Presence Of Crown-like Structures Research Articles

Presence of crown-like structures in breast adipose tissue; differences between healthy controls, BRCA1/2 gene mutation carriers and breast cancer patients.

Crown-like structures (CLS) in breast adipose tissue are associated with inflammation and a potential factor in breast cancer behaviour. Whether this effect varies between breast cancer subtypes and is influenced by BMI and BRCA mutation status is presently unknown. Therefore, we compared CLS presence between adipose tissue of healthy controls, BRCA1/2 gene mutation carriers and breast cancer patients, and assessed the relation of CLS with clinical outcome in breast cancer patients. Immunohistochemical staining for CD68 was performed on breast adipose tissue sections of 48 healthy controls, 78 BRCA1/2 gene mutation carriers and 259 breast cancer patients. CLS presence and index (CLS/cm2) were correlated with BMI, BRCA status, tumour presence, intrinsic tumour subtype and tumour characteristics. Associations with clinical outcome were assessed. CLS were more often present in breast cancer patients compared to BRCA carriers and healthy controls. CLS presence was associated with the presence of breast cancer and high BMI. CLS were more often present in Luminal-B-like tumours compared to the other subtypes. No correlations between CLS and BRCA status or age was found. In TNBC, CLS were related to lymphovascular invasion. No association with survival was found. In conclusion, CLS were more frequently present in breast adipose tissue of breast cancer patients compared to BRCA1/2 gene mutation carriers and healthy controls. Furthermore, our study provides evidence of the association between obesity and presence of CLS. The prognostic significance and impact on clinical outcome of differences in CLS numbers should be further assessed in prospective studies.

Prevalence and patterns of breast white adipose tissue inflammation among American Black women and indigenous Nigerian women with early-stage breast cancer.

e12560 Background: Racial disparities in breast cancer outcomes pose a major public health challenge. Complex interactions between social determinants of health and tumor biology, mediated by obesity and adiposity, may play an important role in these disparities. White adipose tissue inflammation (WATi) of the breast, which is histologically defined by the presence of crown-like structures (CLS-B), occurs in the setting of excess adiposity and promotes tumor growth via elevated levels of aromatase, proinflammatory growth factors, and metabolic dysregulation. Here we examined relationships between race, body mass index (BMI), and WATi in two cohorts of patients with early-stage breast cancer: 1) American Black women and 2) indigenous Nigerian women. Methods: Non-tumor breast WAT specimens were collected from patients undergoing mastectomy for treatment or prevention of breast cancer at Memorial Sloan Kettering Cancer Center (MSK) in the US and Obafemi Awolowo University Teaching Hospital (OAUTH) in Nigeria. WATi was defined by the presence of CLS-B as detected by CD68 immunostaining. All tissue analyses were performed at MSK. Associations among BMI and WATi were tested within each cohort using Kendall’s tau rank correlation and compared between cohorts using Pearson's Chi-squared test. Results: A total of 224 American women were enrolled at MSK between 2011-2018, of whom 39 self-reported Black race. At OAUTH, 97 Nigerian women were enrolled between 2018-2022. Invasive carcinoma was present in 32 American Blacks and 97 Nigerians. Of those with invasive carcinoma, the prevalence of WATi among American Black women was 72% vs. 37% in the Nigerian cohort ( p<0.001). In the US cohort, there was no significant difference between the prevalence of WATi among Black vs. non-Black patients (72% vs. 62%; p=0.3). 44% of American Blacks were obese and 34% overweight compared with 27% and 39% of Nigerians, respectively (p=0.07). The prevalence of WATi varied by BMI category within each cohort, as shown. Overweight/obesity was associated with WATi in the Nigerian (p=0.009) and American cohorts (p<0.001); a trend was observed in the American Black subgroup (p=0.13). Conclusions: Breast WATi and obesity were more prevalent among American Black women with breast cancer compared with Nigerian women. These findings highlight the need for further investigation of environment, lifestyle patterns, and body composition differences between indigenous and diaspora populations. [Table: see text]

Abstract P5-03-08: Joint associations of mammographic breast density and obesity on the presence of crown-like structures in the breast adipose tissue of breast cancer patients

