Genetic variation in one-carbon metabolism in relation to genome-wide DNA methylation in breast tissue from heathy women.

Single nucleotide polymorphisms (SNPs) in one-carbon metabolism genes and lifestyle factors (alcohol drinking and breast folate) may be determinants of whole-genome methylation in the breast. DNA methylation profiling was performed using the Illumina Infinium HumanMethylation450 BeadChip in 81 normal breast tissues from women undergoing reduction mammoplasty and no history of cancer. ANCOVA, adjusting for age, race and BMI, was used to identify differentially-methylated (DM) CpGs. Gene expression, by the Affymetrix GeneChip Human Transcriptome Array 2.0, was correlated with DM. Biological networks of DM genes were assigned using Ingenuity Pathway Analysis. Fifty-seven CpG sites were DM in association with eight SNPs in FTHFD, MTHFD1, MTHFR, MTR, MTRR, and TYMS (P <5.0 x 10-5); 56% of the DM CpGs were associated with FTHFD SNPs, including DM within FTHFD. Gene expression was negatively correlated with FTHFD methylation (r=-0.25, P=0.017). Four DM CpGs identified by SNPs in MTRR, MTHFR, and FTHFD were significantly associated with alcohol consumption and/or breast folate. The top biological network of DM CpGs was associated with Energy Production, Molecular Transportation, and Nucleic Acid Metabolism. This is the first comprehensive study of the association between SNPs in one-carbon metabolism genes and genome-wide DNA methylation in normal breast tissues. These SNPs, especially FTHFD, as well as alcohol intake and folate exposure, appear to affect DM in breast tissues of healthy women. The finding that SNPs in FTHFD and MTR are associated with their own methylation is novel and highlights a role for these SNPs as cis-methylation quantitative trait loci.

Relationships among folate, alcohol consumption, gene variants in one-carbon metabolism and p16INK4a methylation and expression in healthy breast tissues.

p16(INK4a) is a tumor suppressor gene, frequently hypermethylated in breast cancer; this epigenetic silencing of p16(INK4a) occurs early in carcinogenesis. The risk factors and functional consequences of p16(INK4a) methylation are unknown. Alcohol consumption, a breast cancer risk factor, impedes folate metabolism and may thereby alter gene methylation since folate plays a pivotal role in DNA methylation. In a cross-sectional study of 138 women with no history of breast cancer who underwent reduction mammoplasty, we studied breast cancer risk factors, plasma and breast folate concentrations, variation in one-carbon metabolism genes, p16(INK4a) promoter methylation and P16 protein expression. Logistic regression was used to estimate multivariable-adjusted odds ratios (OR) and 95% confidence intervals (CI). p16(INK4a) methylation was negatively correlated with P16 expression (r = -0.28; P = 0.002). Alcohol consumption was associated with lower breast folate (P = 0.03), higher p16(INK4a) promoter methylation (P = 0.007) and less P16 expression (P = 0.002). Higher breast folate concentrations were associated with lower p16(INK4a) promoter methylation (P = 0.06). Genetic variation in MTRR (rs1801394) and MTHFD1 (rs1950902) was associated with higher p16 (INK4a) promoter methylation (OR = 2.66, 95% CI: 1.11-6.42 and OR = 2.72, 95% CI: 1.12-6.66, respectively), whereas variation in TYMS (rs502396) was associated with less P16 protein expression (OR = 0.22, 95% CI: 0.05-0.99). Given that this is the first study to indicate that alcohol consumption, breast folate and variation in one-carbon metabolism genes are associated with p16(INK4a) promoter methylation and P16 protein expression in healthy tissues; these findings require replication.

A prospective study on lipoaugmentation of the breast.

