Prostate cancer (PCa) demonstrates notable disease heterogeneity, with African American men (AAM) experiencing a higher incidence and increased mortality rates compared to their European American (EAM) counterparts. While social determinants of health are key contributors to these differences, underlying biological differences remain important drivers of disease severity and outcomes. In the current work, we aimed to explore the interplay between stress mediated reactive oxygen species and inflammatory signaling in promoting progression of PCa in AAM. The study used a retrospective race-matched Genomic Resource Intelligent Discovery (GRID) database (NCT02609269) (n = 8,626), and the prospective VANDAAM study (NCT02723734) (n = 243) to identify hallmarks of oxidative stress and inflammatory signaling in PCa from AAM. We demonstrate that DNA damage repair (DDR) pathway genes are significantly downregulated in AAM-derived PCa, with sustained γH2AX phosphorylation following radiotherapy confirming impaired DNA damage resolution. Transcriptomic analyses further revealed that mitochondrial electron transport chain (ETC) Complex I and Complex III genes are among the most differentially downregulated in AAM, functionally resulting in elevated basal mitochondrial ROS and enhanced sensitivity to ETC inhibition. These mitochondrial ROS-mediated changes promoted lipid peroxidation, plasma membrane remodeling, and extracellular release of ATP and its immunosuppressive metabolite adenosine, selectively in AAM-derived cells and patient-derived explants. Reduced ETC expression further correlated with upregulation of immunosuppressive gene signatures. These findings establish a novel mechanistic framework linking impaired DDR to mitochondrial ETC dysregulation, elevated ROS, and immunosuppressive signature in PCa from AAM, highlighting potential therapeutic targets to address racial disparities in this high-risk population.

Supplementary Figure from Characterization of Epigenomic Alterations in HPV16+ Head and Neck Squamous Cell Carcinomas

Background and objective: Aminopeptidase N (ANPEP) is linked to malignancy in certain tumor types, but its role in aggressive prostate cancer (PCa) is less well defined. Our aim was to characterize ANPEP expression in various PCa stages to determine whether it is a robust prognostic biomarker of aggressive disease. Methods: We established baseline ANPEP expression in benign prostate tissue using multiple large databases. Next, we determined the association between ANPEP expression and various clinicopathologic features and molecular subtypes using ∼170 000 tumor samples from the GRID registry. We calculated median expression values, and reported standardized mean differences. We used receiver operating characteristic and Cox regression analyses to evaluate the diagnostic and prognostic significance of ANPEP for several endpoints, and performed preranked gene set enrichment analysis (GSEA) to identify biological pathways over-represented by race or ANPEP category according to hallmark gene sets. Key findings and limitations: ANPEP expression was higher in normal prostate tissues than in prostate tumors. Advanced clinical stage, higher National Comprehensive Cancer Network risk category, and worse Gleason grade group were all associated with lower median ANPEP expression. Genomic markers of aggressive PCa, such as high Decipher scores, low androgen receptor (AR) activity, ERG overexpression, and loss of PTEN expression, were correlated with lower ANPEP expression. Among patients with locally advanced or metastatic PCa, higher ANPEP expression was significantly associated with more favorable PCa-specific outcomes, including biochemical recurrence, distant metastasis, castration-resistant PCa, and overall survival. GSEA revealed AR upregulation for the ANPEP-high group and men with genomic-derived African race. Conversely, the G2-M DNA damage checkpoint and MYC target genes were enriched in the ANPEP-low and genomic-derived European race groups. Conclusions and clinical implications: Our findings show that ANPEP downregulation is linked to a more aggressive PCa phenotype. Higher ANPEP levels were associated with more favorable outcomes, thereby, establishing ANPEP expression as a prognostic factor for treatment response. Patient summary: We looked at levels of a protein called aminopeptidase N (ANPEP) in prostate tumors using information from large databases. We found that ANPEP is linked to markers that indicate more aggressive disease and that higher ANPEP levels are associated with more favorable treatment outcomes.