Abstract Background: Obesity is an established risk factor for postmenopausal breast cancer and is associated with poor outcomes. Accumulating evidence suggests crown-like structures in the breast adipose tissue (CLS-B), a marker of local inflammation, play a role in explaining the obesity-breast cancer association. However, it is unknown whether breast tissue composition (i.e., the amount of fibroglandular tissue in the breast relative to fat) is related to CLS-B. Objective: We evaluated whether breast tissue composition, as reflected by mammographic breast density, is associated with breast adipose tissue inflammation, as indicated by the presence of CLS-B, and whether the combination of breast density and obesity increases the presence of CLS-B among newly diagnosed breast cancer patients. Methods: We examined the presence of CLS-B, detected by CD68 immunohistochemistry, in breast adipose tissue obtained via mastectomy from a quadrant uninvolved by tumor among 254 women with stage I–III breast cancer treated at Emory University Hospitals (2007–2012). Patient characteristics, including mammographic breast density (assessed on a mammogram up to 5 years before breast surgery) and body mass index (BMI) at diagnosis, were abstracted from electronic medical records. Mammographic density was assessed using the Breast Imaging Reporting and Data System (BI-RADS) density classification (1=almost entirely fat; 2=scattered fibroglandular densities; 3=heterogeneously dense; and 4=extremely dense); density was further categorized as fatty (BI-RADS 1-2) and dense (BI-RADS 3-4). Age and multivariable (MV)-adjusted logistic regression models were used to estimate odds ratios (ORs) and 95% confidence intervals (CIs) for the independent and joint associations of breast density (dense vs. fatty) and obesity (BMI ≥30 kg/m2 vs. < 30 kg/m2) on the presence of CLS-B. Multivariable models adjusted for age (continuous, years) and parity (nulliparous, parous) with mutual adjustment of BMI (continuous, kg/m2) or mammographic breast density (dense, fatty) depending on the model. Results: Women with obesity were more likely to have fatty breasts than women without obesity (52% vs. 27%). Obesity was strongly associated with the presence of CLS-B in age-adjusted (OR=3.11, 95% CI: 1.79, 5.48) and multivariable models (MV-OR=3.70, 95% CI: 1.96, 7.15). There was no apparent association between dense breast tissue and presence of CLS-B in the age-adjusted model (OR=1.04, 95% CI: 0.59, 1.85). After additional adjustment for BMI and parity, we noted that women with dense breasts had higher odds of having CLS-B compared to those with fatty breasts (MV-OR=2.13, 95% CI: 1.07, 4.41). MV-ORs from the joint model (common referent: not obese, fatty breasts) were 1.54 (95% CI: 0.62, 4.24) for women without obesity but with dense breasts, 3.18 (95% CI: 1.15, 9.56) for women with obesity and fatty breasts, and 6.24 (95% CI: 2.23, 19.2) for women with obesity and dense breasts. Conclusions: Our findings suggest that dense rather than fatty breast tissue is associated with breast adipose tissue inflammation among women with breast cancer. Results of the joint analyses suggest obesity is more strongly predictive of CLS-B presence than breast density. However, density may be an important risk factor for CLS-B among women without obesity while patients with obesity and dense breast tissue are the most likely to have CLS-B present. Future studies may consider the mechanisms by which density leads to increased presence of CLS-B. Joint Associations Table Table 1. Joint associations between mammographic breast density (MD) and body mass index (BMI) on the presence of crown-like structures in breast adipose tissue (CLS-B) among 254 women diagnosed with invasive breast cancel (stage I-III) at Emory University between 2007 and 2012 Citation Format: Maret L. Maliniak, Rebecca L. Seidel, Kimberly Bertrand, Lauren E. McCullough. Joint associations of mammographic breast density and obesity on the presence of crown-like structures in the breast adipose tissue of breast cancer patients [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P5-03-08.

Abstract C074: Presence of crown-like structures in breast adipose tissue and clinical outcomes among African-American and White breast cancer patients

Abstract Background: Crown-like structures in breast adipose tissue (CLS-B), indicative of proinflammatory conditions, are most frequently observed among obese (body mass index, BMI ≥30 kg/m2) women and may contribute to poor prognosis in this group. African-American (AA) women have disproportionately higher rates of obesity than White women, and at least one prior study suggests the prevalence of CLS-B may be higher among AA women. However, most previous studies have examined CLS-B within affected tissues, which may reflect inflammation in the tumor microenvironment, and few have examined the association between CLS-B and clinical outcomes by race. Methods: We examined the presence of CLS-B detected by CD68 immunohistochemistry in normal adjacent breast tissue from a quadrant uninvolved by tumor obtained via mastectomy among 174 African-American women and 168 White women with stage I—III breast cancer diagnosed at Emory University Hospitals (2007—2012). We also investigated associations between CLS-B and other demographic and lifestyle factors at diagnosis (e.g., BMI, smoking status, age at menarche, parity, lactation, menopausal status, hormone replacement therapy use, and family history of breast cancer). Patients were followed for an average of seven years after diagnosis for recurrence and survival. Multivariable Cox proportional hazards models were used to compute hazard ratios (HR) and 95% confidence intervals (CI) for associations between CLS-B presence and progression-free survival (PFS), controlling for BMI and other potential confounders. Results: Median age at diagnosis for both AA and White women was 54 years, with more than 60% postmenopausal among both groups. AA women were more likely than White women to be obese (52% vs. 24%) and have ER- tumors (30% vs. 12%). Presence of any CLS-B was similar between AA (32%) and White (29%) patients. In multivariable models, we did not find any association between CLS-B and race (HR=1.14, 95% CI: 0.72, 1.82) with the only statistically significant factors being BMI (≥30 vs. 18.5-<25 kg/m2: HR=4.36, 95% CI: 2.17, 8.76) and parity (1+ vs. 0 births: HR=0.43, 95% CI: 0.21, 0.91). Over follow-up, 46 breast cancer recurrences and 52 deaths (23 from breast cancer) occurred. Overall, the presence of CLS-B was not associated with PFS (multivariable HR: 0.97, 95% CI: 0.58, 1.62). When examined by race, there was a difference in the direction of the association between CLS-B and PFS among AA women (HR=1.25, 95% CI: 0.64, 2.46) compared to White women (HR=0.75, 95% CI: 0.33, 1.71), although this difference was not statistically significant (P=0.86). Conclusion: Our results show a strong, positive association between BMI and CLS-B in non-tumor tissue and an inverse association with parity. We did not observe a difference in CLS-B presence by race nor did we find CLS-B to be associated with worse progression-free survival, which is in contrast to previous studies that have examined the presence of CLS-B within specimens in close proximity to the tumor. Citation Format: Aswathy M Cheriyan, Mark E Sherman, Yuan Liu, Keerthi Gogineni, Jiaqi Liu, Jiabei He, Uma Krishnamurti, Ryan Ashiqueali, Jinjing He, Rami Yacoub, Jasmine Miller-Kleinhenz, Lauren E McCullough, Maret L Maliniak. Presence of crown-like structures in breast adipose tissue and clinical outcomes among African-American and White breast cancer patients [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 C074.