BACKGROUND: The current standard for breast augmentation involves placement of an implant. As an alternative, surgeons have been exploring breast augmentation with autologous tissue in the form of injectable fat. OBJECTIVES: The authors explore the efficacy and safety of lipoaugmentation of the breast, with specific interest in volume changes, fat retention, overall aesthetic improvement, and patient satisfaction. METHODS: Direct measurements, 2- and 3-dimensional images, mammograms, and magnetic resonance imaging (MRI) were obtained preoperatively from 10 consecutive patients undergoing augmentation mammaplasty with autologous fat transfer. These measurements were repeated 1 year postoperatively. Postoperative photo imaging was conducted at 3-month intervals for 1 year. Efficacy was evaluated by determining the volume of fat retention 1 year after the procedure with 3-dimensional imaging, standard breast MRI volume measurements, and subjective aesthetic comparisons. RESULTS: The average amount of fat injected was 236 cc (90-324; SD, 69.8) in the right breast and 250 cc (90-300; SD, 65.1) in the left. The mean volume change based on 3-dimensional imaging was 85.1 cc (36% retention) for the right breast and 98.1 cc (39.2% retention) for the left. The mean volume change based on MRI measurements was 30.0 cc (39.8% change) on the right and 29.3 cc (38.1% change) on the left. Blinded observers found substantial improvement in 1 patient (10%), moderate improvement in 5 patients (50%), and minimal to no improvement in 4 (40%). Overall patient satisfaction was high, as measured by the abbreviated BREAST-Q. Radiologic abnormalities and artifacts were common and required additional imaging. CONCLUSIONS: Objective breast enlargement in this study was modest but yielded disproportionately high subjective patient satisfaction reports. LEVEL OF EVIDENCE: 2.

Reduction mammaplasty and mastopexy in previously irradiated breasts.

BACKGROUND: Little data exist on the safety of elective breast surgery following breast conservation therapy. OBJECTIVES: The authors evaluate their experience performing reduction mammaplasties and mastopexies in previously irradiated breasts. METHODS: A retrospective chart review was conducted of all reduction mammaplasties and mastopexies in previously irradiated breasts performed by the 2 senior authors (MYN and SLS) from 1995 to 2012 (n = 18). Patient records were analyzed for demographics, perioperative and postoperative details, and complications. RESULTS: During the study period, 12 reduction mammaplasties and 6 mastopexies were performed on breasts previously treated with breast conservation therapy. The average study participant age was 49.5 years, and average body mass index was 29. Average preoperative bra cup size was D/DD. One patient was a former smoker; 17 were nonsmokers. Eleven of the 18 patients had been previously treated with chemotherapy; average time between radiation therapy and surgery was 2.5 years. Average specimen weight in the 12 reduction mammaplasties was 623 g. Twenty-two percent (n = 4) of patients experienced a total of 5 minor complications; none required a return to the operating room. One patient (6%) experienced a major complication resulting in approximately 50% loss of breast tissue, which was reconstructed with a latissimus dorsi musculocutaneous flap. Seventeen patients (94%) had successful outcomes with preoperative goals achieved. CONCLUSIONS: In the previously irradiated breast, reduction mammaplasty and mastopexy should be approached with extra caution, but the surgeries may still be safely and successfully performed. LEVEL OF EVIDENCE: 4.

Plasma IGF-1 and IGFBP-3 may be imprecise surrogates for breast concentrations: an analysis of healthy women.

We investigated insulin-like growth factor (IGF)-1 and IGF binding protein (IGFBP)-3 concentrations in histologically normal breast tissues and assessed their association with plasma concentrations, and breast cancer risk factors. IGF-1 and IGFBP-3 were assessed in plasma and breast tissues of 90 women with no history of any cancer and undergoing reduction mammoplasty. Pearson correlations and ANOVAs were used to describe plasma-breast associations and biomarker differences by breast cancer risk factors, respectively. Multivariable regression models were used to determine associations between risk factors, and breast IGF-1 and IGFBP-3. The mean age of the study sample was 37.3 years, 58 % were white, and generally these women were obese (mean BMI = 30.8 kg/m(2)). We observed no plasma-breast correlation for IGF-1, IGFBP-3, or IGF-1/IGFBP-3 (r = -0.08, r = 0.14, and r = 0.03, respectively; p-values >0.05). Through age- and BMI-adjusted analysis, BMI and years of oral contraceptive (OC) use were inversely associated with breast IGF-1 (p-values = 0.02 and 0.003, respectively) and age was associated with breast IGFBP-3 (p = 0.01), while breast IGF-1/IGFBP-3 was higher in blacks than whites (1.08 vs. 0.68, p = 0.04) and associated with age and BMI (p-values = 0.03 and 0.002, respectively). In multivariable-adjusted models, some breast cancer risk factors studied herein explained 24, 10, and 15 % of the variation in breast IGF-1, IGFBP-3, and IGF-1/IGFBP-3, respectively. While reasons for the lack of plasma-breast hormone correlations in these cancer-free women are unknown, several factors were shown to be associated with breast concentrations. The lack of correlation between blood and tissue IGF-1 and IGFBP-3 suggests that studies of breast cancer risk assessing blood IGF-1 and IGFBP-3 may have important limitations in understanding their role in breast carcinogenesis.