Background:Traditional ultrasonography-based prostate biopsy uses a transrectal approach for systematic sampling of 12 cores. The magnetic resonance imaging (MRI) fusion biopsy uses a targeted approach, first identifying regions of interest (ROI) clinically suspicious for prostate cancer (PCa) through MRI, before performing a prostate biopsy aided by ultrasonography. Methods:The single-center institutional retrospective cohort study used 442 men who were recommended for localized PCa management. Cohort A (n = 346) comprised patients who underwent MRI-guided TRUS biopsies, which included both standard 12-core TRUS biopsies and MRI-targeted biopsies performed simultaneously. Cohort B (n = 96) comprised patients who received only standard TRUS biopsy. The primary endpoint was Gleason reclassification, defined as the change in Gleason scores between standard TRUS and targeted region-of-interest (ROI) biopsies among cohort A. Secondary endpoint assessed the role of ROI biopsies in mitigating overtreatment by analyzing the probability of undergoing treatment and the duration of active surveillance (AS). Results:Among men classified as no tumor on standard biopsy, 16.9 % showed Gleason disease on subsequent ROI biopsy. Additionally, ROI group also had a longer time to receive primary treatment (P = .017), as they were more likely to opt for AS (54 %). Lastly, median time spent on AS was longer for the ROI group compared with the non-ROI cohort (P = .002). Conclusion:Adding multiparametric MRI (mpMRI) biopsy to standard TRUS biopsy may increase the detection of PCa. Additionally, mpMRI may allow patients to remain safely on AS, thereby reducing the need of prostate biopsies and improving cost-effectiveness.

The relationship between epigenetic aging and cancer mortality has been well established; however, research is limited on how accelerated epigenetic age may be associated with other cancer-related health outcomes. Epigenetic clocks such as DNAm PhenoAge have recently emerged as biomarkers that can reliably predict morbidity and mortality by assessing DNA methylation at specific gene loci. When estimated epigenetic age surpasses chronological age, the discrepancy is termed epigenetic age acceleration (EAA). Various studies have linked higher EAA to risk factors such as low socioeconomic status, adverse childhood experiences, chronic stress, and HIV infection. Given that many of these factors are also associated with disparities in cancer-related health outcomes, investigating the relationship between accelerated aging and cancer-related symptoms could provide insight into the mechanisms that drive these differences. This study examined correlations between EAA and patient-reported outcomes (PROs) in cancer patients. The analytic data set was obtained by cross-referencing patient data collected as part of two concurrent studies conducted at Moffitt Cancer Center. In one study, cancer patients with and without HIV provided blood samples which were assayed using the Illumina MethylationEPIC BeadChip and translated through the EstimAge website to determine EAA via DNAm PhenoAge and PhenoAge acceleration. In the second study, a larger cohort of cancer patients completed the Edmonton Symptom Assessment Scale (ESAS), which asked them to rate the severity of 12 symptoms on a scale from 0 to 10. A total composite ESAS score was summed to represent overall symptom burden. Patients who provided both an assayed blood sample and a completed ESAS survey were included in this investigation. Spearman’s rank correlation coefficients were calculated to assess the association between PhenoAge acceleration and ESAS symptom severity. Participants included in this study (n=22) were 77% male, 82% White, 9% Black, 4.5% Hispanic or Latino, and 36% HIV-positive. The three most prevalent primary cancers among participants were anal (32%, n=7), lung (14%, n=3), and pancreatic (14%, n=3). The median interval between ESAS completion and blood collection was 70 days (IQR=109). The average composite ESAS score was 35, and the average PhenoAge acceleration was 3.3 years. We observed a moderate correlation between PhenoAge acceleration and several symptoms that met the p<.05 threshold for statistical significance. Specifically, PhenoAge acceleration was moderately correlated with severity of drowsiness (ρ=.60, p<.01), reduced overall well-being (ρ=.49, p=.02), constipation (ρ=.47, p=.03), nausea (ρ=.44, p=.04), and shortness of breath (ρ=.44, p=.04). PhenoAge acceleration was also moderately associated with higher overall symptom burden (ρ=.45, p=.03). We observed that EAA in cancer patients, including those with a comorbid diagnosis of HIV, was associated with worse patient-reported outcomes across a range of symptoms. Sonia T. Brickey, KD L. Jacobs, Aasha I. Hoogland, Ryan M. Putney, Kristina E. Bowles, Heather Jim, Brian D. Gonzalez, Anna E. Coghill. Associations between accelerated epigenetic age and patient-reported outcomes in cancer patients [abstract]. In: Proceedings of the 18th AACR Conference on the Science of Cancer Health Disparities; 2025 Sep 18-21; Baltimore, MD. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2025;34(9 Suppl):Abstract nr C057.