1749-P: The TNFa Inhibitor Infliximab Decreases Inflammation in High-Fat Fed Mice but Does Not Alleviate Insulin Resistance

The insulin resistance of diet-induced obesity (DIO) is accompanied by inflammation. Previous studies suggest that the anti-inflammatory effects of infliximab, an anti-TNFα chimeric monoclonal antibody, decreases DIO-associated inflammation in mice fed a 60% fat diet. The effectiveness of infliximab at 10 mg/kg Infliximab (IF10) and 100 mg/kg infliximab (IF100) in C57Bl/6J DIO mice was studied in comparison to C57Bl/6J DIO mice administered chimeric IgG (IgG; 10 mg/kg). Treatment consisted of IP injections of infliximab or IgG once per week for 4 weeks. Body weight and fasting arterial glucose was not different in IF10, IF100, and IgG. IF10 and IF100 mice had a decreased presence of crown-like structures in white adipose tissue compared to IgG treated mice. Infliximab resulted in a dose-dependent reduction in macrophage F4/80 staining in white adipose tissue (4.3±0.7% and 1.8±0.2% in IF10 and IF100, respectively) compared to 8.1±1.6% in IgG, validating the anti-inflammatory effect. Insulin action was measured in mice with arterial and venous catheters using the hyperinsulinemic, euglycemic clamp method (insulin dose of 4 mU/kg/min) combined with [3-3H]glucose in conscious, unstressed mice. Glucose infusion rates (GIR) in IgG mice were higher (25±3 mg/kg/min) as compared to IF10 (17±2 mg/kg/min) and IF100 (19±2 mg/kg/min). The greater GIR in IgG as compared to IF10 was due to changes in hepatic endogenous glucose production (suppression of 12±3 mg/kg/min and 5±1 mg/kg/min, respectively) without a difference in glucose utilization. IF100 exhibited small reductions in both hepatic and peripheral insulin action, neither of which reached significance. Mice treated with saline alone were used as an additional control, but neither IF10 nor IF100 had significant effects on insulin action compared to saline controls. In summary, despite profound anti-inflammatory effects of the anti-TNFα drug, Infliximab causes a paradoxically greater hepatic insulin resistance. Disclosure D.A. Roby: None. C. Guan: None. F. James: None. D. Bracy: None. L. Lantier: None. D. Wasserman: None.

Reduction of IL-6 gene expression in human adipose tissue after sleeve gastrectomy surgery.

SummaryObjectiveThere is increasing evidence that immune cell interactions in adipose tissue contribute to the development of metabolic dysfunction. Pro‐inflammatory cytokines have been shown to mediate insulin resistance, and the presence of macrophages is a salient feature in the development of obesity. The present study aimed to evaluate adipocyte size and macrophage activation in women before and 3 months after laparoscopic vertical sleeve gastrectomy (VSG).MethodsSubcutaneous abdominal adipose tissue biopsies were obtained from women scheduled to undergo VSG. Histological evaluation of adipocytes and macrophages was performed as well as cytokine expression quantification before and after VSG‐induced weight loss.ResultsWeight loss following VSG resulted in a reduction in adipocyte size as well as a decrease in interleukin (IL)‐6 cytokine mRNA expression in subcutaneous adipose tissue. There was no change in the presence of crownlike structures after weight loss.ConclusionsEarly weight loss after VSG is associated with a reduction in adipocyte size and a decline in IL‐6 gene expression in local adipose tissue. Macrophage infiltration and crownlike density structures persist in adipose tissue from tissues impacted by excess body weight 3 months after VSG‐induced weight loss.

Improving risk assessment of obesity-associated breast cancer.

1544 Background: Elevated body mass index (BMI) is associated with increased risk of estrogen receptor (ER)-positive postmenopausal breast cancer. The risk is also elevated in women with a normal BMI but excess body fat. These risks may be driven by breast white adipose tissue inflammation (WATi), which is associated with elevated aromatase levels and systemic metabolic dysfunction (e.g. hyperinsulinemia). We hypothesized that body fat assessment is superior to BMI for detecting the pathophysiology that promotes obesity-related breast cancer, particularly among normal BMI women. Methods: Non-tumorous breast tissue was collected from women undergoing mastectomy for breast cancer treatment or prevention. Breast WATi was detected by the presence of crown-like structures in the breast, which are composed of a dead/dying adipocyte surrounded by CD68+ macrophages. Body composition was measured prior to mastectomy via dual energy X-ray absorptiometry. Exercise behavior was also assessed prior to surgery using the Godin Leisure Time Exercise Questionnaire. Associations among categorical variables were examined using Χ2 or Fisher’s exact test. Relationships between continuous variables were examined using the Spearman correlation. Results: From April 5, 2016 to August 31, 2018, 100 patients were enrolled; median age 49 (range 29 to 82) years. Breast WATi was present in 56/100 (56%) women and was associated with elevated BMI and body fat levels, breast adipocyte hypertrophy, postmenopausal status, metabolic syndrome and decreased physical activity (P < 0.05). Among 39 women with normal BMI, breast WATi was present in 14 (36%) and was associated with elevated body fat levels, breast adipocyte hypertrophy, dyslipidemia, and decreased physical activity (P < 0.05). There was no statistically significant association between BMI and breast WATi in the normal BMI group. Menopausal status and total fat mass had greater sensitivity and specificity for the detection of breast WATi compared to a BMI-based model (AUC 0.843 vs. 0.779, respectively). Conclusions: Measurement of body fat is superior to BMI for predicting breast inflammation, which has been shown to promote obesity-related breast cancer.

Abstract P5-07-05: Obesity, adipose inflammation, and race in patients with early stage breast cancer

