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Introduction Fibroblasts maintain tissue and organ homeostasis through output of extracellular matrix that affects nearby cell signaling within the stroma. Altered fibroblast signaling contributes to many disease states and the extracellular matrix secreted by fibroblasts has been used to stratify patient by outcome, recurrence and therapeutic resistance. Recent advances in imaging mass spectrometry allow access to single cell fibroblasts and their ECM niche within clinically relevant tissue samples. Areas covered We review biological and technical challenges as well as new solutions to proteomic access of fibroblast expression within the complex tissue microenvironment. Review topics cover conventional proteomic methods for single fibroblast analysis and current approaches to accessing single fibroblast proteomes by imaging mass spectrometry approaches. Strategies to target and evaluate the single cell stroma proteome on the basis of cell signaling are presented. Expert opinion The promise of defining proteomic signatures from fibroblasts and their extracellular matrix niches is the discovery of new disease markers and the ability to refine therapeutic treatments. Several imaging mass spectrometry approaches exist to define the fibroblast in the setting of pathological changes from clinically acquired samples. Continued technology advances are needed to access and understand the stromal proteome and apply testing to the clinic.

Inverted Meckel's diverticulum is an entity often discovered incidentally or through a clinical evaluation for gastrointestinal bleeding. While rare, inverted Meckel's diverticulum should be considered in the evaluation of a patient presenting with gastrointestinal bleeding, intestinal obstruction, or intussusception. In this case, a 67-year-old female with a remote history of surgically treated breast cancer presents to an urgent care facility with weakness and fatigue. She was found to be anemic with hemoglobin of 4. Imaging revealed a blind-ending pouch in the mid to distal ileum consistent with an inverted Meckel's diverticulum. Inverted Meckel's diverticulum is identified on computerized tomography as an intraluminal, blind-ending structure in the mid to distal ileum. The possibility of a lead point should be investigated and surgical resection is indicated to prevent intestinal obstruction.

BACKGROUND: The emergence of reactive stroma is a hallmark of PCa progression and a potential source for prognostic and diagnostic markers of PCa. Collagen is a main component of reactive stroma and changes systematically and quantitatively to reflect the course of PCa, yet has remained undefined due to a lack of tools that can define collagen protein structure. Here we use a novel collagen-targeting proteomics approach to investigate zonal regulation of collagen type proteins in PCa prostatectomies. METHODS: Prostatectomies from 9 patients were divided into zones containing 0, 5, 20, 70–80% glandular tissue and 0, 5, 25, 70% by mass of prostate cancer tumor following the McNeal model. Tissue sections from zones were graded by a pathologist for Gleason score, percent tumor present, percent prostatic intraepithelial neoplasia and/or inflammation. High resolution accurate mass (HRAM) collagen targeting proteomics was done on a select subset of tissue sections from patient-matched tumor or nontumor zones. Imaging mass spectrometry was used to investigate collagen type regulation corresponding to pathologist defined regions. RESULTS: Complex collagen proteomes were detected from all zones. COL17A and COL27A increased in zones of inflammation compared to zones with tumor present. COL3A1, COL4A5, and COL8A2 consistently increased in zones with tumor content, independent of tumor size. Collagen hydroxylation of proline (HYP) was altered in tumor zones compared to zones with inflammation and no tumor. COL3A1 and COL5A1 showed significant changes in HYP peptide ratios within tumor compared to zones of inflammation (2.59 ± 0.29 p-value 0.015; 3.75 ± 0.96 p-value 0.036, respectively). By imaging mass spectrometry COL3A1 showed defined localization and regulation to tumor pathology. COL1A1 and COL1A2 showed gradient regulation corresponding to PCa pathology across zones. Pathologist defined tumor regions showed significant increases in COL1A1 HYP modifications compared to COL1A2 HYP modifications. Certain COL1A1 and COL1A2 peptides could discriminate between pathologist defined tumor and inflammatory regions. CONCLUSIONS: Site-specific post-translational regulation of collagen structure by proline hydroxylation may be involved in reactive stroma associated with PCa progression. Translational and post-translational regulation of collagen protein structure has potential for new markers to understand PCa progression and outcomes.

Simple Summary Extracellular matrix within the tumor microenvironment influences signaling, controls molecular diffusion of nutrients and growth factors, alters immunogenicity, and contributes to disease progression and therapeutic response. ECM is secreted by multiple cell types, including tumor cells, fibroblasts, and immune cells, yet there are limited approaches that link the cell type to the ECM proteins within the heterogeneous tumor microenvironment. Here, we show that integrating immunohistochemistry (IHC) with extracellular matrix (ECM) imaging mass spectrometry allows ECM proteomic profiling based on patterns of diverse cell types and proteins in tissue. The developed approach is demonstrated using phosphatase and tensin homolog (PTEN) staining and ECM imaging/proteomics on the same tissue sections in normal breast and in a tissue microarray of breast tumor and normal adjacent tissue. The data suggests that PTEN expression in tumor and in normal adjacent tissue may be associated with different collagen types and regulation by post-translational sites of modification. Abstract Breast stroma plays a significant role in breast cancer risk and progression yet remains poorly understood. In breast stroma, collagen is the most abundantly expressed protein and its increased deposition and alignment contributes to progression and poor prognosis. Collagen post-translation modifications such as hydroxylated-proline (HYP) control deposition and stromal organization. The clinical relevance of collagen HYP site modifications in cancer processes remains undefined due to technical issues accessing collagen from formalin-fixed, paraffin-embedded (FFPE) tissues. We previously developed a targeted approach for investigating collagen and other extracellular matrix proteins from FFPE tissue. Here, we hypothesized that immunohistochemistry staining for fibroblastic markers would not interfere with targeted detection of collagen stroma peptides and could reveal peptide regulation influenced by specific cell types. Our initial work demonstrated that stromal peptide peak intensities when using MALD-IMS following IHC staining (αSMA, FAP, P4HA3 and PTEN) were comparable to serial sections of nonstained tissue. Analysis of histology-directed IMS using PTEN on breast tissues and TMAs revealed heterogeneous PTEN staining patterns and suggestive roles in stromal protein regulation. This study sets the foundation for investigations of target cell types and their unique contribution to collagen regulation within extracellular matrix niches.

Biopsy site changes in mediastinal lymph nodes (LNs) attributable to prior endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) have not been studied in a systematic manner. Twenty-four contributors from 14 institutions in 5 countries collaborated via social media (Twitter) to retrospectively review consecutive cases of resected mediastinal LNs from patients with prior EBUS-TBNA. Resected LNs were reexamined by submitting pathologists for changes attributable to EBUS-TBNA. Patients who received neoadjuvant therapy were excluded. Cases with suspected biopsy site changes underwent central review by 5 pathologists. A total of 297 mediastinal LN resection specimens from 297 patients (183 male/114 female, mean age: 65 y, range: 23 to 87) were reviewed. Biopsy site changes were most common in station 7 (10 cases) followed by 11R, 4R, and 10R, and were found in 34/297 (11.4%) cases, including displacement of tiny cartilage fragments into LN parenchyma in 26, intranodal or perinodal scars in 7, and hemosiderin in 1. Cartilage fragments ranged from 0.26 to 1.03 mm in length and 0.18 to 0.62 mm in width. The mean interval between EBUS-TBNA and LN resection was 38 days (range: 10 to 112) in cases with biopsy site changes. A control group of 40 cases without prior EBUS-TBNA, including 193 mediastinal LN stations, showed no evidence of biopsy site changes. Biopsy site changes are identified in a subset of resected mediastinal LNs previously sampled by EBUS-TBNA. The location of the abnormalities, temporal association with prior EBUS-TBNA, and the absence of such findings in cases without prior EBUS-TBNA support the contention that they are caused by EBUS-TBNA.

Purpose Lifestyle factors associated with personal behavior can alter tumor-associated biological pathways and thereby increase cancer risk, growth, and disease recurrence. Advanced glycation end products (AGEs) are reactive metabolites produced endogenously as a by-product of normal metabolism. A Western lifestyle also promotes AGE accumulation in the body which is associated with disease phenotypes through modification of the genome, protein crosslinking/dysfunction, and aberrant cell signaling. Given the links between lifestyle, AGEs, and disease, we examined the association between dietary-AGEs and breast cancer. Methods We evaluated AGE levels in bio-specimens from estrogen receptor-positive (ER+) and estrogen receptor-negative (ER−) breast cancer patients, examined their role in therapy resistance, and assessed the ability of lifestyle intervention to reduce circulating AGE levels in ER+ breast cancer survivors. Results An association between ER status and AGE levels was observed in tumor and serum samples. AGE treatment of ER+ breast cancer cells altered ERα phosphorylation and promoted resistance to tamoxifen therapy. In a proof of concept study, physical activity and dietary intervention was shown to be viable options for reducing circulating AGE levels in breast cancer survivors. Conclusions There is a potential prognostic and therapeutic role for lifestyle derived AGEs in breast cancer. Given the potential benefits of lifestyle intervention on incidence and mortality, opportunities exist for the development of community health and nutritional programs aimed at reducing AGE exposure in order to improve breast cancer prevention and treatment outcomes. Electronic supplementary material The online version of this article (10.1007/s10549-018-4992-7) contains supplementary material, which is available to authorized users.