Long-term outcomes of failed prosthetic breast reconstruction.

INTRODUCTION: For patients undergoing prosthetic breast reconstruction whose device was removed because of infection or exposure, there is no published information examining long-term outcomes. Despite initial failure, many patients want to pursue breast reconstruction. METHODS: A retrospective review was performed of immediate prosthetic breast reconstruction failures during a 5-year period. RESULTS: A total of 26 patients (29 breasts) were identified who failed primary prosthetic 2-stage reconstruction. On average, the device was removed 262 days from the initial expander placement. Of these breasts, 19 were removed for infection, 7 for exposure, and 3 for a combination of both. Of these 29 failures, secondary reconstruction was attempted in 20 (69%). Among secondary reconstructions, 13 were with a device and 7 with autologous tissue. Of the 13 (92.3%) secondary device reconstructions, 12 were successful.Those in whom secondary reconstruction was attempted were younger at initial reconstruction (48 vs 57 years; P = 0.038), had lower body mass indexes (BMIs) (26.0 vs 29.4; P = 0.27), and smaller breasts (650 vs 979 g; P = 0.23) than those who did not attempt secondary reconstruction.Of the nipple-sparing mastectomy patients, 100% underwent secondary reconstruction, whereas only 61% of skin-sparing mastectomy patients underwent secondary reconstruction (P = 0.14).For patients undergoing secondary reconstruction, those receiving a second device reconstruction had lower BMIs (24.7 vs 28.5; P = 0.18) and smaller breasts (489 vs 946 g; P = 0.08) than those with autologous reconstructions; 15% of secondary implant reconstructions underwent irradiation versus 43% of the secondary autologous reconstructions (P = 0.29). CONCLUSIONS: After failed prosthetic breast reconstruction, a second attempt with an implant in properly selected patients has a high success rate (92.3%).

Effect of incision choice on outcomes in primary breast augmentation.

BACKGROUND: Capsular contracture (CC) is the most common complication following primary breast augmentation and one of the most common causes of reoperation. Various studies have suggested certain risk factors, including incision choice. OBJECTIVES: The authors investigate a possible association between the three most common breast augmentation incisions (inframammary, periareolar, and transaxillary) and CC. METHODS: The authors conducted a retrospective chart review of 197 primary breast augmentation patients treated between 2003 and 2009. Significant CC was determined to have occurred if the patient required reoperation for her CC. Patients were excluded if they underwent an augmentation/mastopexy, had previously undergone breast surgery, or received shaped silicone gel implants. CC rates were analyzed on a per-patient basis with Fisher's exact test and on a per-breast basis with the Rao-Scott chi-squared test. RESULTS: One hundred eighty-three patients (336 augmented breasts) were included. Average patient age was 36.5 years. Mean follow-up was 392.6 days. Surgical complications included six breasts with CC (1.8%), three with hematoma (0.9%), and one with an infection (0.3%). Transaxillary incisions produced the highest incidence of contracture (6.4%), followed by periareolar (2.4%) and inframammary (0.5%). There was a statistically-significant difference in the incidence of CC among the three incision sites (P=.03). The increased rate seen with transaxillary incisions versus inframammary incisions was also statistically-significant. No significant association between implant fill material and contracture was found (P=.27). CONCLUSIONS: The risk of CC is significantly higher with transaxillary incisions than with periareolar or inframammary incisions. LEVEL OF EVIDENCE: 4.