Abstract Background: Elevated body mass index (BMI) is associated with increased risk of estrogen receptor-positive postmenopausal breast cancer. Mechanistically, most individuals with elevated BMI have breast white adipose tissue inflammation (WATi) which confers increased breast cancer risk, particularly in those with existing benign breast disease. Individuals with WATi have elevated in-breast expression of aromatase and several systemic changes that increase breast cancer risk, including hyperinsulinemia and higher levels of C-reactive protein. However, women with normal BMI but high levels of body fat are also likely to harbor WATi and are at increased risk of postmenopausal breast cancer. The accuracy of BMI for assessing adiposity and predicting obesity-related disorders, including cancer, varies across race and ethnicity. Whether the association between BMI and WATi varies by race is unknown. Here we aimed to characterize relationships among breast WATi and clinicopathologic features in a racially diverse cohort undergoing mastectomy for breast cancer treatment. Methods: Non-tumorous breast tissue and fasting blood were collected from women undergoing mastectomy for breast cancer treatment or prevention at a single center serving a racially diverse patient population. Breast WATi was detected by the presence of crown-like structures in the breast (CLS-B), which are composed of a dead/dying adipocyte surrounded by CD68+ macrophages. Clinicopathologic data were abstracted from electronic medical records. Associations among categorical variables were examined using Fisher's exact test. Relationships between continuous variables were examined using the Spearman correlation. Results: As of May 18, 2018 62 patients have been accrued; median age 55 (range 32 to 84). Self-reported race distribution was: 36 (58%) Asian, 5 African American (8%), 20 (32%) Caucasian, and 1 (2%) unknown. Breast tissue has been analyzed for WATi in 60 cases thus far. Clinicopathologic features stratified by the presence or absence of breast WATi are presented in. Breast WAT inflammation was associated with obesity (P=0.02) and a trend to association was observed with dyslipidemia (P<0.09). VariableBreast WATi Absent (n=25)Breast WATi Present (n=35)Age, years Median (range)51 (32 to 71)59 (36 to 80)BMI, kg/m2 Median (range)22.5 (18.1 to 35.3)28.0 (19.2 to 38.9)BMI Category Underweight1 (4%)0 (0%)Normal16 (64%)10 (29%)Overweight5 (20%)18 (51%)Obese3 (12%)7 (20%)Race, n (%) Asian15 (60%)19 (54%)African American2 (8%)3 (9%)Caucasian8 (32%)12 (34%)Unknown0 (0%)1 (3%)Menopausal Status, n (%) Pre10 (40%)12 (34%)Post15 (60%)23 (66%)Hypertension, n (%)7 (28%)16 (46%)Diabetes mellitus, n (%)0 (0%)5 (14%)Dyslipidemia, n (%)1 (4%)11 (31%) Conclusions: Breast adipose inflammation is associated with elevated BMI and possibly metabolic syndrome disorders in a racially diverse population. These findings are consistent with observations from predominantly Caucasian cohorts. Race-specific characteristics will also be examined. Study accrual is ongoing and updated results will be presented. Citation Format: Iyengar NM, Siegel B, Malik M, Giri DD, Tsai J, Hughes M, Adam A, Williams S, Zhou XK, Rodgers W, Ginter P, Patel A, Yong F, Cherian A, August P, Dannenberg AJ. Obesity, adipose inflammation, and race in patients with early stage breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P5-07-05.

Inflammation in the subcutaneous adipose tissue does not attenuate endothelial function in subjects with diabetes mellitus and subjects with dyslipidaemia and hypertension: Across-sectional study.

SummaryBackgroundObesity is associated with low‐grade inflammation that may be related to vascular disease. We hypothesized that inflammation in the subcutaneous adipose tissue is associated with impaired endothelium‐dependent vasodilatation.MethodsWe assessed endothelial function by measuring forearm vascular response to acetylcholine and determined inflammation in subcutaneous fat biopsies in 2 groups of subjects; 15 patients with type 2 diabetes mellitus (T2DM) and 19 subjects with dyslipidaemia combined with hypertension (DcH). The adipose tissue inflammation score was based on adipocyte size, influx of macrophages and presence of crown‐like structures. We compared the vascular response to acetylcholine between subjects with and without adipose tissue inflammation.ResultsPatients with diabetes had clearly decreased vasodilatation compared to patients with DcH. In total, 23 of the 34 fulfilled the criteria of subcutaneous adipose tissue inflammation. However, there was no difference in vascular response to acetylcholine between the group with and without inflammation (changes in FBF from baseline 3.9 ± 0.8, 7.8 ± 1.0 and 13.6 ± 1.0 mL/dL/min compared to 4.3 ± 1.0, 7.9 ± 2.1 and 12.2 ± 2.4 mL/dL/min in response to acetylcholine 0.5, 2.0 and 8.0 μg/dL/min), nor was there a relationship between systemic hs‐CRP levels and endothelial function.ConclusionsWe confirm that subjects with T2DM have impaired endothelial function compared to age‐ and BMI‐matched subjects with DcH. However, endothelial function did not differ between participants with or without inflammation in the subcutaneous adipose tissue. These results suggest that fat tissue inflammation, at least in the subcutaneous compartment, does not affect vascular function.

High fat diet consumption differentially affects adipose tissue inflammation and adipocyte size in obesity-prone and obesity-resistant rats.

Background/ObjectivesExpanding visceral adiposity is associated with increased inflammation and increased risk for developing obesity-related comorbidities. The goal of this study was to examine high fat diet (HFD)-induced differences in adipocyte size and cytokine/chemokine expression in visceral and subcutaneous adipose depots in obesity-prone (OP) and obesity-resistant (OR) rats.MethodsOP and OR rats were fed either a low fat diet (LFD, 10% kilocalories from fat) or HFD (60% kilocalories from fat) for 7 weeks. Adipocyte size and the presence of crown-like structures in epididymal and inguinal adipose tissue were determined. A multiplex cytokine/chemokine panel was used to assess the expression of inflammatory markers in epididymal and inguinal adipose tissues.ResultsA higher percentage of large adipocytes (> 5000 μm2) was detected in the epididymal and inguinal adipose tissues of OP rats and a higher percentage of small adipocytes (< 4000 μm2) was detected in the epididymal and inguinal adipose tissues of OR rats. More crown-like structures were identified in epididymal adipose tissue of OP rats fed a LFD, compared to OR rats. Consumption of a HFD increased the number of crown-like structures in OR, but not OP rats. Epididymal expression of pro-inflammatory cytokines (IL-1β, TNF-α) was higher in OP rats, compared to OR rats fed LFD. HFD consumption increased epididymal expression of GM-CSF, IL-1α, IL-1β, IL-6, MIP-2, and TNF-α in OP and OR rats. Inguinal expression of pro-inflammatory cytokines (IL-1α, IL-1β, TNF-α) was higher in OP rats, compared to OR rats.ConclusionsOverall, these data suggest that a higher susceptibility to developing obesity is characterized by large adipocytes and increased visceral adipose inflammation. Interestingly, in OR rats, the detrimental effects of HFD consumption on visceral adipose inflammation are evident with only small increases in weight and adiposity, suggesting that HFD also increases the risk for obesity-related comorbidities in OR rats.