Background This study aimed to investigate the changes in diagnosis after a second opinion for breast cancer patients from a multi-disciplinary tumor board (MTB) review at an National Cancer Institute (NCI)-designated cancer center. Methods A retrospective study analyzed patients with a breast cancer diagnosed at an outside institution who presented for a second opinion from August 2015 to March 2016 at the Medical University of South Carolina (MUSC). Radiology, pathology, and genetic testing reports from outside institutions were compared with reports generated after an MTB review and subsequent workup at MUSC. The second-opinion cases were categorized based on whether diagnostic variations were present or not. Results The review included 70 patients seeking second opinions, and 33 (47.1%) of these patients had additional radiologic images. A total of 30 additional biopsies were performed for 25 patients, with new cancers identified in 16 patients. Overall, 16 (22.8%) of the 70 of patients had additional cancers diagnosed. For 14 (20%) of the 70 patients, a second opinion led to a change in pathology interpretation. Genetic testing was performed for 11 patients (15.7%) who met the National Comprehensive Cancer Network (NCCN) guidelines for genetic testing, but none showed a mutation other than a variant of unknown significance. After a complete workup, 30 (42.8%) of the 70 patients had a change in diagnosis as a result of the MTB review. Conclusion A review by an MTB at an NCI-designated cancer center changed the diagnosis for 43% of the patients who presented for a second opinion for breast cancer. The study findings support the conclusion that referral for a second opinion is beneficial and has a diagnostic impact for many patients.

This report highlights a diagnostically challenging case of diffuse deciduoid mesothelioma occurring in the peritoneum of a 25-year-old woman, 8 months postpartum. Optimally debulked tumor consisted of sheets of polygonal cells arranged in solid, trabecular, and pseudopapillary configurations, with vesicular, occasionally grooved nuclei and small nucleoli. A barrage of immunohistochemical stains revealed an unusual staining pattern characterized by diffusely positive keratin, WT-1, and mesothelin staining, but lack of calretinin positivity. Electron microscopy demonstrated only rare thin, long, branching cellular projections. Cytogenetics revealed balanced translocations of 12p and 1q and 16p. Based on compiled ancillary studies, a diagnosis of deciduoid mesolthelioma was made and supported by consultations from experts at 3 outside facilities. Twenty-seven months after diagnosis, the patient is alive and undergoing treatment with progression of disease. This case is presented in detail, and a discussion of the diagnostic criteria and current application of those criteria is provided.

Context.— Social media sites are increasingly used for education, networking, and rapid dissemination of medical information, but their utility for facilitating research has remained largely untapped. Objective.— To describe in detail our experience using a social media platform (Twitter) for the successful initiation, coordination, and completion of an international, multi-institution pathology research study. Design.— Following a tweet describing a hitherto-unreported biopsy-related histologic finding in a mediastinal lymph node following endobronchial ultrasound–guided transbronchial needle aspiration, a tweet was posted to invite pathologists to participate in a validation study. Twitter's direct messaging feature was used to create a group to facilitate communication among participating pathologists. Contributing pathologists reviewed consecutive cases of mediastinal lymph node resection following endobronchial ultrasound–guided transbronchial needle aspiration and examined them specifically for biopsy site changes. Data spreadsheets containing deidentified data and digital photomicrographs of suspected biopsy site changes were submitted via an online file hosting service for central review by 5 pathologists from different institutions. Results.— A total of 24 pathologists from 14 institutions in 5 countries participated in the study within 143 days of study conception, and a total of 297 cases were collected and analyzed. The time interval between study conception and acceptance of the manuscript for publication was 346 days. Conclusions.— To our knowledge, this is the first time that a social media platform has been used to generate a research idea based on a tweet, recruit coinvestigators publicly, communicate with collaborating pathologists, and successfully complete a pathology study.

We previously reported that secreted frizzled-related protein-2 (SFRP2) is expressed in a variety of tumors, including sarcoma and breast carcinoma, and stimulates angiogenesis and inhibits tumor apoptosis. Therefore, we hypothesized that a humanized SFRP2 monoclonal antibody (hSFRP2 mAb) would inhibit tumor growth. The lead hSFRP2 antibody was tested against a cohort of 22 healthy donors using a time course T-cell assay to determine the relative risk of immunogenicity. To determine hSFRP2 mAb efficacy, nude mice were subcutaneously injected with SVR angiosarcoma cells and treated with hSFRP2 mAb 4 mg/kg intravenously every 3 days for 3 weeks. We then injected Hs578T triple-negative breast cells into the mammary fat pad of nude mice and treated for 40 days. Control mice received an immunoglobulin (Ig) G1 control. The SVR and Hs578T tumors were then stained using a TUNEL assay to detect apoptosis. Immunogenicity testing of hSFRP2 mAb did not induce proliferative responses using a simulation index (SI) ≥ 2.0 (p

Purpose Using a recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method, human breast cancer formalin-fixed paraffin-embedded (FFPE) tissue sections and tissue microarrays (TMA) are evaluated for N-linked glycan distribution in the tumor microenvironment. Experimental design Tissue sections representing multiple human epidermal growth factor receptor 2 (HER2) receptor-positive and triple-negative breast cancers (TNBC) in both TMA and FFPE slide format are processed for high resolution N-glycan MALDI-IMS. An additional FFPE tissue cohort of primary and metastatic breast tumors from the same donors are also evaluated. Results The cumulative N-glycan MALDI-IMS analysis of breast cancer FFPE tissues and TMAs indicate the distribution of specific glycan structural classes to stromal, necrotic, and tumor regions. A series of high-mannose, branched and fucosylated glycans are detected predominantly within tumor regions. Additionally, a series of polylactosamine glycans are detected in advanced HER2+, TNBC, and metastatic breast cancer tissues. Comparison of tumor N-glycan species detected in paired primary and metastatic tissues indicate minimal changes between the two conditions. Conclusions and clinical relevance The prevalence of tumor-associated polylactosamine glycans in primary and metastatic breast cancer tissues indicates new mechanistic insights into the development and progression of breast cancers. The presence of these glycans could be targeted for therapeutic strategies and further evaluation as potential prognostic biomarkers.

Traditionally, the radiology elective has been designed to teach medical students the fundamentals of radiologic interpretation. When questioned, many students state that they want to take a radiology elective so they can "interpret images." For the students on radiology, rotation/elective education was often passive, consisting of didactic conferences and observational shadowing of radiologists as they interpreted images. Students had only a superficial appreciation of how radiologists interacted with clinical services, multidisciplinary teams, and pathology. There was very little emphasis on imaging appropriateness or the most efficient and effective imaging for various clinical problems. With the expansion of numerous imaging modalities and the emphasis on patient-centered care, including imaging safety and dose reduction, it is important to change the focus of radiology education from interpretation to the optimal integration of imaging into clinical medicine. Radiology-pathology (rad path) electives were created at Allegheny General Hospital and the Medical University of South Carolina as a new option to provide a high-quality advanced elective for fourth-year medical students. These electives enable students to correlate radiologic images with gross and microscopic pathology specimens, thus increasing their knowledge and understanding of both. The rad path elective combines aspects of surgery, radiology, and pathology and requires students to be active learners. The implementation of this elective is an exciting work in progress that has been evolving over the past 2 and 4 years at Medical University of South Carolina and Allegheny General Hospital, respectively. We will discuss the historical basis for the elective, the advantages and challenges of having such an integrated course, and some different strategies for creating a rad path elective.

