs887391 was the most strongly associated SNP in the region. Additional confirmation studies of this region are warranted. [Cancer Res 2009;69(7):2720–3]

NKX3.1, a gene mapped to 8p21, is a member of the NK class of homeodomain proteins and is expressed primarily in the prostate. NKX3.1 exerts a growth-suppressive and differentiating effect on prostate epithelial cells. Because of its known functions and its location within a chromosomal region where evidence for prostate cancer linkage and somatic loss of heterozygosity is found, we hypothesize that sequence variants in the NKX3.1 gene increase prostate cancer risk. To address this, we first resequenced the NKX3.1 gene in 159 probands of hereditary prostate cancer families recruited at Johns Hopkins Hospital; each family has at least three first-degree relatives affected with prostate cancer. Twenty-one germ-line variants were identified in this analysis, including one previously described common nonsynonymous change (R52C), two novel rare nonsynonymous changes (A17T and T164A), and a novel common 18-bp deletion in the promoter. Overall, the germ-line variants were significantly linked to prostate cancer, with a peak heterogeneity logarithm of odds of 2.04 (P = 0.002) at the NKX3.1 gene. The rare nonsynonymous change, T164A, located in the homeobox domain of the gene, segregated with prostate cancer in a family with three affected brothers and one unaffected brother. Importantly, nuclear magnetic resonance solution structure analysis and circular dichroism studies showed this specific mutation to affect the stability of the homeodomain of the NKX3.1 protein and decreased binding to its cognate DNA recognition sequence. These results suggest that germ-line sequence variants in NKX3.1 may play a role in susceptibility to hereditary prostate cancer and underscore a role for NKX3.1 as a prostate cancer gatekeeper. (Cancer Res 2006; 66(1): 69-77)

Supplementary Table 3 from A Novel Prostate Cancer Susceptibility Locus at 19q13

Supplementary Table 2 from A Novel Prostate Cancer Susceptibility Locus at 19q13

Supplementary Table 4 from A Novel Prostate Cancer Susceptibility Locus at 19q13

Although obesity at the time of prostatectomy has been associated with prostate cancer recurrence, it is unknown whether obesity before or after surgery, or weight change from the years prior to surgery to after surgery is associated with recurrence. Thus, we examined the influence of obesity and weight change on recurrence after prostatectomy. We conducted a retrospective cohort study of 1,337 men with clinically localized prostate cancer who underwent prostatectomy performed during 1993–2006 by the same surgeon. Men self-reported weight and physical activity at 5 years before and 1 year after surgery on a survey during follow-up. Mean follow-up was 7.3 years. We estimated multivariable-adjusted HRs of prostate cancer recurrence comparing obesity at 5 years before and at 1 year after surgery with normal weight, and a gain of more than 2.2 kg from 5 years before to 1 year after surgery with stable weight. During 9,797 person years of follow-up, 102 men recurred. Compared with men who had stable weight, those whose weight increased by more than 2.2 kg had twice the recurrence risk (HR = 1.94; 95% CI, 1.14–3.32) after taking into account age, pathologic stage and grade, and other characteristics. The HR of recurrence was 1.20 (95% CI, 0.64–2.23) and 1.72 (95% CI, 0.94–3.14) comparing obesity at 5 years before and at 1 year after surgery, respectively, with normal weight. Physical activity (≥5 h/wk) did not attenuate risk in men who gained more than 2.2 kg. By avoiding weight gain, men with prostate cancer may both prevent recurrence and improve overall well-being. Cancer Prev Res; 4(4); 544–51. ©2011 AACR.

Two single nucleotide polymorphisms (SNP; rs5945572 and rs5945619) at Xp11 were recently implicated in two genome-wide association studies of prostate cancer. Using a family-based association test for these two SNPs in 168 families with prostate cancer, we showed in this study that the risk alleles of the two reported SNPs were overtransmitted to the affected offspring (P= 0.009 for rs5945372 and P = 0.03 for rs5945619), which suggested that the observed association in case-control studies were not driven by potential population stratification. We also did a fine-mapping study in the ∼800 kb region at Xp11 between two independent case-control studies, including 1,527 cases and 482 controls from Johns Hopkins Hospital and 1,172 cases and 1,157 controls from the Prostate, Lung, Colon and Ovarian Cancer screening trial. The strongest association was found with SNPs in the haplotype block in which the two initial reported SNPs were located, although many SNPs in the ∼140 kb region were highly significant in the combined allelic tests (P = 10−5 to 10−6). The second strongest association was observed with SNPs in the ∼286 kb region at another haplotype block (P = 10−4 to 10−5), ∼94 kb centromeric to the first region. The significance of SNPs in the second region decreased considerably after adjusting for SNPs at the first region, although P values remained at