Abstract 1251: Pioglitazone inhibits periprostatic white adipose tissue inflammation in obese mice

Abstract Obesity is associated with an increased incidence of high-grade prostate cancer (PC) and poor prognosis for PC patients. Recently, we showed that obesity-related periprostatic white adipose tissue (WAT) inflammation, characterized by crown-like structures (CLS) consisting of dead or dying adipocytes surrounded by macrophages, was associated with high-grade PC in men. It’s possible, therefore, that agents that suppress periprostatic WAT inflammation will alter the natural history of PC. Pioglitazone, a ligand of PPARγ, is used to treat diabetes and possesses anti-inflammatory properties. Here our main objectives were to determine if pioglitazone inhibited obesity-related periprostatic WAT inflammation in mice and then to elucidate the underlying mechanism. Mice were fed either a high fat (HF) diet or low fat (LF) diet to determine if obesity caused periprostatic WAT inflammation as defined by the presence of CLS. To determine if pioglitazone inhibited obesity-associated periprostatic WAT inflammation, obese HF diet fed mice were either continued on HF diet alone or treated with HF diet containing two doses (0.005% or 0.05% w/w) of pioglitazone. Treatment with pioglitazone reduced the density of CLS in periprostatic WAT, and suppressed levels of TNF-α, TGF-β and the chemokine monocyte chemoattractant protein-1 (MCP-1). Importantly, the ability of pioglitazone to suppress periprostatic WAT inflammation was abrogated in MCP-1 knock out mice. Pioglitazone caused dose-dependent induction of both adiponectin, an anti-inflammatory adipokine, and its receptor AdipoR2 in cultured 3T3-L1 cells and in periprostatic WAT of obese mice. Pioglitazone blocked TNF-α-mediated induction of MCP-1 in 3T3-L1 cells, an effect that was attenuated when either adiponectin or AdipoR2 were silenced. Taken together, pioglitazone-mediated induction of adiponectin suppressed the elevation in MCP-1 levels thereby attenuating obesity-related periprostatic WAT inflammation. These findings strengthen the rationale for future efforts to determine whether targeting the PPARγ-adiponectin-MCP-1 axis will decrease periprostatic adipose inflammation and thereby reduce the risk of high-grade PC or improve outcomes for men with PC. Citation Format: Miki Miyazawa, Kotha Subbaramaiah, Priya Bhardwaj, Xi Kathy Zhou, Hanhan Wang, Domenick J. Falcone, Dilip D. Giri, Andrew J. Dannenberg. Pioglitazone inhibits periprostatic white adipose tissue inflammation in obese mice [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 1251. doi:10.1158/1538-7445.AM2017-1251

Periprostatic adipose inflammation is associated with high-grade prostate cancer.

BackgroundObesity, a cause of subclinical inflammation, is associated with increased risk of high grade prostate cancer (PC) and poor outcomes. Whether inflammation occurs in periprostatic white adipose tissue (WAT), and contributes to the negative impact of obesity on PC aggressiveness, is unknown.MethodsIn a single-center, cross-sectional design, men with newly diagnosed PC undergoing radical prostatectomy were eligible for study participation. The primary objective was to examine the prevalence of periprostatic WAT inflammation defined by the presence of crown-like structures (CLS-P) as detected by CD68 immunohistochemistry. Secondary objectives were to explore the clinical and systemic correlates of periprostatic WAT inflammation. Tumor characteristics and host factors including BMI, adipocyte diameter, and circulating levels of lipids, adipokines, and other metabolic factors were measured. Wilcoxon rank-sum, Chi-square, or Fisher’s exact tests, and generalized linear regression were used to examine the association between WAT inflammation and tumor and host characteristics.ResultsPeriprostatic fat was collected from 169 men (median age 62 years; median BMI 28.3). Periprostatic WAT inflammation was identified in 49.7% of patients and associated with higher BMI (P=0.02), larger adipocyte size (P=0.004), and Gleason grade groups IV/V tumors (P=0.02). The relationship between WAT inflammation and high Gleason grade remained significant after adjusting for BMI (P=0.04). WAT inflammation correlated with higher circulating levels of insulin, triglycerides, and leptin/adiponectin ratio, and lower high density lipoprotein cholesterol, compared to those without WAT inflammation (P’s <0.05).ConclusionsPeriprostatic WAT inflammation is common in this cohort of men with PC and is associated with high grade PC.

Effect of bariatric surgery on breast tissue and biomarkers in obese women at increased risk for breast cancer.

TPS1592 Background: Obesity represents a challenging epidemic associated with increased risk of several malignancies, including breast cancer in post-menopausal women. Proposed mechanisms for the association between obesity and breast cancer risk include increased insulin resistance, elevated levels of circulating estrogens, and chronic inflammation. Intentional weight loss from bariatric surgery has been associated with decreased risk of breast cancer. While rapid improvements in serologic markers of metabolism and inflammation are seen following bariatric surgery, short- and long- term changes in breast tissue remain less clear. This study investigates the effect of bariatric surgery on breast density and biomarkers of increased risk in breast tissue. Methods: This pilot, single institution, observational study (NCT02681120) is recruiting pre- and post-menopausal women with BMI ≥30 from a University bariatric surgery clinic using the Hughes risk application as a screening tool. Eligible patients must have a lifetime risk for breast cancer of ≥20%. Participants are evaluated by imaging, breast biopsy, and blood samples at baseline, 14 days post-operatively to determine the effects of rapid metabolic changes, and 1 year post-operatively to determine the effects of significant weight loss. The impact of bariatric surgery on known imaging parameters of breast cancer risk is assessed by background parenchymal enhancement on MRI and breast density on mammogram. Breast tissue is evaluated for changes in immune infiltrates, aromatase expression, and the presence of crown-like structures, a marker of inflammation seen in the breast tissue of obese women. Tissue samples at each time point are also compared to samples from lean women in the Susan G. Komen Tissue Bank at the IU Simon Cancer Center. Blood is collected for correlative studies evaluating markers of inflammation, insulin resistance, metabolism, and hormone synthesis. Enrollment is currently ongoing with a planned accrual of 40 patients, and data collection is estimated to complete by the end of 2018. Clinical trial information: NCT02681120.