Aim Central efferent and afferent neural pathways to and from the human urinary bladder are well-characterized, but the location and arborization of these nerves as they traverse the serosa, muscularis, and urothelial layers are not clearly defined. The purpose of this study was to create a three dimensional map of the innervation of the human bladder trigone from the extrinsic perivesical adventitial nerve trunks to the urothelium. Methods A male and a female human bladder were harvested from fresh frozen cadavers and fixed in formalin. The bladder neck and trigone region were serially sectioned (5 μm) and every 20th slide was stained (S100), scanned and aligned to create 3D maps. Results Nerve penetration into the detrusor muscle occurs with the highest frequency at the bladder neck and interureteric ridge. Nerves traveling parallel to the bladder lumen do so in the adventitia, beyond the outer border of detrusor. In females, the depth of these nerve bands is uniform at 0.7–1.7 cm below the luminal surface, the outer limits of which include the anterior vaginal wall. In the male, depth is more variable owing to detrusor hypertrophy with the minimum depth of nerves approximately 0.5 cm near the interureteric ridge and over 1 cm near the bladder neck. Conclusions Myelinated neural pathways traversing in the human bladder in the region of the trigone have a discreet regional density. This 3D map of trigonal innervation may provide guidance to more precisely direct therapies for urinary incontinence or pelvic pain. Neurourol. Urodynam. © 2016 Wiley Periodicals, Inc.

Advanced glycation end products (AGEs) are reactive sugar metabolites that our research has shown can increase cancer severity. AGEs are formed endogenously via a slow, nonenzymic reaction. However, high AGE levels associate with the same socioeconomic and environmental factors that contribute to cancer health disparity. Lack of exercise and modern dietary patterns represent cancer disparity factors that lead to the increased accumulation of AGEs. Our published research indicates that AGE levels are highest in prostate cancer patients with more aggressive disease and those with African Ancestry. An accumulation of AGEs in the body drives chronic inflammation which has an impact on the progression of diseases such as Alzheimer’s, diabetes, and as we have recently shown cancer. Our unpublished data demonstrate that consumption of a diet high in AGEs increases the growth of prostate cancer in both syngeneic xenograft and spontaneous mouse models. Dietary-AGE mediated effects on prostate tumor growth were dependent upon the stromal expression of the receptor for AGE (RAGE) which led to the activation of fibroblasts. As part of a funded study (NIH/NCI: PQ3-R21CA194469 PI: Turner), we generated primary fibroblast and epithelial cell lines from Gleason 7 radical prostatectomy tissues donated by AA and EA men. Gleason 7 tumors represent a critical transition point in prostate cancer progression. Cells were molecularly characterized, confirmed as novel by STR analysis and percent ancestry confirmed. We performed two-compartment co-culture migration models with the ancestry specific primary cell lines. Compared to the corresponding experiment in EA men, we found that the presence of primary fibroblasts from AA men significantly increased the migratory potential of patient matched epithelial cells. We also examined the effects on epithelial cell migration when the ancestry specific stromal cells were cross-cultured. Compared to patient matched fibroblasts, the migratory potential of EA epithelial cells was significantly higher when cross-cultured with fibroblasts isolated from AA men. In contrast, the migratory potential of AA epithelial cells was significantly reduced when cross-cultured with EA fibroblasts. In both cases, the effects on epithelial cell migration was exacerbated when the ancestry specific fibroblasts were treated with exogenous AGE with the highest effects being observed with AA derived fibroblasts. The dietray-AGE mediated regulation of the tumor stroma in AA men with prostate cancer indicates an ancestry specific destablization of the tumor microenvironment supportive for aggressive disease. AGEs may represent a biological consequence of cancer disparity factors that can contibute to the increased prostate cancer incidence and mortality observed in men with African ancestry. Citation Format: Courtney E. Lloyd, Bradley A. Krsianits, Pamela Woods, Dion Foster, Lourdes M. Nogueira, Bria Sanders, Laura Spruill, Marvella E. Ford, Mahtabbudin Ahmed, Victoria J. Findlay, David P. Turner. Cause and effect relationships between glycation and the ancestry specific tumor stroma [abstract]. In: Proceedings of the AACR Virtual Conference: Thirteenth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2020 Oct 2-4. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(12 Suppl):Abstract nr PR10.

Lifestyle factors associated with personal behavior can alter tumor-associated biologic pathways and thereby increase cancer risk, growth and disease recurrence. Advanced glycation end products (AGEs) are reactive metabolites produced endogenously as a byproduct of normal metabolism. A Western lifestyle consisting of high-fat, high-sugar and processed foods as well as little exercise can lead to a significant increase in AGE accumulation in the body and is also associated with driving cancer disparity. Increased AGE accumulation promotes disease phenotypes through modification of the genome, protein crosslinking and dysfunction, and aberrant cell signaling. We evaluated AGE levels in biospecimens from ER+ and ER- breast cancer patients, examined their role in therapy resistance, and assessed the ability of a lifestyle intervention to reduce circulating AGE levels in ER+ breast cancer survivors. A correlation between ER status and AGE levels was observed in tumor and serum samples. AGE treatment of ER+ breast cancer cells impacted pathways associated with ER regulation. We observed a significant increase in phosphorylation of ERalpha following AGE treatment when compared to untreated control with no change in total ERalpha levels. We also observed a significant increase in both AKT and ERK phosphorylation in ER+ cell lines in response to AGE treatment in a time-dependent manner. Inhibition of AKT with Ly294002 and inhibition of ERK with the MEK inhibitor U0126 significantly reduced ERalpha phosphorylation in the presence of AGE. Significantly, ER+ cells treated with AGEs no longer responded to hormonal therapy with tamoxifen. In a proof-of-concept study we examined the ability of a defined exercise and dietary intervention (i.e., cardiac rehabilitation) to reduce circulatory AGE levels in ER+ breast cancer survivors. A significant increase in average very active minutes and average calories burned was observed as a result of the intervention. This was accompanied by a significant reduction in dietary-AGE intake and also showed significant reductions in circulating AGE levels when fasting serum samples were analyzed by ELISA. An analysis of IL6 and CRP levels by ELISA in the same AGE assessed samples revealed no significant differences at any time point. There is a potential prognostic and therapeutic role for lifestyle-derived AGEs in cancer disparity. Given the potential benefits of lifestyle intervention on cancer incidence and mortality, opportunities exist for the development of community health and nutritional programs aimed at reducing AGE exposure in order to improve cancer prevention and treatment outcomes. Lifestyle interventions that lower AGE levels may then be utilized to reduce breast cancer incidence and improve prognosis in cancer disparity populations. Citation Format: Katherine R. Walter, Ford E. Ford, Mathew J. Gregoski, Rita M. Kramer, Kendrea D. Knight, Laura Spruill, Lourdes M. Nogueira, Bradley A. Krisanits, Marian H. Taylor, Amanda C. La Rue, Michael B. Lilly, Stefan Ambs, King Chan, Tonya F. Turner, Heidi Varner, Shweta Singh, Jaime Uribarri, Elizabeth Garrett-Mayer, Kent E. Armeson, Ebony J. Hilton, Mark Clair, Victoria J. Findlay, Lindsay L. Peterson, Gayenell Magwood, David P. Turner. Lifestyle-associated advanced glycation end products are elevated in ER+ positive breast cancer patients, alter response to therapy, and can be targeted by lifestyle intervention [abstract]. In: Proceedings of the Eleventh AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2018 Nov 2-5; New Orleans, LA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl):Abstract nr C024.

Changes in cell surface protein glycosylation are common alterations that occur with tumor progression and reflect the use of many cancer biomarkers like PSA and CA19-9. However, identification of specific glycans associated with tumor regions is still poorly defined. Our group has developed a two-dimensional glycan tissue imaging mass spectrometry approach that can be used with any clinical formalin-fixed, paraffin-embedded tumor tissue used in pathology. Based on the analysis of over one thousand prostate cancer FFPE tissue blocks and tissue microarray samples across the spectrum of disease, it was found that the presence of multi-fucosylated branched N-glycans is associated with advanced tumors with neuroendocrine and metastatic features. These glycans are not present in indolent and lower-grade adenocarcinomas. The hypothesis currently being tested is to determine whether detection of higher numbers of these fucosylated structures predicts a worse prognosis for progressive metastatic disease. Race could be a factor for increased presentation at the time of diagnosis with more advanced disease. The goal of the present study was to assess whether more advanced tumors containing multi-fucosylated N-glycans are detected more frequently in a racial cohort of 307 samples. A series of 14 prostate tumor tissue microarrays (TMA) representing African-American (n=105), Hispanic (n=101) and Caucasian (n=101) subjects was evaluated with this method. The 307 individual tumor samples represented the spectrum of tumor stage, Gleason grade, and status of disease recurrence. For select cases, the original source tumor block tissues were analyzed to confirm the TMA core results. Each TMA slide was processed for antigen retrieval and peptide N-glycosidase F digestions to release N-glycans. Samples were analyzed by MALDI-FTICR mass spectrometry, and data were visualized and analyzed by SCiLS Lab software. Tumor tissues having increased levels of fucosylation were detected in each cohort, but with an overall increase in numbers detected in the African-American cohort. Multitiered analyses are ongoing to identify glycan signatures associated with age, grade, stage, race, and recurrence. In summary, the presence and detection of a distinct panel of multi-fucosylated tissue N-glycans associated with the most lethal forms of prostate cancer can be detected in low-grade tumor samples at the time of diagnosis. This could be developed into a tissue-based prognostic biomarker panel to be applied at the time of initial diagnosis, and impact earlier treatment decisions. Citation Format: Richard R. Drake, Fred David, Cameron Miller, Melanie Jefferson, Laura Spruill, Michael Liss, Brandi Weaver, Peggi M. Angel, Robin Leach, Chanita Hughes-Halbert. Distinguishing lethal from indolent prostate cancer using N-glycan imaging mass spectrometry in a racial tissue microarray cohort [abstract]. In: Proceedings of the Eleventh AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2018 Nov 2-5; New Orleans, LA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl):Abstract nr C032.