While cancer immunotherapies have revolutionized treatment, their efficacy remains limited for most patients, highlighting tumors' capacity to evade immune detection. Epigenetic modifications, particularly DNA methylation, play a pivotal role in silencing immune-related pathways, thereby facilitating immune escape. By modulating gene expression, methylation disrupts immune activation, contributing to reduced tumor immunogenicity. This study examines the methylation landscape of co-stimulatory and immune checkpoint genes across various cancers to elucidate how these changes influence immune responses. Using The Cancer Genome Atlas (TCGA), we analyzed methylation and gene expression profiles across diverse cancer types. Data from 8,186 solid tumors and 745 adjacent normal tissues were processed via t-distributed stochastic neighbor embedding (t-SNE) with 247 probes targeting 20 immune-related genes. Principal component analysis (PCA) was applied to capture methylation variability across 8,931 tumor and normal samples. To explore the effects of methylation on immune gene expression, we analyzed data from 26 epithelial cancer cell lines treated with 5-azacitidine, a demethylating agent, using the GSE57342 dataset. Post-treatment changes in gene expression were assessed, and survival analyses from prior studies were used to explore clinical correlations. We identified distinct methylation patterns in immune synapse genes, differentiating tumors from adjacent normal tissues. Co-stimulatory genes such as CD40 were hypermethylated in tumors, correlating with decreased expression, whereas immune checkpoint genes like PDL1 and HHLA2 were hypomethylated, correlating with increased expression. This inverse relationship between methylation and gene expression underscores the transcriptional regulatory role of DNA methylation. Treatment with 5-azacitidine reversed hypermethylation in co-stimulatory genes, including CD40, potentially enhancing immune recognition. Methylation levels of these genes were also associated with T cell infiltration into the tumor microenvironment and predicted better outcomes in melanoma patients, underscoring their clinical relevance. This study demonstrates that DNA methylation of immune synapse genes contributes to tumor immune evasion by modulating immune gene expression. Targeting these epigenetic alterations represents a promising strategy to restore tumor immunogenicity and improve immunotherapy outcomes, particularly when combined with immune checkpoint inhibitors. Reversing methylation-driven immune suppression could strengthen the immune response and enhance clinical outcomes for cancer patients. Imene Hamaidi, Anders Berglund, Matthew Mills, Ryan M. Putney, James J. Mulé, Sungjune Kim. DNA methylation of immune synapse genes drives tumor immune evasion [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 4831.

e17052 Background: In the era of precision oncology, the development of reliable prognostic biomarkers is essential to facilitate the detection of aggressive prostate cancer (PCa), especially among African Americans (AA), who endure the disproportionate burden of the disease. Our previous work indicated a significantly higher Aminopeptidase N (ANPEP) expression among AA compared to European American men (EA). While ANPEP plays regulatory and/or modulatory functions in many immune and metabolic pathological conditions, its prognostic role in aggressive prostate tumors remains uninvestigated. Methods: We first evaluated ANPEP expression in prostate tumors and adjacent normal tissue. We then measured the association between changes in normalized ANPEP expression and disease progression by examining correlations with certain clinicopathologic features and genomic signature-defined subgroups from the GRID database. Median (M) ANPEP expression values were calculated, and standardized mean differences (SMDs) were used to assess significant changes in expression levels. SMDs were generally interpreted as small (0.2), medium (0.5), or large (0.8). Finally, we explored the prognostic significance of changes in ANPEP expression by analyzing endpoints such as overall survival (OS), distant metastases (DM), and castration-resistant prostate cancer (CRPC). Results: Normal prostate tissue had higher ANPEP expression compared to prostate tumors (M= 5.64 vs. 4.78, p = <0.001). Both physician-reported and genomic-derived race signatures showed high ANPEP expression in AA (M= 1.30 vs. 0.40, SMD = 0.59) compared to other race groups. Aggressive PCa, as indicated by a higher clinical stage [T1 to T4 (M= 1.21 vs. 0.45, SMD = 0.35)], elevated NCCN risk [low to very high (M= 1.35 vs. 0.46, SMD = 0.29)], and higher pathological stage [pT2 to pT4 (M= 0.86 vs. 0.38, SMD = 0.30)] were associated with an overall decrease in ANPEP expression. Additionally, an increase in adverse pathological features [0 to 3 (M= 0.70 vs. 0.11, SMD = 0.38)] showed a continuous decline in ANPEP expression. Genomic markers of aggressive disease, such as high Decipher score [low vs. high (M= 1.53 vs. 0.59, SMD = 0.60)], low AR [high vs. low (M= 1.26 vs. 0.51, SMD = 0.95)], ERG expression [negative vs. positive (M= 1.11 vs. 0.11, SMD = 1.11)], and PTEN loss [no vs. yes (M= 1.26 vs. 0.22, SMD = 1.04)] showed an overall decrease in ANPEP expression. Lastly, genomic information derived from patients on a number of clinical trials showed that a decrease in ANPEP expression of 0.5 was associated with significant detriment in PCa-specific outcomes, including CRPC [aHR=1.17 (95% CI = 1.04 – 1.31, p =0.007)], DM [aHR=2.27 (95% CI = 1.54 – 3.33, p =<0.001)], and OS [aHR=1.21 (95% CI = 1.06 – 1.39, p =0.004)]. Conclusions: Our study is the first to establish ANPEP as a prognostic biomarker for aggressive PCa using both large prospective genomic databases and clinical trial bio-specimen data.