Inflammation of mammary adipose tissue occurs in overweight and obese patients exhibiting early-stage breast cancer

Growing evidence indicates that adiposity is associated with breast cancer risk and negatively affects breast cancer recurrence and survival, a paracrine role of mammary adipose tissue being very likely in this process. In contrast to other adipose depots, occurrence of a sub-inflammatory state of mammary adipose tissue defined by dying adipocytes surrounded by macrophages forming crown-like structures in overweight and obese subjects, remains only partially described. In a general population of breast cancer patients (107 patients) mostly undergoing breast-conserving surgery, we found a positive association between patient’s body composition, breast adipocytes size, and presence of crown-like structures in mammary adipose tissue close to the tumor. Overweight (BMI: 25.0–29.9 kg/m2) and obese (BMI ≥ 30.0 kg/m2) patients have 3.2 and 6.9 times higher odds ratio of crown-like structures respectively, compared with normal weight patients. The relatively small increase in adipocyte size in crown-like structures positive vs. negative patients suggests that mammary adipose tissue inflammation might occur early during hypertrophy. Our results further highlight that body mass index is an adequate predictor of the presence of crown-like structures in mammary adipose tissue among postmenopausal women, whereas in premenopausal women truncal fat percentage might be more predictive, suggesting that mammary adipose tissue inflammation is more likely to occur in patients exhibiting visceral obesity. Finally, the presence of crown-like structures was positively associated with systemic markers such as the Triglyceride/High-density lipoprotein-cholesterol ratio serum C-reactive protein and glucose/(HbA1c) glycated Haemoglobin. These compelling results demonstrate that excess adiposity, even in overweight patients, is associated with mammary adipose tissue inflammation, an event that could contribute to breast cancer development and progression.

Effects of Rapid Weight Loss on Systemic and Adipose Tissue Inflammation and Metabolism in Obese Postmenopausal Women.

Context:Obesity is associated with subclinical white adipose tissue inflammation, as defined by the presence of crown-like structures (CLSs) consisting of dead or dying adipocytes encircled by macrophages. In humans, bariatric surgery-induced weight loss leads to a decrease in CLSs, but the effects of rapid diet-induced weight loss on CLSs and metabolism are unclear.Objective:To determine the effects of rapid very-low-calorie diet-induced weight loss on CLS density, systemic biomarkers of inflammation, and metabolism in obese postmenopausal women.Design:Prospective cohort study.Setting:Rockefeller University Hospital, New York, NY.Participants:Ten obese, postmenopausal women with a mean age of 60.6 years (standard deviation, ±3.6 years).Main Outcome Measures:Effects on CLS density and gene expression in abdominal subcutaneous adipose tissue, cardiometabolic risk factors, white blood count, circulating metabolites, and oxidative stress (urinary isoprostane-M) were measured.Results:Obese subjects lost approximately 10% body weight over a mean of 46 days. CLS density increased in subcutaneous adipose tissue without an associated increase in proinflammatory gene expression. Weight loss was accompanied by decreased fasting blood levels of high-sensitivity C-reactive protein, glucose, lactate, and kynurenine, and increased circulating levels of free fatty acids, glycerol, β-hydroxybutyrate, and 25 hydroxyvitamin D. Levels of urinary isoprostane-M declined.Conclusion:Rapid weight loss stimulated lipolysis and an increase in CLS density in subcutaneous adipose tissue in association with changes in levels of circulating metabolites, and improved systemic biomarkers of inflammation and insulin resistance. The observed change in levels of metabolites (i.e., lactate, β-hydroxybutyrate, 25 hydroxyvitamin D) may contribute to the anti-inflammatory effect of rapid weight loss.

Incidence of periprostatic white adipose tissue inflammation in men with prostate cancer.

63 Background: Obesity, a common cause of chronic inflammation, is associated with an increased risk of high grade, lethal prostate cancer (PC) and poor outcomes. The existence or clinical importance of periprostatic white adipose tissue inflammation (WATi) in patients (pts) with PC has not been previously described. We examined the relationships among periprostatic WATi and 1) tumor clinicopathologic features, and 2) host factors including age, body mass index (BMI), and circulating metabolic factors. Methods: Periprostatic WAT was collected prospectively from men with PC undergoing radical prostatectomy. WATi was defined by the presence of dead/dying adipocytes surrounded by macrophages forming crown-like structures (CLS). Tumor characteristics and host factors were measured. Wilcoxon rank-sum, Chi-square, or Fisher’s exact tests were used to examine the relationship between WATi and tumor and host characteristics. Results: From 11/2011-8/2015, periprostatic WAT was obtained from 169 pts (median age 62 years, range: 39 -77). Fasting blood samples were collected from 154 pts. CLS were present in 84 (49.7%) of pts. Presence of CLS was associated with higher median BMI (P = 0.02); 40/65 (61.5%) obese pts, 36/83 (43.4 %) overweight pts, and 8/21 (38.1 %) normal weight pts had CLS. Pts with CLS were more likely to have high grade prostate cancer (Gleason grade group IV/V, P = 0.02), larger adipocytes (P = 0.004), and positive surgical margins at the time of surgery (P = 0.04). WATi correlated with higher circulating levels of insulin, triglycerides, and leptin/adiponectin ratio, and lower high density lipoprotein cholesterol, compared to pts without WATi (P’s &lt; 0.05). Conclusions: Periprostatic WATi is common in men with PC. It is associated with high grade PC and alterations in systemic factors that contribute to PC development and progression. Periprostatic WATi may represent a therapeutic target for improving PC risk and outcomes.