Advanced glycation end-products (AGEs), are reactive metabolites produced endogenously as a consequence of glucose uptake during glycolysis. AGEs accumulate in tissues and organs as we grow older to promote multiple chronic disease phenotypes. AGE pathogenic effects are mediated through modification of protein function, genetic fidelity, stress responses and cellular signaling pathways. Critically, cancer disparity factors such as a sedentary lifestyle, obesity and an unhealthy diet are external influences that also contribute to the AGE accumulation pool in the body. This research group examined circulating and tumor AGE levels in clinical specimens of prostate cancer and identified a race specific, tumor-dependent pattern of accumulation. AGE levels were highest in aggressive tumors, especially those from men with African ancestry. Increased AGE levels correlated with an upregulation in the receptor for advanced glycation end products (RAGE) and activated NFkB. In a syngeneic subcutaneous prostate cancer mouse model, chronic consumption of AGE resulted in a 3-fold increase in tumor growth. Strikingly, dietary-AGE mediated increases in tumor growth were accompanied by a cytoplasmic accumulation of AR, elevation in MYC, RAGE, and AGE as well as increased cell proliferation. Cytoplasmic accumulation of AR is elevated in CRPC tissue and is an independent predictor of biochemical recurrence. Our data show that treatment of prostate cancer cells with AGEs induces a neuroendocrine differentiated phenotype by promoting a more mesenchymal morphology and increasing the expression of associated marker genes including ENO2, MYC and SYP and the prostate cancer stem cell marker CD44 as well as the downregulation of AR. The aberrant activation and recruitment of immune cells is a major pathogenic consequence of AGE accumulation, and a series of studies have highlighted the tumor-associated immune response as a critical pathway contributing to cancer disparity. Using patient-derived primary tumor cells, the investigators found that AGEs released into the extracellular matrix may recapitulate the tumor-associated immune response observed in ancestry-specific prostate tumor tissues. Further preliminary studies indicate that AGE treatment of prostate cancer cells can alter how cancer cells metabolize glucose to promote an aggressive phenotype. The investigators' studies support the concept that AGE metabolites represent a biologic consequence of the socioeconomic and environmental factors that promote cancer health disparity. As our understanding of tumor biology advances, it is becoming increasingly clear that these inter-related lifestyle factors have distinct molecular consequences on the biologic makeup of tumors, altering cell signaling events and gene expression profiles to contribute to cancer disparity outcomes such as its earlier development or its progression to more aggressive disease. Citation Format: Pamela Woods, Bradley A. Krisanits, Dion Foster, Lourdes M. Nogueira, Laura Spruill, Marvella E. Ford, Michael B. Lilly, Victoria J. Findlay, David P. Turner. Advanced glycation end products promote prostate tumor growth and are a potential biologic consequence of lifestyle factors contributing to cancer disparity [abstract]. In: Proceedings of the Eleventh AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2018 Nov 2-5; New Orleans, LA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl):Abstract nr B047.

Advanced glycation end-products (AGEs), are reactive metabolites produced endogenously as a consequence of glucose metabolism. AGEs accumulate in tissues and organs as we grow older to promote multiple chronic disease phenotypes. AGE pathogenic effects are mediated through modification of protein function, genetic fidelity, stress responses and cellular signaling pathways. Critically, cancer disparity factors such as a sedentary lifestyle, obesity and an unhealthy diet are external influences that have been shown to contribute to the accumulation of AGEs. This research group examined circulating and tumor AGE levels in clinical specimens of prostate cancer and identified a race specific, tumor-dependent pattern of accumulation. AGE levels were highest in aggressive tumors, especially those from men with African ancestry. As our understanding of tumor biology advances, it is becoming increasingly clear that these lifestyle factors have distinct molecular consequences on the biologic make up of tumors, altering cell signaling events and gene expression profiles to contribute to cancer disparity outcomes such as earlier development or its progression to more aggressive disease. Increased AGE levels correlated with an up-regulation in the receptor for advanced glycation end products (RAGE) and activated NFkB. In a syngeneic sub-cutaneous prostate cancer mouse model, chronic consumption of AGE resulted in a 3-fold increase in tumor growth. Dietary-AGE mediated increases in tumor growth were accompanied by a cytoplasmic accumulation of AR, elevation in MYC, RAGE, and AGE as well as increased cell proliferation. Given the links between lifestyle and AGEs we examined the effects of regular physical activity on AGE induced tumor growth in our syngeneic sub-cutaneous dietary-AGE prostate cancer model. Mice exposed to physical activity for 1 hour, 5 days per week showed a significant decrease in AGE induced tumor growth. We also examined the effects of dietary-AGEs on tumor progression using the FVB-Tg(C3-1-TAg)cJeg/JegJ (C3-Tag) transgenic spontaneous prostate cancer mouse model. This model progresses to low grade prostate intraepithelial neoplasia (PIN) at 24 weeks. However, chronic consumption of AGE resulted in increased progression towards moderate to high grade PIN at this same time point. When regular physical activity was introduced, we observed delayed progression of PIN in both dietary groups, but most significantly in the high AGE fed mice. These studies support the concept that AGEs represent a biological consequence of the socioeconomic and environmental factors that promote cancer disparity, which may be at least in part reversed via physical activity. This may have the greatest impact for African American patients who tend to have poorer survival, and where a lack of physical activity, poor diet, and high obesity rates are more prevalent. Citation Format: Bradley A Krisanits, Pamela M Woods, Dion Foster, Lourdes M Nogueira, Laura Spruill, Marvella E Ford, Victoria J Findlay, David P Turner. Regular physical activity can prevent the oncogenic effects of lifestyle-associated advanced glycation end products [abstract]. In: Proceedings of the Twelfth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2019 Sep 20-23; San Francisco, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl_2):Abstract nr C031.

OBJECTIVES: Lifestyle factors such as a sedentary lifestyle, obesity and a diet consisting of foods high in protein, fat, and highly processed foods contribute to the accumulation of advanced glycation end products (AGEs). AGE pathogenic effects are mediated through modification of protein function, genetic fidelity, stress responses, and cellular signaling pathways which can promote the development of a variety of chronic diseases. However, a gap in the literature exists on the role of dietary AGE in cancer progression and the potential impact of physical activity (PA) to negate such effects. Given the links between lifestyle and AGEs, we propose that a diet high in AGEs can accelerate prostate cancer progression and can be reversed by PA. METHODS: Xenograft and spontaneous prostate cancer mouse models were utilized to assess the effects of dietary AGE on prostate tumor growth. Mice were exposed to PA on an Animal Treadmill for 1 hr, 5 days a week to assess the impact on dietary AGE mediated tumor progression. Tumor and prostate tissue were collected for histology, and Western Blot analysis. PBMCs were isolated from endpoint blood draws for flow cytometric analysis. RESULTS: We found that chronic consumption of AGEs leads to a 3-fold increase in tumor growth in xenograft mice, accompanied by decreased expression of AR, increased expression of MYC, RAGE, and AGE as well as cell proliferation. In the spontaneous prostate cancer model chronic consumption of AGE resulted in increased progression towards high grade PIN and carcinoma when compared to l mice. IHC showed that the high AGE fed mice had increased recruitment of macrophages to PIN lesions. Mice in the PA arm showed a decrease in tumor growth (xenograft) and delayed progression (spontaneous). This was observed in all diet groups, but was most significant in the dietary AGE mice. CONCLUSIONS: Our studies support the concept that AGEs represent a biological consequence of lifestyle factors that promote cancer progression, and that PA may alleviate these effects. This may have the greatest impact in African American populations who have worse outcomes in prostate cancer, and where a lack of PA, poor diet, and high obesity rates are more prevalent. FUNDING SOURCES: Bradley Krisanits was supported by Hollings Cancer Center Pre-Doctoral Fellowship (MUSC), Graduate Assistance in Areas of National Need (US Dept. of Ed.), and U54 CA21096.