Purpose: Fifteen independent genetic variants have been implicated in prostate cancer risk by recent genome-wide association studies. However, their association with clinicopathologic features of prostate cancer is uncertain. Experimental Design: We systematically evaluated these 15 variants in 1,563 prostate cancer patients undergoing radical prostatectomy, taking advantage of the uniform tumor stage and grade information available for each of these cases. Associations of these variants with aggressiveness, pathologic Gleason scores, pathologic stage, age at diagnosis, or serum prostate-specific antigen (PSA) levels were tested. Results: After adjusting for multiple testing, none of the single nucleotide polymorphisms was individually or cumulatively associated with aggressiveness or individual clinicopathologic variables of prostate cancer such as Gleason scores, pathologic stage, or age at diagnosis of prostate cancer. The reported risk allele (G) for single nucleotide polymorphism rs2735839 in the KLK3 gene at 19q13 was more frequent in less aggressive prostate cancer patients (0.89) than in more aggressive prostate cancer patients (0.86; nominal P = 0.03) or in controls (0.86; nominal P = 0.04). Considering that this allele was also significantly associated with higher serum PSA levels among controls (nominal P = 0.003), the observed trend of higher frequency of this risk allele between less and more aggressive prostate cancer, or between less aggressive and controls may be due to detection bias of PSA screening. Conclusions: Prostate cancer risk variants recently discovered from genome-wide case-control association studies are not associated with clinicopathologic variables in this population. Case-case studies are urgently needed to discover genetic variants that predict tumor aggressiveness.

Prostate cancer is a leading and increasingly prevalent cause of cancer death in men. Whereas family history of disease is one of the strongest prostate cancer risk factors and suggests a hereditary component, the predisposing genetic factors remain unknown. We first showed that KLF6 is a tumor suppressor somatically inactivated in prostate cancer and since then, its functional loss has been further established in prostate cancer cell lines and other human cancers. Wild-type KLF6, but not patient-derived mutants, suppresses cell growth through p53-independent transactivation of p21. Here we show that a germline KLF6 single nucleotide polymorphism, confirmed in a tri-institutional study of 3,411 men, is significantly associated with an increased relative risk of prostate cancer in men, regardless of family history of disease. This prostate cancer–associated allele generates a novel functional SRp40 DNA binding site and increases transcription of three alternatively spliced KLF6 isoforms. The KLF6 variant proteins KLF6-SV1 and KLF6-SV2 are mislocalized to the cytoplasm, antagonize wtKLF6 function, leading to decreased p21 expression and increased cell growth, and are up-regulated in tumor versus normal prostatic tissue. Thus, these results are the first to identify a novel mechanism of self-encoded tumor suppressor gene inactivation and link a relatively common single nucleotide polymorphism to both regulation of alternative splicing and an increased risk in a major human cancer.