Acellular dermal matrix for secondary procedures following prosthetic breast reconstruction.

Acellular dermal matrices (ADM) have generated interest for their possible applications in secondary revisions following prosthetic breast reconstruction. These materials can be effective in a variety of situations, including implant displacement, synmastia, capsular contracture, incisional support, and pocket conversion. ADM can also be placed in the setting of delayed breast reconstruction and to augment nipple projection. These biomaterials have demonstrated feasibility and success for many complex deformities. However, there is an associated learning curve that includes an understanding of proper technique and patient selection. The authors review their cumulative experience between 2004 and 2010 with ADM for the correction of secondary deformities following prosthetic breast reconstruction, focusing on the indications for repair, traditional management, and management with ADM.

Breast implants: saline or silicone?

The United States has seen significant shifts in the breast implant market over the past five decades. From the moratorium on silicone gel breast implants in 1992 to their approval in 2006, there have been many developments in their manufacturing and usage. Meanwhile, saline breast implants have remained somewhat unchanged, still offering a few distinct advantages but none of the technological innovation of the silicone gel models. In this article, the authors review the current state of silicone gel and saline implants, as well as the advantages and disadvantages of each. Much of the current data on complications of gel and saline implants are examined, as well as some practical implications associated with the use of each implant type.

Outcome assessment of breast distortion following submuscular breast augmentation.

BACKGROUND: Animation deformity or breast distortion during pectoralis muscle contraction following subpectoral breast augmentation is a known entity, but its prevalence and significance remain unclear. The purpose of this study was to identify the incidence and severity of animation deformity as well as its effect on patient satisfaction and interference with certain activities. METHODS: All procedures were performed by the senior author using a variation of a previously described dual-plane technique. The first part of this study was an evaluation of breast distortion by a group of independent observers in a series of 40 consecutive patients who underwent primary subpectoral breast augmentation. The second part of the study was a questionnaire sent to 195 consecutive patients asking about overall satisfaction, degree of animation deformity, and whether there was interference with any activities. RESULTS: Of the 40 patients' photographs that were evaluated, 9 (22.5%) had no distortion, 25 (62.5%) had minimal distortion, 4 (10%) had moderate distortion, and 2 (5%) had severe distortion. Of the 195 questionnaires, there were 69 responses, a 35% response rate. Fifty-six (82%) described mild to no distortion, 7 (10%) were moderate, and 5 (7%) were severe. According to the survey, the most common activities that were problematic were lifting weights and exercising (24 and 19%, respectively). Only one (1%) patient stated that she would not recommend subpectoral positioning. CONCLUSION: Although animation deformities do exist, nearly all patients in this study would still choose subpectoral positioning. Patients who may be better candidates for subglandular placement are those for whom exercise is central to their daily living. As a result of this study, surgeons and patients should have more accurate and reliable information regarding the significance of animation deformity after subpectoral breast augmentation.

Considerations of previous augmentation in subsequent breast reconstruction.