Tissue necrosis is a form of cell death common in advanced and aggressive solid tumors, and is associated with areas of intratumoral chronic ischemia. The histopathology of necrotic regions appear as a scaffold of cellular membrane remnants, reflective of the hypoxia and cell degradation events associated with this cellular death pathway. Changes in the glycosylation of cell surface proteins is another common feature of cancer progression. Using a recently developed mass spectrometry imaging approach to evaluate N-linked glycan distributions in human formalin-fixed clinical cancer tissues, differences in the glycan structures of regions of tumor, stroma and necrosis were evaluated. While the structural glycan classes detected in the tumor and stromal regions are typically classified as high mannose or branched glycans, the glycans found in necrotic regions displayed limited branching, contained sialic acid modifications and lack fucose modifications. While this phenomenon was initially classified in breast cancer tissues, it has been also seen in cervical, thyroid and liver cancer samples. These changes in glycosylation within the necrotic regions could provide further mechanistic insight to necrotic changes in cancer tissue and provide new research directions for identifying prognostic markers of necrosis.

Objectives To develop a urodynamic model incorporating external urethral sphincter (EUS) electromyography (EMG) in awake rats. Materials and Methods Bladder catheters and EUS EMG electrodes were implanted in female Sprague Dawley rats. Assessments were performed in awake, lightly restrained rats on postoperative day 12–14. Measurements were repeated in the same rat on day 16 under urethane anaesthesia. Urodynamics and EUS EMG were performed simultaneously. In addition, serum creatinine and bladder histology was assessed. Results No significant differences in urodynamic parameters were found between bladder catheter only vs bladder catheter and EUS EMG electrode groups. Urethane anaesthesia evoked prominent changes in both urodynamic parameters and EUS EMG. Serum creatinine was within the normal limits in all rats. Bladder weight and bladder wall thickness were significantly increased in both the bladder catheter only and the bladder catheter and EUS EMG group compared with controls. Conclusions Our novel urodynamic model allows repetitive measurements of both bladder and EUS function at different time points in the same rat under fully awake conditions and opens promising avenues to investigate lower urinary tract dysfunction in a translational approach.

NSD3 has been implicated as a candidate driver oncogene from the 8p11-p12 locus, and we have previously published evidence for its amplification and overexpression in human breast cancer. This aim of this study was to further characterize the transforming function of NSD3 in vivo. We generated a transgenic mouse model in which NSD3 gene expression was driven by the MMTV promoter and expressed in mammary epithelium of FVB mice. Mammary glands were fixed and whole mounts were stained with carmine to visualize gland structure. Mammary tumors were formalin-fixed, and paraffin embedded (FFPE) tumors were stained with hematoxylin and eosin. Pups born to transgenic females were significantly underdeveloped compared to pups born to WT females due to a lactation defect in transgenic female mice. Whole mount analysis of the mammary glands of transgenic female mice revealed a profound defect in functional differentiation of mammary gland alveoli that resulted in the lactation defect. We followed parous and virgin NSD3 transgenic and control mice to 50 weeks of age and observed that several NSD3 parous females developed mammary tumors. Whole mount analysis of the mammary glands of tumor-bearing mice revealed numerous areas of mammary hyperplasia and ductal dysplasia. Histological analysis showed that mammary tumors were high-grade ductal carcinomas, and lesions present in other mammary glands exhibited features of alveolar hyperplasia, ductal dysplasia, and carcinoma in situ. Our results are consistent with our previous studies and demonstrate that NSD3 is a transforming breast cancer oncogene.

African American (AA) females with endometrial carcinoma have a significantly worse prognosis with regard to disease-free survival and overall survival than their European American (EA) counterparts and this finding is true across all stages and grades. The presence of tumor-infiltrating lymphocytes (TILs) has been demonstrated to be of prognostic significance in a variety of malignancies, including endometrial cancers. This study aims to determine whether clinically significant differences in levels of CD8+ cytotoxic T lymphocytes, FoxP3+ regulatory T lymphocytes, and CD45RO+ memory T lymphocytes exist between races and to document the clinical impact of TILs. One hundred ten patients with endometrial adenocarcinoma, treated with hysterectomy from 2003 to 2011 were studied. Patients were selected to provide equal representation across type and grade for both EAs and AAs. Immunohistochemical stains were used to highlight CD8-positive, FoxP3-positive, and CD45RO-positive TILs at the endometrial-myometrial interface on slides from paraffin-embedded tissue. Patients with "high" or "low" levels of TILs were compared with respect to the race, tumor type, and survival. High levels of CD45RO+ TILs were associated with improved overall survival in EA women (hazard ratio, 0.32; 95% confidence interval, 0.11-0.92; P=0.034). Comparatively, AA women with high levels of CD45RO+ TILs received no survival benefit (hazard ratio, 0.96; 95% confidence interval, 0.35-2.64; P=0.94). High levels of CD8-positive or FoxP3-positive TILs, alone, had no impact on survival. EA patients with TILs containing high levels of CD45RO cells but low levels of CD8+ cells lost the survival benefit; however, limited numbers preclude significant conclusions from this observation. Neither tumor type nor race were predictive of the levels of TILs of any type. Further study with a larger sample size is required to determine the impact of TIL subtype combinations on survival.

Socioeconomic and environmental influences are established factors promoting cancer disparity, but the contribution of biologic factors is not clear. We report a mechanistic link between carbohydrate-derived metabolites and cancer that may provide a biologic consequence of established factors of cancer disparity. Glycation is the nonenzymatic glycosylation of carbohydrates to macromolecules, which produces reactive metabolites called advanced glycation end products (AGE). A sedentary lifestyle and poor diet all promote disease and the AGE accumulation pool in our bodies and also increase cancer risk. We examined AGE metabolites in clinical specimens of African American and European American patients with prostate cancer and found a higher AGE concentration in these specimens among African American patients when compared with European American patients. Elevated AGE levels corresponded with expression of the receptor for AGE (RAGE or AGER). We show that AGE-mediated increases in cancer-associated processes are dependent upon RAGE. Aberrant AGE accumulation may represent a metabolic susceptibility difference that contributes to cancer disparity. Cancer Epidemiol Biomarkers Prev; 23(10); 2186–91. ©2014 AACR.

Pilomatrixoma or calcifying epithelioma of Malherbe is a benign skin tumor arising from the hair follicle; breast occurrence is considered a rarity. Clinically presenting as a palpable abnormality and with both benign and malignant mammographic and sonographic features, it can be easily misdiagnosed as a breast neoplasm. We report a very rare case of pilomatrixoma of the male breast in a 36-year-old male presenting with a firm, superficial nodule in the upper outer quadrant. Though the sonographic trifecta of imaging features (shape- margins-orientation/oval, circumscribed mass, parallel to the skin) is consistent with a benign lesion, a histologic diagnosis was warranted based on its most suspicious feature of internal pleomorphic calcifications. Pathologic diagnosis revealed the uncommon benign entity of pilomatrixoma in the male breast. Our patient was recommended for surgical excision based on current literature recommendations for management in most reports of pilomatrixoma. One alternative recommendation presented in a single report of pilomatrixoma in the breast supported follow-up imaging based on benign imaging characteristics.

Background Liposarcomas are rare in the head and neck. We analyzed a large series of head and neck liposarcomas to determine features unique to the head and neck. Methods Three hundred eighteen liposarcomas of the head and neck were contrasted with 9485 liposarcomas of other regions using the Surveillance, Epidemiology, and End Results (SEER) database. Results Head and neck liposarcomas were most commonly subcutaneous (81.%), low grade (70.1%; p < .001), and early stage (p < .001). They were more likely to be treated with surgery alone, whereas conventional liposarcomas were more likely to receive adjuvant radiation (p < .001). Treatment that included surgery had better survival than radiation therapy alone (p = .008). Overall, liposarcomas of the head and neck had significantly higher disease-specific survival (DSS) and overall survival (OS) than conventional liposarcomas (p < .001). Conclusion Liposarcomas of the head and neck are usually early stage, low grade, and with fewer nodal metastases than conventional liposarcomas. DSS and OS were significantly greater for liposarcomas of the head and neck. © 2013 Wiley Periodicals, Inc. Head Neck 36: 393–400, 2014