Supplemental Figure 1 shows the number of non-zero coefficients per variable that was examined in the variable selection processing for the final biomarker panel.

BACKGROUND:Cancer-related deaths for people with human immunodeficiency virus (PWH) are increasing due to longer life expectancies and disparately poor cancer-related outcomes. We hypothesize that advanced biological aging contributes to cancer-related morbidity and mortality for PWH and cancer. We sought to determine the impact of clonal hematopoiesis (CH) on cancer disparities in PWH. METHODS:We conducted a retrospective study to compare the prevalence and clinical outcomes of CH in PWH and people without HIV (PWoH) and cancer. Included in the study were PWH and similar PWoH based on tumor site, age, tumor sequence, and cancer treatment status. Biological aging was also measured using epigenetic methylation clocks. RESULTS:In 136 patients with cancer, PWH had twice the prevalence of CH compared to similar PWoH (23% vs 11%, P = .07). After adjusting for patient characteristics, PWH were 4 times more likely than PWoH to have CH (odds ratio, 4.1 [95% confidence interval, 1.3-13.9]; P = .02). The effect of CH on survival was most pronounced in PWH, who had a 5-year survival rate of 38% if they had CH (vs 59% if no CH), compared to PWoH who had a 5-year survival rate of 75% if they had CH (vs 83% if no CH). CONCLUSIONS:This study provides the first evidence that PWH may have a higher prevalence of CH than PWoH with the same cancers. CH may be an independent biological aging risk factor contributing to inferior survival for PWH and cancer.

Introduction Men with African ancestry have the highest incidence and mortality rates of prostate cancer (PCa) worldwide. Methods This study aimed to identify differentially methylated genes between tumor vs. adjacent normal and aggressive vs. indolent PCa in 121 African American patients. Epigenome-wide DNA methylation patterns in tumor DNA were assessed using the human Illumina Methylation EPIC V1 array. Results Around 5,139 differentially methylated CpG-sites (q < 0.01, l Delta beta l > 0.2) were identified when comparing normal vs. tumor, with an overall trend of hypermethylation in prostate tumors. Multiple representative differentially methylated regions (DMRs), including immune-related genes, such as CD40, Galectin3, OX40L, and STING, were detected in prostate tumors when compared to adjacent normal tissues. Based on an epigenetic clock model, we observed that tumors' total number of stem cell divisions and the stem cell division rate were significantly higher than adjacent normal tissues. Regarding PCa aggressiveness, 2,061 differentially methylated CpG-sites (q < 0.05, l Delta beta l > .05) were identified when the grade group (GG)1 was compared with GG4/5. Among these 2,061 CpG sites, 155 probes were consistently significant in more than one comparison. Among these genes, several immune system genes, such as COL18A1, S100A2, ITGA4, HLA-C, and ADCYAP1, have previously been linked to tumor progression in PCa. Conclusion Several differentially methylated genes involved in immune-oncologic pathways associated with disease risk or aggressiveness were identified. In addition, 261 African American-specific differentially methylated genes related to the risk of PCa were identified. These results can shedlight on potential mechanisms contributing to PCa disparities in the African American Population.