BACKGROUND: The number of patients receiving augmentation mammaplasty is rapidly growing. Breast cancer will develop in a significant number of these women. OBJECTIVE: The authors report on a series (expanding the size of a previous study by the senior author, S.L.S.), encompassing one surgeon's experience performing breast reconstruction in women who had undergone previous augmentation. They compare the stage and detection method in this series with other published studies and also review their experience with regard to the benefit of subpectoral versus subglandular implant placement. METHODS: A retrospective review of the senior author's reconstructive practice was performed, including the period from July 1983 to July 2007. Thirty-two consecutive women were identified who had previously received augmentation mammaplasty, were subsequently diagnosed with cancer, and then underwent breast reconstruction. Types of reconstructive procedures and outcomes were evaluated. A statistical analysis of the results was performed with a standard 2-tailed t test and chi(2) analysis. RESULTS: The occurrence of breast cancer diagnosis after augmentation ranged from 1 to 25 years (mean, 15 years). No implants were ruptured at the time of mastectomy. Of the 16 patients with previous subpectoral augmentation, cancer was detected mammographically in 12 (75%). Of the 16 patients with previous subglandular augmentation, cancer was detected mammographically in 7 (44%). This difference was not statistically significant, but that may be related to the insufficient population size (P = .15). Twenty-two (69%) of the patients underwent a purely prosthetic reconstruction. Flaps were used in the other 10 (31%), including 5 (16%) latissimus dorsi flaps and 5 (16%) transverse rectus abdominis flaps. Nine (90%) of those 10 flaps were used in patients undergoing radiation therapy. There were 9 stage 0 (28.1%), 9 stage I (28.1%), 12 stage II (37.5%), and 2 stage III (6.3%) patients. Among the subglandular group, there were 5 stage 0 (31.3%), 2 stage I (12.5%), 7 stage II (43.8%), and 2 stage III (12.5%) patients. Among the subpectoral group, there were 4 stage 0 (25.0%), 7 stage I (43.8%), 5 stage II (31.3%), and no stage III or IV patients. There was no significant difference in the axillary status between the 2 implant location groups. Eleven of the total 32 (34%) patients were treated with radiation therapy. Of these patients, 2 received breast conservation therapy and the other 9 underwent mastectomy. Of the 11 augmented breasts that received radiation therapy, 9 had flaps used in their reconstruction, including 5 latissimus dorsi and 4 trans-rectus abdominis muscle flaps. The average length of follow-up for the entire group was 26.4 months (range, 1-109 months). CONCLUSIONS: A history of breast augmentation was demonstrated to have consequences for future management of cancer in the areas of detection, cancer management, and reconstruction options. In addition, it was shown that the site of implantation may have an effect on the effectiveness of breast imaging.

Breast Cancer following Augmentation Mammaplasty: A Case-Control Study.

BACKGROUND: The purpose of this study was to determine whether augmentation mammaplasty, implant type, and implant location affect breast cancer detection, stage, and treatment. METHODS: An institutional case-control study was performed of patients with prior breast augmentation undergoing breast cancer treatment from 2000 to 2013. Controls were propensity matched and randomized, and data were retrospectively reviewed. RESULTS: Forty-eight cases and 302 controls were analyzed. Palpable lesions were detected at a smaller size in augmentation patients (1.6 cm versus 2.3 cm; p < 0.001). Fewer lesions in augmented patients were detected by screening mammography (77.8 percent of cases versus 90.7 percent of controls; p = 0.010). Patients with implants were more likely to undergo an excisional biopsy for diagnosis (20.5 percent versus 4.4 percent; p < 0.001), rather than image-guided core needle biopsy (77.3 percent versus 95.3 percent; p < 0.001). Earlier staging in augmented patients approached but did not reach statistical significance (p = 0.073). Augmented patients had higher mastectomy rates (74.5 percent versus 57.0 percent) and lower rates of breast-conservation therapy (25.5 percent versus 43 percent; p = 0.023). Neither implant fill type nor anatomic location affected method of diagnosis, stage, or treatment. CONCLUSIONS: Palpable detection of breast cancer is more likely at a smaller size in augmented patients, yet it is less likely on screening mammography than in controls. Augmentation breast cancer patients have a comparable disease stage and are more likely to undergo mastectomy rather than lumpectomy. Both silicone and saline implants, whether placed submuscularly or subglandularly, have comparable effects on breast imaging, biopsy modality, and surgical intervention. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, III.

Implant-based reconstruction.

Prosthetic reconstruction of the breast, either with standard or adjustable implants or as a staged procedure with tissue expanders followed by implants, has evolved into a reliable method for breast reconstruction. Advances in implant technology and biomaterials allow for low complication rates, good aesthetic outcomes, and consistent results. Potential advantages over other reconstructive techniques include the relative simplicity of the procedures, the use of adjacent tissue without significant donor site morbidity, reduced operative time, and quicker patient recovery. Although some may still consider autologous tissue reconstruction as the gold standard, today's prosthetic reconstructions play an equally important role in the reconstruction of the breast for many women.

Allergan's silicone gel breast implants.