Aman in his 50s presented with a 2-month history of a rapidly enlargingmass on the left side of the neck. He reported a recent 5-kg weight loss and had no history of tobacco or alcohol use. Clinical examination revealed a fixed lower neckmass extending through levels 3, 4, 5b, and6,measuring 7 × 8 cm (Figure, A). Flexible laryngoscopy revealed reduced movementof the leftvocalcordandasymmetrywithsurrounding edema. Computed tomographic imaging revealed a well-vascularizedmass of the paratracheal region invading surrounding the surrounding structures (Figure,B).Twofine-needleaspirationsampleswereobtained, but results from pathological analysis were inconclusive. The tumor was removed in a piecemeal fashion because it surrounded the trachea and esophagus and invaded into the strap muscles, internal jugular vein, the first tracheal ring, and the recurrent laryngeal nerve. At the end of the procedure all margins were clear. On pathologic examination, the tumorwas semifirmwith a necrotic central portion. The hematoxylin-eosin–stained specimen revealed many mitotic figures along with large, multinucleated giant cells. Lymphovascular andperineural invasionwas present (Figure, C). Immunohistochemical staining of themainspecimenwascytokeratinAE1/AE3negativeandthyroid transcription factor (TTF-1) negative (Figure, D). Of 12 lymph nodes, none contained carcinoma. Quiz at jamaotolaryngology.com Photograph of neck mass A Computed tomographic scan of neck B

Advanced disease accounts for the majority of prostate cancer-related deaths and androgen deprivation therapy (ADT) is the standard of care for these patients. Many patients undergoing ADT become resistant to its effects and progress to castrate-resistant prostate cancer (CRPC). Current therapies for CRPC patients are inadequate, with progression-free survival rates as low as 2 months. The molecular events that promote CRPC are poorly understood. ETS (v-ets erythroblastosis virus E26 oncogene) transcription factors are regulators of carcinogenesis. Protein levels of the archetypical ETS factor, ETS1, are increased in clinical and latent prostate cancer relative to benign prostatic hyperplasia and normal prostate to promote multiple cancer-associated processes, such as energy metabolism, matrix degradation, survival, angiogenesis, migration and invasion. Our studies have found that ETS1 expression is highest in high-grade prostate cancer (Gleason 7 and above). Increased ETS1 expression and transcriptional activity promotes an aggressive and castrate-resistant phenotype in immortalized prostate cancer cells. Elevated AKT (v-akt murine thymoma viral oncogene homolog) activity was demonstrated to increase ETS1 protein levels specifically in castrate-resistant cells and exogenous ETS1 expression was sufficient to rescue invasive potential decreased by inhibition of AKT activity. Significantly, targeted androgen receptor activity altered ETS1 expression, which in turn altered the castrate-resistant phenotype. These data suggest a role for oncogenic ETS1 transcriptional activity in promoting aggressive prostate cancer and the castrate-resistant phenotype.

Uterine leiomyomas are the most common pelvic tumor, and a frequent indication of the need for gynecologic surgery. Although usually asymptomatic, life-threatening cases can occur. We present a case of critical hypercalcemia associated with a leiomyoma during pregnancy with the intention of highlighting the endocrinology of leiomyomas, features shared with malignant neoplasms, and the potential for effects on obstetric outcomes.A 32-year-old gravid woman with a large leiomyoma presented at 33 5/7 weeks of gestation with critical hypercalcemia requiring intensive care. Postpartum myomectomy cured her hypercalcemia, which was driven by parathyroid hormone-related protein (PTHrP) produced by the tumor.Obstetricians should be aware of the existence of humoral hypercalcemia related to leiomyomas and the potential effects on pregnancy.

It has been hypothesized that translational efficiency is determined by the amount of secondary structure in the 5′-untranslated region (5′-UTR) of mRNA. Here, we examined whether specific 5′-UTRs with excessive secondary structure selectively regulate translational efficiency in adult cardiocytes. Recombinant adenoviruses were generated to express reporter mRNAs consisting of the 5′-UTR derived from c-jun or ornithine decarboxylase (ODC) fused to β-galactosidase (βGal) coding sequence. Each adenovirus expressed GFP mRNA as a control for 5′-UTRs with minimal secondary structure. Subsequently, cardiocytes were electrically stimulated to contract at 1 Hz to accelerate protein synthesis as compared to quiescent controls. Translational efficiency was calculated by measuring protein expression as a function of mRNA levels. Translational efficiency of c-jun/βGal mRNA increased significantly by 3.7-fold in contracting vs. quiescent cardiocytes, but ODC/βGal mRNA was unchanged. Contraction increased c-jun/βGal mRNA levels in polyribosomes by 2.3-fold, which indicates that translational efficiency was enhanced by mobilization. A short, unstructured 5′-UTR was sufficient for efficient translation of βGal mRNA in quiescent and contracting cardiocytes. GFP mRNA produced similar results. These studies demonstrate that the 5′-UTR functions as a determinant of translational efficiency of specific mRNAs, such as c-jun, that regulate growth of the adult cardiocyte.—Spruill, L. S., McDermott, P. J. Role of the 5′-untranslated region in regulating translational efficiency of specific mRNAs in adult cardiocytes.

Breast pathology is filled with pitfalls, including underdiagnosis of bland-appearing lesions, both invasive and non-invasive, misdiagnosis of malignant lesions as belonging to the wrong subgroup, for example, calling LCIS as DCIS or missing the metaplastic component of an invasive lesion, and overdiagnosis of benign lesions as malignancy. While each is a sin of varying severity, the overdiagnosis of benign lesions can be the most scarring, especially in this age where Angelina Jolie׳s prophylactic mastectomy is the headline news and patients are pushing for aggressive preventive treatment. In this review, we will consider some of the more common benign lesions and the malignant counterpart that they mimic, with the goal of identifying characteristic features that will lead to the correct diagnosis. Much of the discussions will center around the assessment of core biopsies, as smaller tissue volume is most often contributory to overcalling benign lesions.

A 71-year-old white woman was admitted for the evaluation of progressive dyspnea. Her medical history was remarkable for 3 episodes of pneumonia and asthma with peripheral eosinophilia. As a part of her diagnostic evaluation, a contrast-enhanced computed tomographic scan of the chest was performed, which excluded pulmonary embolism and interstitial lung disease. Transthoracic echocardiogram revealed right-sided chamber enlargement, with a mildly hypokinetic right ventricle and an estimated right ventricle systolic peak pressure of 58 mm Hg. Moreover, an area of increased echogenicity was noted at the apex of the left ventricle (LV) consistent with possible LV apical mass (Movie I in the Data Supplement). Cardiac magnetic resonance imaging (MRI) (Siemens Magnetom Avanto 1.5T VB15; Siemens Healthcare, Erlangen, Germany) was obtained to characterize the mass further. Cine MR images (Movie II in the Data Supplement) confirmed the presence of an apical LV mass with obliteration of the midventricular and apical segments and associated regional dysfunction. After the administration of 0.1 mmol/kg gadopentetate dimeglumin (Magnevist, Bayer, Germany), early postcontrast images (Figure 1A) demonstrated the avascular nature of the mass consistent with thrombus. Delayed images (Figure 1B and 1C) revealed diffuse subendocardial hyperenhancement, most apparent in the …

Nkx2-5 gene mutations cause cardiac abnormalities, including deficits of function in the atrioventricular conduction system (AVCS). In the chick, Nkx2-5 is elevated in Purkinje fiber AVCS cells relative to working cardiomyocytes. Here, we show that Nkx2-5 expression rises to a peak as Purkinje fibers progressively differentiate. To disrupt this pattern, we overexpressed Nkx2-5 from embryonic day 10, as Purkinje fibers are recruited within developing chick hearts. Overexpression of Nkx2-5 caused inhibition of slow tonic myosin heavy chain protein (sMHC), a late Purkinje fiber marker but did not affect Cx40 levels. Working cardiomyocytes overexpressing Nkx2-5 in these hearts ectopically up-regulated Cx40 but not sMHC. Isolated embryonic cardiomyocytes overexpressing Nkx2-5 also displayed increased Cx40 and suppressed sMHC. By contrast, overexpression of a human NKX2-5 mutant did not effect these markers in vivo or in vitro, suggesting one possible mechanism for clinical phenotypes. We conclude that a prerequisite for normal Purkinje fiber maturation is precise regulation of Nkx2-5 levels.