Abstract PD5-05: Metabolic obesity, adipose inflammation and aromatase: Potential drivers of breast cancer risk in women with normal body mass index

Abstract Background: Elevated body mass index (BMI) is associated with increased risk of postmenopausal breast cancer, which may be partly attributable to an inflammation-aromatase axis. Most individuals with elevated BMI harbor white adipose tissue inflammation (WATi), defined by the presence of crown-like structures in the breast (CLS-B). CLS-B are composed of a dead/dying adipocyte surrounded by CD68+ macrophages. This inflammation is associated with activation of NF-κB and elevated expression of aromatase, which could contribute to tumor development. Additionally, WATi correlates with several circulating changes, including hyperinsulinemia, which increase breast cancer risk. Although breast WATi correlates with rising BMI, it is also present in some normal BMI individuals. Beyond inherited germline syndromes, the etiology of breast cancer in individuals with normal BMI is not well understood. Here we examined the impact of breast WATi on breast aromatase expression and circulating factors in women with normal BMI. Methods: Non-tumorous breast tissue and fasting blood were collected from 72 women with BMI &amp;lt; 25 kg/m2 undergoing mastectomy at MSKCC. Breast inflammation was detected by the presence of CLS-B using CD68 immunohistochemistry. The primary objective was to determine if breast WATi in normal BMI individuals correlates with elevated aromatase levels in the breast, measured by qPCR, western blotting, immunofluorescence and enzyme activity. Secondary objectives included assessment of breast adipocyte size and circulating metabolic and inflammatory factors. Results: Breast inflammation was present in 39% of women. Median BMI was 23.0 (range 18.4 to 24.9) in women with breast WATi versus 21.8 (range 17.3 to 24.6) in those without inflammation (P=0.04). Aromatase mRNA expression was positively correlated with WATi (CLS-B/cm2; P=0.002). Those with severe WATi had highest aromatase mRNA levels, compared to those with no or mild WATi (P=0.005). Aromatase protein, assessed by measuring adipose stromal cell-specific immunofluorescence or western blotting, and activity were also higher in CLS-B+ cases compared to CLS-B- (P&amp;lt;0.001). Breast WATi correlated with larger adipocytes (P=0.01) and higher circulating levels of C-reactive protein, leptin, insulin, and triglycerides (P&amp;lt;0.05). Insulin resistance, characterized by the homeostasis model (HOMA2-IR), correlated with breast WATi (P=0.004). Finally, leptin, a known inducer of aromatase and driver of cancer growth, correlated with higher breast aromatase levels (P=0.02) and larger adipocytes (P&amp;lt;0.01). Conclusions: A metabolically unhealthy state occurs in women with inflamed breast adipose despite having a normal BMI. This subclinical inflammatory state is characterized by elevated aromatase in the breast, insulin resistance, and dysplipidemia. The presence of enlarged adipocytes in the breasts of normal BMI women with inflammation suggests a state of hyperadiposity which could not be predicted based on BMI alone. These findings indicate that normal BMI metabolic obesity may be associated with increased cancer risk. Our results suggest that objective measurements of adiposity rather than BMI may help to identify individuals at increased risk for disease. Citation Format: Iyengar NM, Brown KA, Zhou XK, Subbaramaiah K, Giri DD, Gucalp A, Howe LR, Zahid H, Bhardwaj P, Wendel NK, Falcone DJ, Morrow M, Wang H, Williams S, Pollak M, Hudis CA, Dannenberg AJ. Metabolic obesity, adipose inflammation and aromatase: Potential drivers of breast cancer risk in women with normal body mass index [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr PD5-05.

Abstract 2767: Investigation of the relationship between crown-like structures and adipose tissue hormone levels among postmenopausal women with breast cancer

Abstract Introduction: Increased levels of circulating estrogens are associated with elevated risks of postmenopausal breast cancer. Previously, we showed that levels of sex steroid hormones in breast adipose tissue are positively correlated with circulating hormones. Further evaluation of hormone levels in breast tissues may help to explain the relationship between estrogens and breast cancer risk. Specifically, data suggest that focal breast lesions consisting of macrophages encircling dying adipocytes, referred to as “crown-like structures (CLS)”, are associated with increased aromatase activity, which could increase conversion of androgens to estrogens locally in the breast. Therefore, we assessed whether the presence of CLS affects the correlations between blood and breast adipose tissue hormone levels. Methods: Participants included 84 postmenopausal women with invasive breast cancer in the Polish Breast Cancer Study, a population based case-control study conducted from 2000 to 2003. We previously reported sex steroid hormone levels in cancer-free breast adipose tissue removed as part of breast excisions performed for cancer. Here, using immunohistochemical stains, we evaluated the number of CD68 positive macrophages and the presence of full CLS (i.e., complete adipocyte encirclement) in benign breast tissue sections prepared from surgical pathology specimens containing breast cancer. The relationship between the number of CD68 positive macrophages per unit area of fat and hormone levels was examined using Spearman rank correlations (rs). We assessed potential effect modification of the association between blood and tissue hormone levels by presence or absence of CLS by including an interaction term in linear regression models. Results: The number of CD68 positive macrophages per unit area of fat was not significantly correlated with levels of estrone, estradiol, androstenedione or testosterone measured in blood (rs = 0.001, -0.04, -0.07, and -0.09, respectively; p&amp;gt;0.05) or breast adipose tissue (rs = -0.15, 0.06, -0.06, and -0.08, respectively; p&amp;gt;0.05). Correlations between serum and adipose tissue hormones did not statistically significantly differ between women with CLS (n = 30) compared to those without CLS (n = 54). CLS were found more frequently in breast adipose tissue from overweight (i.e., women with a body mass index (BMI) between 25 and 29.9) (43%) and obese women (BMI &amp;gt;30) (30%) versus normal weight women, with (BMI between 18.5 and 24.9), (27%); (p = 0.04, χ2 test). The number of CD68 positive macrophages per unit area of fat was not significantly related to BMI (p = 0.31). Conclusion: Among postmenopausal women with invasive breast cancer, the presence of CLS was more common among overweight and obese women, but CLS did not influence the correlation between circulating and breast adipose tissue sex steroid hormone levels. Citation Format: Maeve Mullooly, Hannah P. Yang, Roni T. Falk, Sarah Nyante, Renata Cora, Ruth M. Pfeiffer, Derek C. Radisky, Daniel W. Visscher, Lynn C. Hartmann, Amy C. Degnim, Frank Z. Stanczyk, Jonine F. Figueroa, Montserrat Garcia-Closas, Jolanta Lissowska, Melissa T. Troester, Louise A. Brinton, Mark E. Sherman, Gretchen L. Gierach. Investigation of the relationship between crown-like structures and adipose tissue hormone levels among postmenopausal women with breast cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2767. doi:10.1158/1538-7445.AM2015-2767