In the USA, women seeking breast implants for augmentation, revision or reconstruction can choose between saline-filled devices and round, silicone gel-filled devices. Form-stable, highly cohesive silicone gel-filled breast implants are marketed in other countries and are currently under review by the US FDA. Allergan has conducted clinical studies to investigate the safety and effectiveness of its round and anatomical (Style 410) devices for US marketing approval. The most frequently reported complications were reoperation, implant removal with replacement, implant malposition and capsular contracture. The FDA approved the round devices in 2006. The weight of the scientific literature suggests that silicone gel-filled breast implants do not increase a patient's risk of cancer, autoimmune disease, reproductive effects or suicide. As differently shaped, cohesive breast implants continue to be introduced, breast implant surgery will become more customized to the patient's biological conditions and desires.

Comparison of Different Acellular Dermal Matrices in Breast Reconstruction: The 50/50 Study.

BACKGROUND: Acellular dermal matrix has enjoyed extensive use in primary and secondary alloplastic breast aesthetic and reconstructive surgery. The objective of this study was to examine clinical outcomes between available acellular dermal matrix products: DermACELL (LifeNet Health, Virginia Beach, Va.) and AlloDerm Ready To Use (LifeCell Corp., Branchburg, N.J.). METHODS: A retrospective chart review was performed on 58 consecutive patients (100 breasts) reconstructed with either DermACELL (n = 30 patients; 50 breasts) or AlloDerm Ready To Use (n = 28 patients; 50 breasts). The mastectomies were performed by three different breast surgeons. All reconstructions were performed by the same plastic surgeon (T.A.P.). Statistical analysis was performed by means of Fisher's exact test. RESULTS: Differences in the average age, body mass index, percentage having neoadjuvant/adjuvant chemotherapy or breast irradiation, and numbers of therapeutic and prophylactic mastectomies between the two groups were not statistically significant (p < 0.05). Complications in both cohorts of patients were clinically recorded for 90 days after immediate reconstruction. When comparing outcomes, patients in the DermACELL group had a significantly lower incidence of "red breast syndrome" (0 percent versus 26 percent; p = 0.0001) and fewer days before drain removal (15.8 days versus 20.6 days; p = 0.017). No significant differences were seen in terms of seroma, hematoma, delayed healing, infection, flap necrosis, or explantation. CONCLUSION: Patients reconstructed with DermACELL as compared with AlloDerm Ready To Use have significantly decreased number of days to drain removal and red breast syndrome and equivalent rates of other complications, including seroma, infection, flap necrosis, and explantation. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, III.

Landscape of genome-wide age-related DNA methylation in breast tissue.

Despite known age-related DNA methylation (aDNAm) changes in breast tumors, little is known about aDNAm in normal breast tissues. Breast tissues from a cross-sectional study of 121 cancer-free women, were assayed for genome-wide DNA methylation. mRNA expression was assayed by microarray technology. Analysis of covariance was used to identify aDNAm's. Altered methylation was correlated with expression of the corresponding gene and with DNA methyltransferase protein DNMT3A, assayed by immunohistochemistry. Publically-available TCGA-BRCA data were used for replication. 1,214 aDNAm's were identified; 97% with increased methylation, and all on autosomes. Sites with increased methylation were predominantly in CpG lslands and non-enhancers. aDNAm's with decreased methylation were generally located in intergenic regions, non-CpG Islands, and enhancers. Of the aDNAm's identified, 650 are known to be involved in cancer, including ESR1 and beta-estradiol responsive genes. Expression of DNMT3A was positively associated with age. Two aDNAm's showed borderline significant associations with DNMT3A expression; KRR1 (OR 6.57, 95% CI: 2.51-17.23) and DHRS12 (OR 6.08, 95% CI: 2.33-15.86). A subset of aDNAm's co-localized within vulnerable regions for somatic mutations in cancers including breast cancer. Expression of C19orf48 was inversely and significantly correlated with its methylation level. In the TCGA dataset, 84% and 64% of the previously identified aDNAm's were correlated with age in both normal-adjacent and tumor breast tissues, with differential associations by histological subtype. Given the similarity of findings in the breast tissues of healthy women and breast tumors, aDNAm's may be one pathway for increased breast cancer risk with age.