Amplification within the 8p11-12 locus occurs in approximately 15% of breast cancer (BC) and is associated with poor survival and distant recurrence. Overexpression of many of the genes within the 8p11-12 region can confer a pathological gain of function to breast cells. We and others have previously demonstrated that NSD3 (WHSC1L1), a lysine methyltransferase, can act as a potent transforming oncogene. Predictions by the GISTIC algorithm of the Human Cancer Genome Atlas data suggest that NSD3 is a driving gene within 8p11-12 in multiple tumor types, including BC. We previously showed that NSD3 can regulate the expression of influential genes, such as estrogen receptor alpha (ESR1/ERα). To further explore the role of NSD3 in promoting BC, we generated FVB/N transgenic mice with targeted overexpression of NSD3 in the mammary gland and compared them to matched non-transgenic wild-type (WT) females. We observed that pups nursed by transgenic females were underdeveloped, regardless of genotype. Underdeveloped pups displayed delayed hair growth and eye opening, and were half the weight of pups nursed by WT females. To investigate this phenotype, we characterized thoracic and inguinal mammary glands from virgin, mid-pregnancy, lactating, and post-lactating mice by whole mount and histological analysis. Mammary glands from virgin transgenic females displayed increased branching and terminal bud formation. Alveolar buds from mid-pregnant transgenic glands were more numerous and densely packed. Glands from lactating transgenic females showed large areas in which the alveoli failed to undergo functional differentiation, resulting in a lactation defect. Mammary glands taken post-lactation exhibited areas of ductal and alveolar hyperplasia. At 40 weeks of age, multiple transgenic mice had palpable tumors which were excised at 1 cm3 size. Whole mount and histological analysis of glands from tumor bearing mice commonly demonstrated hyperplasia, dysplasia, and carcinoma in situ, contrary to age-matched WT glands. The areas of ductal dysplasia and carcinoma in situ closely resembled several patterns commonly observed in human breast cancer, including micropapillary, cribriform, and high grade areas of ductal carcinoma in situ. Mammary tumors were analogous to infiltrating ductal carcinomas, many of which were high grade tumors. In summary, overexpression of NSD3 in the mouse mammary gland elicited drastic deformation, inhibited functional differentiation, and caused tumor formation. Continued characterization of this mouse model, the oncogenic role of NSD3, and other mechanistic studies are essential to improve patient outcome in 8p11-12 altered cancers. Citation Format: Alex C. Rutkovsky, Brittany Turner-Ivey, Ericka L. Smith, Laura S. Spruill, Jamie N. Mills, Stephen P. Ethier. Development of mammary hyperplasia, dysplasia, and invasive ductal carcinoma in transgenic mice expressing the 8p11 amplicon oncogene NSD3 (WHSC1L1) [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 1835. doi:10.1158/1538-7445.AM2017-1835

Poor diet, low income, obesity, and a lack of exercise are established lifestyle factors that are known to increase cancer burden and are often more prevalent in African American communities. As our understanding of tumor biology advances, it is becoming increasingly clear that these inter-related lifestyle factors have distinct molecular consequences on the biologic make up of tumors, altering cell signaling events and gene expression profiles to contribute to cancer disparity outcomes such as its earlier development or its progression to more aggressive disease. Advanced glycation end-products (AGEs), are reactive metabolites produced endogenously as a consequence of glucose uptake during glycolysis. AGEs accumulate in tissues and organs as we grow older to promote multiple chronic disease phenotypes. AGE pathogenic effects are mediated through modification of protein function, genetic fidelity, stress responses and cellular signaling pathways. Critically, cancer disparity factors such as a sedentary lifestyle, obesity and an unhealthy diet are external influences that also contribute to the AGE accumulation pool in the body. The investigators studies support the concept that AGE metabolites represent a biological consequence of the socioeconomic and environmental factors that promote cancer health disparity. This research group examined circulating and tumor AGE levels in clinical specimens of prostate cancer and identified a race specific, tumor-dependent pattern of accumulation. The aberrant activation and recruitment of immune cells is a major pathogenic consequence of AGE accumulation and a series of studies have highlighted the tumor associated immune response as a critical pathway contributing to cancer disparity. Using patient derived primary tumor cells, the investigators found that AGEs released into the extracellular matrix can recapitulate the tumor associated immune response observed in race specific prostate tumor tissues. Activated immune cells show a similar metabolic profile as a glycolytic tumor cell with a shift towards increased glucose metabolism and aerobic glycolysis (i.e. the Warburg effect). Evidence suggests that abnormal glucose uptake may occur earlier in African American cancer patients with aggressive disease. Further preliminary studies indicate that AGE treatment of prostate cancer cells can alter how cancer cells metabolize glucose to promote an aggressive phenotype. Based on associations between active metabolism, lifestyle, and race, increases in AGE accumulation may represent a novel biologic mechanism contributing to cancer disparity and may represent a new paradigm to explaining the increased cancer incidence and mortality figures observed within health disparity populations. Given the potential benefits of lifestyle changes and the potential biological role of AGEs in promoting cancer, opportunities exist for collaborations impacting basic, translational, epidemiological and cancer prevention initiatives. Citation Format: Danzell Smith, Dion Foster, Laura Spruill, Lourdes Nogueira, Bradley Krisanits, Scott Cramer, Marvella Ford, Stephen Savage, Thomas Keane, Victoria Findlay, David Turner. The molecular implications of lifestyle associated metabolites (AGEs) to prostate cancer disparity. [abstract]. In: Proceedings of the Ninth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2016 Sep 25-28; Fort Lauderdale, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2017;26(2 Suppl):Abstract nr PR11.

Bladder inflammation (cystitis) underlies numerous bladder pathologies and is elicited by a plethora of agents such as urinary tract infections, bladder outlet obstruction, chemotherapies, and catheters. Pattern recognition receptors [Toll-like receptors (TLRs) and Nod-like receptors (NLRs)] that recognize pathogen- and/or damage-associated molecular patterns (PAMPs and/or DAMPs, respectively) are key components of the innate immune system that coordinates the production (TLRs) and maturation (NLRs) of proinflammatory IL-1β. Despite multiple studies of TLRs in the bladder, none have investigated NLRs beyond one small survey. We now demonstrate that NLRP3 and NLRC4, and their binding partners apoptosis-associated speck-like protein containing a COOH-terminal caspase recruitment domain (ASC) and NLR family apoptosis inhibitory protein (NAIP), are expressed in the bladder and localized predominantly to the urothelia. Activated NLRs form inflammasomes that activate caspase-1. Placement of a NLRP3- or NLRC4-activating PAMP or NLRP3-activating DAMPs into the lumen of the bladder stimulated caspase-1 activity. To investigate inflammasomes in vivo, we induced cystitis with cyclophosphamide (CP, 150 mg/kg ip) in the presence or absence of the inflammasome inhibitor glyburide. Glyburide completely blocked CP-induced activation of caspase-1 and the production of IL-1β at 4 h. At 24 h, glyburide reduced two markers of inflammation by 30–50% and reversed much of the inflammatory morphology. Furthermore, glyburide reversed changes in bladder physiology (cystometry) induced by CP. In conclusion, NLRs/inflammasomes are present in the bladder urothelia and respond to DAMPs and PAMPs, whereas NLRP3 inhibition blocks bladder dysfunction in the CP model. The coordinated response of NLRs and TLRs in the urothelia represents a first-line innate defense that may provide an important target for pharmacological intervention.

Poor diet, low income, obesity and a lack of exercise are established lifestyle factors that are known to increase cancer burden and are often more prevalent in African American communities. As our understanding of tumor biology advances it is becoming increasingly clear that these inter-related lifestyle factors have distinct molecular consequences on the biological make-up of tumors, altering cell signaling events and gene expression profiles to contribute to cancer disparity outcomes such as its earlier development or its progression to more aggressive disease. Sparse information exists about the genetic and biological factors that contribute to differential cancer survival and mortality rates observed in minority populations. A greater understanding of the interplay between risk factors and the molecular mechanisms associated with cancer disparity will significantly impact minority health. Advanced glycation end products (AGEs) are reactive metabolites produced as a by-product of sugar metabolism. Failure to remove these highly reactive metabolites can lead to protein damage, aberrant cell signaling, increased stress responses, and decreased genetic fidelity. Critically, AGE accumulation is also directly affected by our lifestyle choices such as poor diet, low income, obesity and a lack of exercise. We recently reported a potential mechanistic link between AGEs and prostate cancer which may provide a molecular consequence of our lifestyle choices that can directly impact tumor biology and contribute to cancer disparity. We examined circulating and intra-tumoral AGE metabolite levels in clinical specimens and identified a race specific, tumor dependent pattern of accumulation in prostate cancer serum and tumor. Further mechanistic studies in immortalized prostate cancer cell lines show that AGE treatment increases the expression of the receptor for AGEs (RAGE) to activate cancer-associated signaling cascades. Loss of function studies show that AGE mediated increases in cancer associated processes was dependent upon RAGE expression. Significantly, we show that AGEs are secreted into the tumor microenvironment by cancer cells and may function as signaling molecules to promote immune cell recruitment. These data implicate the AGE-RAGE signaling axis as a potential biological mechanism promoting prostate cancer and may represent a biological mechanism promoting prostate cancer disparity. AGE metabolites may have high potential impact as prognostic/diagnostic markers and/or as a novel area of potential therapeutic intervention to reduce cancer disparity. Citation Format: Danzell Smith, Dion Foster, Van Phan, Victoria Findlay, Lourdes Nogueira, Laura Spruill, Mahtabuddin Ahmed, Judith Salley, Marvella Ford, David Turner. Mechanistic implications of advanced glycation end-products to prostate cancer and racial disparity. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3475.