Background: Obesity and weight gain are associated with poor prostate cancer outcome; underlying mechanisms are unknown. Obesity has been more strongly associated with lethal disease in men positive for the TMPRSS2:ERG gene fusion than negative for this fusion. PTEN loss is associated with increased risk of lethal progression; its association with obesity has not been explored. We evaluated whether prevalence of ERG expression and PTEN loss in prostate tumors differs by BMI and weight change. Methods: From a retrospective cohort study of 1,337 men with clinically-localized prostate cancer who underwent prostatectomy at Johns Hopkins (1993-2006), we sampled 291 men by combination of BMI at 1 year after surgery (normal, overweight, obese) and weight change from 5 years before to 1 year after surgery (loss, maintenance, gain). Within each category, men were frequency-matched on age, Gleason sum, organ confinement, and surgery year. We used genetically validated immunohistochemistry assays to assess ERG positivity and heterogeneous PTEN loss. Logistic regression was used to estimate odds ratios (ORs) and 95% confidence intervals (CIs) for the association of BMI and weight change (adjusted for height, starting weight) with ERG positivity and heterogeneous PTEN loss. Results: Mean age was 56.1 years and mean Gleason sum was 6.2; 73% had organ-confined disease. Prevalence of ERG positivity did not statistically significantly differ by BMI or weight change. However, when jointly categorized by BMI and weight change, overweight/obese men who maintained (OR=2.5; 95%CI 1.1-5.7) or gained weight (OR=2.5; 95%CI 1.1-5.8), and normal weight men who gained weight (OR=4.7; 95%CI 1.4-15.2) were significantly more likely to have ERG positive tumors compared with normal weight men who maintained weight. Overweight/obese men were also significantly more likely (OR=2.4; 95%CI 1.3-4.7) to have PTEN loss compared with normal weight men. When stratified by ERG status, this association appeared to be stronger for men with ERG positive tumors (OR=2.9; 95%CI 1.3-6.8) than for men with ERG negative tumors (OR=1.6; 95%CI 0.6-4.9). Prevalence of PTEN loss did not differ by weight change. When jointly categorized by BMI and weight change, overweight/obese men who maintained (OR=2.0; 95%CI 0.8-5.3) or gained weight (OR=1.9; 95%CI 0.7-5.1) appeared to be more likely to have PTEN loss compared with normal weight men who maintained weight. Conclusions: Overweight/obese men who maintained or gained weight circa diagnosis and normal weight men who gained weight were more likely to have ERG positive tumors. Overweight/obese men were more likely to have tumors with PTEN loss; this association was stronger among overweight/obese men who also had ERG positive tumors. The TMPRSS2:ERG gene fusion and PTEN loss may contribute to the increased risk of poor prostate cancer outcomes among overweight/obese men. Funding: Prostate Cancer Foundation Citation Format: Janelle S. Ho, Ibrahim Kulac, Tamara L. Lotan, John R. Barber, Patrick C. Walsh, Misop Han, Angelo M. De Marzo, Elizabeth A. Platz, Corinne E. Joshu. ERG expression and PTEN loss by BMI and weight change in men with prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5324. doi:10.1158/1538-7445.AM2017-5324

The loss of cell cycle control is believed to be an important mechanism in the promotion of carcinogenesis. CDKN1B (p27) belongs to the Cip/Kip family and functions as an important cell cycle gatekeeper. Several lines of evidence from clinical studies and laboratory experiments demonstrate that CDKN1B is an important tumor suppressor gene in prostate cancer etiology. In addition, a case-control study has shown that the 326T/G (V109G) polymorphism in CDKN1B is associated with advanced prostate cancer. In light of the evidence for linkage between the chromosomal location of the CDKN1B gene (12p13) and prostate cancer susceptibility in several hereditary prostate cancer (HPC) populations, we hypothesized that sequence variants of CDKN1B play a role in HPC. To test this hypothesis, we first resequenced this gene in 96 HPC probands to identify germ-line mutations and sequence variants. We then genotyped the identified sequence variants among all family members of 188 HPC families and tested for their cosegregation with prostate cancer. In total, 10 sequence variants were identified, including three nonsynonymous changes. A family-based test, which is free from the effects of population stratification, revealed a significant association between single nucleotide polymorphism (SNP) -79C/T and prostate cancer (with a nominal P of 0.0005). The C allele of -79C/T was overtransmitted from parents to their affected offspring. Evidence for this association was primarily contributed by affected offspring whose age at diagnosis was