Pilot study evaluating presence of crown-like structures in high grade endometrial carcinoma.

e16504 Background: Obesity is associated with an increased incidence of Type I & II endometrial cancer (EC), decreased recurrence-free interval, and significantly increased mortality rate in EC. Obesity-induced pro-inflammatory state, hyperglycemia, elevated IGF-1, and subsequent mTOR pathway activation are all implicated in endometrial carcinogenesis. Crown-like structures (CLS), consisting of dead or dying adipocytes surrounded by macrophages, are a marker of obesity-associated white adipose tissue (WAT) inflammation (WATi). Their presence in EC has never been examined. Methods: From 2009-2012, women with stage III or IV Grade 3 endometrioid or serous EC who underwent primary surgery at MSKCC and consented to an institutional tissue-banking protocol were identified. WAT was examined in perinodal tissue and omentum and subjected to immunostain with the macrophage marker, CD68, in order to detect CLS by light microscopy. WAT area was measured with NIH Image J in order to provide a standardized measurement...

Abstract P4-04-12: Both metabolic syndrome and statin use are more common in women with breast inflammation

Abstract Background: Metabolic syndrome is associated with increased breast cancer (BC) risk. After menopause, obesity is associated with increased risk of hormone receptor (HR)-positive breast cancer. We demonstrated an obesity → inflammation → aromatase axis in breast white adipose tissue (WAT) where inflammation is defined by the presence of crown-like structures (CLS) consisting of a dead or dying adipocyte encircled by macrophages. CLS in the breast (CLS-B) are associated with elevated body mass index (BMI), postmenopausal status, increased adipocyte size, and increased tissue levels of proinflammatory mediators and aromatase. As obesity is a component of the metabolic syndrome and is associated with inflammation, we compared levels of relevant circulating factors in women with and without breast WAT inflammation. Methods: We prospectively collected paired WAT and fasting serum and plasma from women undergoing mastectomy at MSKCC. WAT inflammation was detected by CD68 immunohistochemistry to identify CLS-B by light microscopy. Plasma levels of glucose, insulin, hsCRP, leptin, adiponectin, and IL-6 were measured by ELISA. Serum levels of total cholesterol, triglycerides (TG), HDL and LDL cholesterol were determined. Insulin resistance (IR) was assessed using fasting plasma glucose and insulin levels via the updated Homeostasis Model Assessment (HOMA2-IR). Associations between CLS-B and clinicopathologic features, including medication usage, were analyzed by logistic regression and Fisher’s exact test. Differences in serum/plasma endpoints between subjects with and without CLS-B were evaluated using Student t-test and nonparametric Wilcoxon rank-sum test. Results: From 11/2011 – 3/2013 we accrued 100 patients (pts); median age 47 years (range 27 – 70). Overall, CLS-B were found in 52/100 (52%) pts: 18/19 (95%) obese pts, 17/33 (52%) overweight pts, and 17/48 (35%) normal BMI pts. A clinical diagnosis of dyslipidemia was present in 14/52 (27%) pts with CLS-B and 1/48 (2%) pts without CLS-B (P&amp;lt;0.001). CLS-B were found in 10/11 (91%) statin users, but in only 42/89 (47%) non-users (P=0.008). Fasting glucose, insulin, LDL, TG, leptin, hsCRP, and IL-6 levels were higher in pts with CLS-B (Table). HOMA2-IR was greater in pts with CLS-B (mean 0.63 ±0.34) versus those without CLS-B (mean 0.46 ±0.23; P=0.006). Fasting level, mean (SD)No CLS-B, N=48CLS-B +, N=52PGlucose, mg/dL73.2 (8.0)84.3 (37.6)0.04Insulin, mU/L4.3 (2.1)5.6 (2.9)0.01LDL cholesterol, mg/dL105.9 (31.0)119.4 (32.4)0.04HDL cholesterol, mg/dL71.9 (15.8)62.1 (16.1)0.003Triglycerides, mg/dL69.2 (26.3)104.9 (50.6)&amp;lt;0.001Leptin, pg/mL12.0 (10.1)22.6 (19.7)&amp;lt;0.001Adiponectin, ng/mL13.7 (5.2)10.4 (5.4)0.002hsCRP, ng/mL1.0 (1.4)2.3 (2.7)0.003 Conclusions: Breast WAT inflammation, which we have previously linked to increased aromatase activity, is associated with biochemical changes that occur in the metabolic syndrome, a risk factor for BC. Statin use is more common in patients with breast WAT inflammation and metabolic syndrome. Clinically, statin use may be a surrogate identifier of a population that is at increased baseline risk of BC. These findings may account for the variability in results of prior studies examining statin use and breast cancer risk due to elevated risk in users compared to non-users. Citation Format: Neil M Iyengar, Ayca Gucalp, Xi K Zhou, Louise R Howe, Patrick G Morris, Dilip Giri, Kotha Subbaramaiah, Priya Bhardwaj, Samuel S Park, Michael Pollak, Monica Morrow, Clifford A Hudis, Andrew J Dannenberg. Both metabolic syndrome and statin use are more common in women with breast inflammation [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-04-12.