Min Kyoon Kim

Predictive Significance of p53, Ki-67, and Bcl-2 Expression for Pathologic Complete Response after Neoadjuvant Chemotherapy for Triple-Negative Breast Cancer

Purpose Patients with triple-negative breast cancer (TNBC) with pathologic complete response (pCR) to neoadjuvant chemotherapy (NAC) have superior survival outcomes compared to those with residual disease after NAC. This study investigated the value of three biomarkers, p53, Ki-67, and Bcl-2 for predicting pCR in NAC-treated patients with TNBC. Methods Between 2003 and 2012, 198 patients with pathologically confirmed primary TNBC were treated with two different taxane-based chemotherapeutic regimens prior to surgery. Before NAC, expression of p53 (cutoff 25%), Ki-67 (cutoff 10%), and Bcl-2 (cutoff 10%) was assessed immunohistochemically in core biopsy specimens. The incidence of pCR was correlated with the expression of these biomarkers. Results Overall, pCR occurred in 37 of the 198 patients (18.7%). A significant association was observed between the pCR rate and overexpression of the p53 and Ki-67 biomarkers. Multivariate analysis showed that only p53 expression was independently associated with pCR to NAC (odds ratio, 3.961; p=0.003). The sensitivity, specificity, positive predictive value, and negative predictive value of p53 expression for predicting pCR were 77.8%, 50.3%, 26.2%, and 90.9%, respectively. The pCR rate was the lowest (5.2%) in patients with low expression of both p53 and Ki-67, and it was the highest (25.8%) when both biomarkers showed high expression. Conclusion Expression of p53 was significantly associated with pCR after NAC in patients with TNBC, suggesting that this biomarker might be particularly valuable in identifying TNBC patients prone to have residual disease after NAC.

Prognostic effect of preoperative serum estradiol level in postmenopausal breast cancer

BackgroundThe prognostic role of serum estrogen level in breast cancer patients is unclear. We investigated the prognostic importance of preoperative serum estradiol (E2) level in postmenopausal women according to their estrogen receptor (ER) status.MethodsThe medical records of 313 postmenopausal breast cancer patients who underwent surgery between 2006 and 2008 at a single institution were retrospectively evaluated. Patients who received neoadjuvant chemotherapy, synchronous bilateral breast cancer, or those with metastasis at diagnosis were excluded. Serum E2 and follicular stimulating hormone (FSH) levels were measured by radioimmunoassay and immunoradiometric assay, respectively, within 3 months prior to surgery. After a median follow-up of 52.0 months (11–77 months), 21 women were found to have metastatic disease.ResultsThe overall, median E2 level was 13.0 pg/ml, and was slightly higher in ER-positive than ER-negative (p=0.69). The mean serum E2 level was significantly higher in patients with metastasis (17.41±8.34 pg/ml) than in those without metastasis (13.54±7.58 pg/ml) (p=0.02). Kaplan-Meier analysis using a cut-off of 13 pg/ml showed that, ER negative (p=0.02) but not ER positive (p>0.05) patients with higher E2 level showed significantly poorer metastasis-free survival. Multivariate analysis showed that, the high E2 level of ER negative tumors was an independent negative prognostic factor for metastasis- free survival (HR, 3.32; 95% CI, 1.05 to 10.51; p=0.04).ConclusionsHigher preoperative serum E2 level had a negative prognostic effect in postmenopausal women with breast cancer, especially in the ER-negative subgroup.