Abstract In the year 2023, prostate cancer (PCa) contributed to 34,700 cancer-related fatalities within the United States. Notably, African American men (AAM) experience disproportionately higher rates of both PCa incidence and mortality compared to their European American counterparts (EAM). To investigate some of the biological factors which contribute to these disparities, we carried out an unbiased genomic expression analysis of PCa stratified by ancestry using retrospective and prospective clinical cohorts. In these analyses, we identified aminopeptidase N (ANPEP, APN, CD13), as the most differentially expressed gene in both self-identified and ancestry derived AAM compared with EAM. Previous studies demonstrated that the aminopeptidase ANPEP cleaves N termini of numerous hormones, cytokines, and chemokines involved in oncogenic signaling, cancer relapse and Inflammation. Specifically, ANPEP trims peptides with an N-terminal L-alanine and (L-cysteinylglycine)-S-conjugates while releasing alanine and glycine, respectively. While the role of ANPEP in cytokine activation is well studied, the contribution of ANPEP to amino acids homeostasis remains unknown. Herein, we aim to characterize the metabolic function of ANPEP and whether ANPEP regulates therapeutic efficacy. Using computational analyses, we demonstrate that expression of ANPEP predominantly correlates with various amino acid transporters. By employing untargeted metabolomics, we subsequently revealed a significant enrichment of one-carbon metabolism in cells that overexpress ANPEP. The one-carbon metabolism connects a series of metabolic pathways including the methionine cycle which in turn regulates methylation of various cellular substrates. We then performed genome-wide methylation profiling demonstrates that ANPEP overexpression is associated with elevated methylation of DNA. Because elevated methylation of DNA which is a determinant of therapeutic efficacy in prostate cancer, future studies will assess whether ANPEP regulation of methylation capacity impacts therapeutic efficacy and disease progression. To clinically validate our findings, we them carried out a fluxomic approach in ex vivo human tumor slice culture from 20 patients of prostate cancer. These studies demonstrate that PCa from AAM exhibit higher demand for methionine and cystine highlighting dominance of one-carbon metabolism. In conclusion, we discovered a new function of ANPEP in regulating one-carbon metabolism in prostate cancer. Citation Format: Asmaa El-Kenawi, Ryan Putney, Anders Berglund, Kosj Yamoah. ANPEP regulates one-carbon metabolism in prostate cancer [abstract]. In: Proceedings of the 17th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2024 Sep 21-24; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2024;33(9 Suppl):Abstract nr C108.

Abstract Disclosure: A. El-Kenawi: None. R. Putney: None. A. Berglund: None. K. Yamoah: None. African American men (AAM) experience disproportionately higher rates of both prostate cancer (PCa) incidence and mortality compared to their European American counterparts (EAM). To investigate some of the biological factors which contribute to these disparities, we carried out an unbiased genomic expression analysis of PCa by ancestry using the matched GRID and VANDAAM cohorts. In this analysis, we discovered aminopeptidase N (ANPEP, APN, CD13), to be the most differentially expressed functional gene in both self-identified and ancestry derived AAM compared with EAM. Further computational analyses revealed that ANPEP correlates with signatures of cholesterol transport, estrogen and androgen receptor (AR) signaling. The functional role of ANPEP in regulating cholesterol metabolism and AR signaling remains unclear. Further computational analyses demonstrate that expression of ANPEP predominantly correlates with various amino acid transporters. In addition, comprehensive metabolomic characterization revealed that ANPEP-overexpressing cells exhibit altered level of one-carbon metabolism metabolites. The one-carbon metabolic pathway regulates amino acids homeostasis and epigenetic mechanisms. Metabolomics-based approach to assess the consumption of over 15 amino acids in patient-derived explants validate enrichment of one-carbon metabolism circuits in AAM. Thus, we further investigated whether ANPEP impact high methylation capacity by performing genome-wide methylation profiling in cells. We unraveled that ANPEP overexpression is associated with elevated methylation of DNA. Our future studies will assess whether ANPEP regulation of methylation capacity have an impact on AR binding to DNA. Presentation: 6/3/2024

Abstract As oropharyngeal cancer (OPC) associated with human papillomavirus (HPV) increases in men, the need for a screening test to diagnose OPC early is crucial. This study agnostically identified differentially methylated CpG sites to identify additional biomarkers to improve screening for early OPC. DNA was extracted from oral gargles of 89 early cases and 108 frequency matched healthy controls, and processed for genome-wide methylation using the Illumina Infinium MethylationEPIC BeadChip. Selected sites were combined with our prior methylation data in the EPB41L3 gene (CpG sites 438, 427, and 425) and oral HPV16 and HPV18 status were considered as binary variables (positive/negative). Lasso regression identified CpG sites strongly associated with early OPC. ROC curves with AUC were generated. The panel was validated utilizing bootstrap resampling. Machine learning analyses identified 14 markers that are significantly associated with early OPC, including one EPB41L3 CpG site (438) and oral HPV16 status. A final model was trained on all available samples using the discovered panel and was able to predict early OPC compared with controls with an AUC of 0.970 on the training set. In the bootstrap validation sets, the average AUC was 0.935, indicating adequate internal validity. Our data suggest that this panel can detect OPC early, however external validation of this panel is needed. Further refinement of a panel of biomarkers to diagnose OPC earlier is urgently needed to prevent complex treatment of OPC and associated comorbidities, while reducing risk of recurrence. Prevention Relevance: This study identified biomarkers using genome-wide methylation to create a panel capable of discerning early oropharyngeal cancer (OPC) from those without OPC. Such a biomarker panel would be an effective tool to detect OPC early and prevent complications of treatment associated with later diagnosis.