Background: Although Epstein-Barr virus (EBV), a common gamma herpesvirus known to contribute to several cancers (e.g., Burkitt's lymphoma), has been detected in prostate tissue, results from the few epidemiologic studies of EBV infection and prostate cancer have been null. One possible reason for these null findings may be that not all EBV infections contribute to prostate cancer risk. For instance, for other proposed EBV-associated conditions, such as multiple sclerosis, infections acquired later in life (e.g., in adolescence or early adulthood) when EBV infection tends to manifest as infectious mononucleosis (IM) have been found to be more important for risk than those acquired earlier in life when EBV infection is typically asymptomatic. To begin to explore the importance of later-onset EBV infections for prostate carcinogenesis, we investigated the extent of prostate pathogenesis during adult-onset IM by measuring prostate specific antigen (PSA) concentration as a marker of prostate infection, inflammation, and cell damage in adult-onset IM cases and controls with stored serum in the Department of Defense Serum repository (DoDSR). Methods: IM cases were men diagnosed with IM in 1998–2003 (n=55) and controls were a sample of men with no IM diagnoses from 1995–2006 (n=255). For each IM case, we selected two archived serum specimens from the DoDSR, one collected after their diagnosis (acute specimen, range: 1 day-3 years after diagnosis) and the first specimen collected immediately before their acute specimen (pre-acute specimen, range: 46 days-4 years before diagnosis). We also selected two specimens for each control, one collected from 1998–2003 (“acute”) and the first specimen collected immediately before their “acute” specimen (“pre-acute”). All specimens were anonymized before release from the DoDSR. We measured PSA in each of these stored specimens by the Access Hybritech PSA assay. We compared the absolute and relative change in PSA between the pre-acute and acute specimens for cases and controls using logistic regression and adjusting for age, race, and time between specimens. Results: IM cases were significantly more likely to have a large increase in PSA between their pre-acute and acute specimens than controls. This finding was observed both when PSA increase was defined as a large absolute (≥0.4 ng/mL; 9.9% versus 2.7%, p=0.033) or relative rise (≥40%; 25.2% versus 8.9%, p=0.0021). Similar results were observed when men with any infectious disease or genitourinary diagnoses between their pre-acute and acute specimens were excluded. Conclusions: Our findings suggest that EBV may infect the prostate and contribute to prostate inflammation and/or cell damage in some men with adult-onset IM. However, another interpretation of our findings that also warrants further investigation is the possibility of systemic inflammation-mediated PSA elevation. This alternate interpretation might have implications not only for a role of later-onset EBV infection in prostate carcinogenesis, but also for the specificity of PSA as a screening test for prostate cancer when used below the currently proposed age for screening initiation of 40 years. Funding: This study was funded by the Patrick C. Walsh Prostate Cancer Research Fund. Citation Information: Cancer Prev Res 2011;4(10 Suppl):A96.

Background: Obesity measured at or near time of prostatectomy has been associated with increased risk of prostate cancer recurrence. However, it is unknown whether obesity or weight gain in the years prior to surgery is associated with recurrence. In addition, whether physical inactivity or sedentary behavior exacerbates the obesity-related risk of recurrence has not been studied. Methods: We conducted a retrospective cohort study of men with clinically-localized prostate cancer who underwent radical prostatectomy performed by one surgeon at Johns Hopkins between 1/93 − 3/06 and who previously had not had hormone or radiation therapy. The men were followed for recurrence, defined as PSA recurrence, metastasis, or prostate cancer death. A survey on dietary, lifestyle and medical factors, including weight, height, physical activity, and sedentary behavior 5 years before surgery and 1 year after, was mailed to the men residing in the U.S. as of 11/07. We classified men as normal body mass index (BMI, Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 883.

Elliott et al. (1) report that prostate-specific antigen (PSA) performance was significantly better in identifying high-grade disease in men with smaller prostates. They use this observation to support the hypothesis that the increased detection of high-grade disease in the finasteride arm of the Prostate Cancer Prevention Trial was due to an artifact in which the reduction in prostate size made it possible for PSA testing to identify the cancer better in smaller prostates. In their analysis, however, they do not consider another explanation for their findings. It is well established that patients with smaller prostates have more high-grade disease, more extraprostatic extension with positive surgical margins, and higher rates of biochemical progression following radical prostatectomy than men with larger prostates (2–4). Simply stated, did the authors find more high-grade cancer in smaller prostates because there was more high-grade disease to find and not because PSA was better at finding it? Although PSA may perform better in the absence of prostatic enlargement, when Freedland et al. (2) excluded men with T1c disease and examined only men who had abnormal digital rectal examinations, they found that the association between smaller prostate size, higher grade advanced disease, and biochemical progression remained statistically significant. In the authors9 study, 39% of their patients were also diagnosed with palpable disease and those men with smaller prostates also had higher rates of Gleason 7 to 10 disease and higher PSA density (PSA divided by prostate volume). If so, how does this help explain the Prostate Cancer Prevention Trial findings as an artifact? If high-volume, high-grade disease was detected more often during the first 7 years in the finasteride arm, then one would assume eventually over time that there would be fewer patients with high-grade disease. Instead, at 7 years, there were almost twice as many high-grade cancers in the finasteride arm, and in the end of study biopsies, the number was the same, not fewer (5). The authors are at an institution that has emphasized the value of measuring tumor volume in radical prostatectomy specimens and may be able to determine if the volume of high-grade tumor in the smaller prostates was the same as in larger prostates. This would support their conclusion. If not, how is this an artifact? Disclosure of Potential Conflicts of Interest No potential conflicts of interest were disclosed.