Tamoxifen is the most widely prescribed adjuvant therapy for estrogen receptor positive (ER+) breast cancer which comprises around 70% of all breast cancer cases. While many patients respond positively to tamoxifen treatment around 50% have de-novo resistance and approximately 30% of responsive tumors recur due to acquired drug resistance. Compared to European American women, African-American women with ER+ breast cancer, have worse progression-free and overall survival which coincides with increased resistance to anti-cancer therapies such as tamoxifen. Poor diet, low income, obesity and a lack of exercise are established lifestyle factors that are known to increase cancer burden and are often more prevalent in African American communities. Our research has identified a potential mechanistic link between sugar derived metabolites and estrogen receptor (ER) phosphorylation which provides a biological consequence of these established lifestyle factors which may directly impact tamoxifen therapy and minority health. Glycation is the non-enzymatic glycosylation of sugar moieties to biological macromolecules such as protein and DNA which produces reactive metabolites known as advanced glycation end products (AGE's). AGE's accumulate in our tissues as we grow older and drive many of the complications associated with diseases displaying health disparity including diabetes, metabolic syndrome, Alzheimer's and heart disease. Significantly, low income, obesity and an inactive lifestyle are established factors driving health disparity that also contribute to increased AGE accumulation levels in our bodies. In particular, AGE content in the Western Diet has consistently increased over the last 50 years due to increased consumption of sugar laden and cheap processed/manufactured foods which are high in reactive AGE metabolites and promote obesity. Due to the established links with lifestyle and cancer disparity we examined the biological effects of AGEs on tamoxifen therapy and found that elevated AGE levels can directly affect the response to tamoxifen treatment and promote tamoxifen resistance. AGE treatment in ER+ breast cancer cell models promoted tamoxifen resistance via the activation of the MAPK and AKT pathways leading to resistance associated changes in ERα phosphorylation. We also observed greater levels of AGE and its cognate receptor for AGE (RAGE) within breast cancer tumor and serum samples and showed a correlation between tumor progression and intra-tumoral AGE concentration. Strategies to eliminate or delay the occurrence of tamoxifen resistance would contribute to increasing overall survival in minority populations. By associating lifestyle-derived AGEs with tamoxifen resistance, opportunities exist for impacting cancer treatment initiatives arising through health and nutritional education and community outreach programs driven by basic, translational, epidemiological and cancer prevention research initiatives. Citation Format: Katherine R. Walter, Danzell M. Smith, Van Phan, Laura spruill, Marvella E. Ford, Victoria J. Findlay, David P. Turner. Defining the implications of sugar derived metabolites (AGEs) to tamoxifen resistance and breast cancer disparity: Is it a question of lifestyle? [abstract]. In: Proceedings of the Eighth AACR Conference on The Science of Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; Nov 13-16, 2015; Atlanta, GA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2016;25(3 Suppl):Abstract nr C65.

Poor diet, low income, obesity and a lack of exercise are established lifestyle factors that are known to increase cancer burden and are often more prevalent in African American communities. As our understanding of tumor biology advances, it is becoming increasingly clear that these inter-related lifestyle factors have distinct molecular consequences on the biological make-up of tumors, altering cell signaling events and gene expression profiles to contribute to cancer disparity outcomes such as its earlier development or its progression to more aggressive disease. Sparse information exists about the genetic and biological factors that contribute to differential cancer survival and mortality rates observed in minority populations. A greater understanding of the interplay between risk factors and the molecular mechanisms associated with cancer disparity will significantly impact minority health. Advanced glycation end products (AGEs) are reactive metabolites produced as a by-product of sugar metabolism. Failure to remove these highly reactive metabolites can lead to protein damage, aberrant cell signaling, increased stress responses, and decreased genetic fidelity. Critically, AGE accumulation is also directly affected by our lifestyle choices such as poor diet, low income, obesity and a lack of exercise. We recently reported a potential mechanistic link between AGEs and prostate cancer which may provide a molecular consequence of our lifestyle choices that can directly impact tumor biology and contribute to cancer disparity. We examined circulating and intra-tumoral AGE metabolite levels in clinical specimens and identified a race specific, tumor dependent pattern of accumulation in prostate cancer serum and tumor. Further mechanistic studies in immortalized prostate cancer cell lines show that AGE treatment increases the expression of the receptor for AGEs (RAGE) to activate cancer-associated signaling cascades. Loss of function studies show that AGE mediated increases in cancer associated processes was dependent upon RAGE expression. Significantly, we show that AGEs are secreted into the tumor microenvironment by cancer cells and may function as signaling molecules to promote immune cell recruitment. These data implicate the AGE-RAGE signaling axis as a potential biological mechanism promoting prostate cancer and may represent a biological mechanism promoting prostate cancer disparity. AGE metabolites may have high potential impact as prognostic/diagnostic markers and/or as a novel area of potential therapeutic intervention to reduce cancer disparity. Citation Format: Danzell M. Smith, Dion Foster, Van Phan, Victoria J. Findlay, Lourdes M. Nogueira, Laura Spruill, Mahtabuddin Ahmed, Judith D. Salley, Marvella E. Ford, David P. Turner. Mechanistic implications of advanced glycation end-products to prostate cancer and racial disparity. [abstract]. In: Proceedings of the Eighth AACR Conference on The Science of Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; Nov 13-16, 2015; Atlanta, GA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2016;25(3 Suppl):Abstract nr C76.

Past studies have identified that a unique type of matrix metalloproteinase, the membrane-type-1 MMP (MT1-MMP), is increased within the left ventricle (LV) of patients with dilated cardiomyopathy (DCM). However, the cellular and molecular basis for this induction of MT1-MMP with DCM is unknown. LV myocardial biopsies from nonfailing, reference normal patients (defined as LV ejection fraction >50%, elective coronary bypass surgery, no perfusion defect at biopsy site, n = 6) and DCM patients (LV ejection fraction

African American cancer patients are more likely to die of their disease than their European counterparts. Our research has identified a potential mechanistic link between carbohydrate derived metabolites and cancer associated pathways which may provide a biological consequence of the socioeconomic and environmental factors that are known to drive cancer health disparity. Glycation is the non-enzymatic glycosylation of sugar moieties to biological macromolecules such as protein and DNA which produces reactive metabolites known as advanced glycation end products (AGE's). AGE's accumulate in our tissues as we grow older and drive many of the complications associated with diseases displaying health disparity including diabetes, metabolic syndrome, Alzheimer's and heart disease. Low income, obesity and an inactive/sedentary lifestyle are established factors driving cancer health disparity. Apart from their production during normal metabolism, AGE's are also formed through the ingestion of food and by external environmental factors such as lack of exercise. AGE content in the Western Diet has consistently increased over the last 50 years due to increased consumption of sugar laden and cheap processed/manufactured foods which are high in reactive AGE metabolites and can promote obesity. We therefore examined circulating and intra-tumoral AGE metabolite levels in clinical specimens and identified a race specific, tumor dependent pattern of accumulation in prostate cancer serum and tumor. In mouse xenograft models, AGE accumulation was highest in the more aggressive tumors. One way AGE's mediate their deleterious effects is by functioning as ligand for the trans-membrane receptor for AGE (RAGE). In diabetes and other diseases, the AGE-RAGE signaling axis is a pro-inflammatory pathway leading to the upregulation of pro-inflammtory cytokines through increased NFkB activation. Higher AGE levels in prostate tumors corresponded with higher RAGE expression and increased NFkB transcriptional activity. In immortalized prostate cancer cell lines AGE treatment increased cancer associated processes and RAGE expression levels. Loss of function studies show that AGE mediated increases in cancer associated processes was dependent upon RAGE expression. These data implicate the AGE-RAGE signaling axis as a potential biological mechanism promoting prostate cancer and indicate that increased AGE accumulation may represent a biological mechanism promoting prostate cancer disparity. This abstract is also presented as Poster C62. Citation Format: David P. Turner, Victoria J. Findlay, Lourdes M. Nogueira, Sadia Robinson, Emily Kistner-Griffin, Laura Spruill, Ryan Y. Turner, Mahtabuddin Ahmed, Judith D. Salley, Marvella E. Ford. Advanced glycation end-products are increased in prostate cancer and may promote racial disparity. [abstract]. In: Proceedings of the Sixth AACR Conference: The Science of Cancer Health Disparities; Dec 6–9, 2013; Atlanta, GA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2014;23(11 Suppl):Abstract nr PR10. doi:10.1158/1538-7755.DISP13-PR10