Supplementary Table S2. The number and overlap of discordant loci (|Δβ|>0.3) across internal replicates of the Restore and REPLI-g Ligase methods for each patient sample.

While STING-activating agents have shown limited efficacy in early-phase clinical trials, multiple lines of evidence suggest the importance of tumor cell-intrinsic STING function in mediating antitumor immune responses. Although STING signaling is impaired in human melanoma, its restoration through epigenetic reprogramming can augment its antigenicity and T cell recognition. In this study, we show that reversal of methylation silencing of STING in murine melanoma cell lines using a clinically available DNA methylation inhibitor can improve agonist-induced STING activation and type-I IFN induction, which, in tumor-bearing mice, can induce tumor regression through a CD8(+) T cell-dependent immune response. These findings not only provide mechanistic insight into how STING signaling dysfunction in tumor cells can contribute to impaired responses to STING agonist therapy, but also suggest that pharmacological restoration of STING signaling through epigenetic reprogramming might improve the therapeutic efficacy of STING agonists.

Supplemental Materials Methods with additional references.

Supplementary Figure Legend from Expanding Epigenomics to Archived FFPE Tissues: An Evaluation of DNA Repair Methodologies

Abstract Epigenetic modification, involving changes in the methylation of genes, play a key role in the molecular changes seen in many cancers, with prostate cancer (PCa) being one of them. These changes include both hyper-methylation, where genes are silenced and hypo-methylation where genes are over-expressed in tumors. Changes of methylation have also been linked to both drug-resistance and immune-evasion. We will in this work present a visualization suite that we have developed in our analysis of an in-house cohort of adjacent normal and prostate tumor tissue samples from men with African American ancestry (AA). We demonstrate significant changes in immune synapse genes in PCa using a principal component model that describe the dysregulation of both co-stimulatory genes and immune checkpoint genes. We also demonstrate significant changes in STING and changes in immune cell content between adjacent normal and tumor tissue. Treg and B memory cells are increase in tumor tissue, while there is a lower amount of naïve CD4 T cells and naïve B cells. Furthermore, we show that hyper-methylation occurs in many PCa samples, with multiple immune genes being silenced. The degree of hyper-methylation is shown on both a global and a focal view, with Chromosome 17 being most affected by hyper-methylation and zooming in at a region including RARA, IGFBP4, STAT5A, CAVIN1, PLEKHH3, RND2;VAT1, TMEM106A, and DHX8;ETV4 genes. We also compare our results from African American patients to a European cohort, demonstrating an overall similarity in methylation changes but also identifies unique methylation of genes, including GLRX, IFFO1, RASSF1, GEFT that are hyper-methylated in our PCa AA cohort. Additionally, we identify different categories of genes based on their methylation changes across PCa AA samples with different Gleason score. Using a Sankey graph, we classify significant CpG-probes into 8 different categories, describing both hyper- and hypo-methylation and if the changes are seen as a continues change between the different grades area more threshold like. Key genes include SEPT9, CCND2, TMHLE, and SPARCL1 being hyper methylation in tumors with a higher Gleason score, while CDH4, PRDM16, CSMD1, and IFI16 show hypo-methylation in higher Gleason score tumors. We also see an increase of Treg immune cells in PCa samples with high Gleason score. In conclusion, the visualization presented in this study provides an effective way to better understand the epigenetic changes seen in PCa AA men and can potentially also lead to a better understanding of the mechanisms contributing to PCa disparities in the AA population. Citation Format: Anders Berglund, Ryan M. Putney, Kosj Yamoah, Hyun Park, Jong Y. Park. Effective visualization of methylation changes in prostate cancer among African American men [abstract]. In: Proceedings of the 16th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2023 Sep 29-Oct 2;Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2023;32(12 Suppl):Abstract nr C001.