In Vivo Tumor Growth Rate Measured by US in Preoperative Period and Long Term Disease Outcome in Breast Cancer Patients

Objective The aim of our study was to evaluate the effect of tumor growth rate, calculated from tumor size measurements by US, on breast cancer patients’ outcome. Patients and Methods Breast cancer patients who received at least two serial breast ultrasonographies (US) in our institution during preoperative period and were surgically treated between 2002 and 2010 were reviewed. Tumor growth rate was determined by specific growth rate (SGR) using the two time point tumor sizes by US. Results A total of 957 patients were analyzed. The median duration between initial and second US was 28 days (range, 8–140). The median initial tumor size was 1.7cm (range, 0.4–7.0) and median second size was 1.9cm (range, 0.3–7.2). 523(54.6%) cases had increase in size. The median SGR(x10-2) was 0.59 (range, -11.90~31.49) and mean tumor doubling time was 14.51 days. Tumor growth rate was higher when initial tumor size was smaller. Lymphovascular invasion, axillary lymph node metastasis, and higher histologic grade were significantly associated with higher SGR. SGR was significantly associated with disease-free survival (DFS) in a univariate analysis (p = 0.04), but not in a multivariate Cox analysis (p>0.05). High SGR was significantly associated with worse DFS in a subgroup of initial tumor size >2cm (p = 0.018), but not in those with tumor size <2cm (p>0.05). Conclusion Our results showed that tumor growth rate measured by US in a relatively short time interval was associated with other worse prognostic factors and DFS, but it was not an independent prognostic factor in breast cancer patients.

Grade of ductal carcinoma in situ accompanying infiltrating ductal carcinoma as an independent prognostic factor.

BACKGROUND Several studies about the relationship between IDC and DCIS have been reported, but no consensus has been reached regarding clinical characteristics and prognostic value. PATIENTS AND METHODS We reviewed the medical records of patients who underwent surgery for IDC between 2006 and 2008. DCIS adjacent to IDC was pathologically classified as either high-grade DCIS or non-high-grade DCIS. RESULTS Among 1751 IDC patients within the study period, 1384 patients (79.0%) had concomitant DCIS. There was no survival difference between patients with pure IDC and those with IDC and concomitant DCIS. However, patients with high-grade DCIS had worse survival than did patients with non-high-grade DCIS or pure IDC (5-year recurrence-free survival rates for IDC with non-high-grade DCIS, pure IDC without DCIS, and IDC with high-grade DCIS were 97%, 93%, and 86%, respectively; P = .001). This tendency was maintained regardless of estrogen receptor status or histologic grade of IDC. In a Cox regression model, patients with IDC and accompanying high-grade DCIS had a 2.5-fold higher probability of local or distant relapse than did those with IDC and low-grade DCIS (hazard ratio, 2.51; 95% confidence interval, 1.12-5.64). CONCLUSIONS The prognosis of patients with invasive breast cancer differed according to the grade of concomitant adjacent DCIS. Accordingly, the grade of adjacent DCIS should be considered as a prognostic factor in the clinical management of patients with breast cancer. However, in our study, the follow-up periods were short to confirm prognostic effect. Further studies are needed.

Nomogram for Predicting Breast Conservation after Neoadjuvant Chemotherapy

Purpose The ability to accurately predict the likelihood of achieving breast conservation surgery (BCS) after neoadjuvant chemotherapy (NCT) is important in deciding whether NCT or surgery should be the first-line treatment in patients with operable breast cancers. Materials and Methods We reviewed the data of 513 women, who had stage II or III breast cancer and received NCT and surgery from a single institution. The ability of various clinicopathologic factors to predict the achievement of BCS and tumor size reduction to ≤ 3 cm was assessed. Nomograms were built and validated in an independent cohort. Results BCS was performed in 50.1% of patients, with 42.2% of tumors reduced to ≤ 3 cm after NCT. A multivariate logistic regression analysis showed that smaller initial tumor size, longer distance between the lesion and the nipple, absence of suspicious calcifications on mammography, and a single tumor were associated with BCS rather than mastectomy (p < 0.05). Negative estrogen receptor, smaller initial tumor size, higher Ki-67 level, and absence of in situ component were associated with residual tumor size ≤ 3 cm (p < 0.05). Two nomograms were developed using these factors. The areas under the receiver operating characteristic curves for nomograms predicting BCS and residual tumor ≤ 3 cm were 0.800 and 0.777, respectively. The calibration plots showed good agreement between the predicted and actual probabilities. Conclusion We have established a model with novel factors that predicts BCS and residual tumor size after NCT. This model can help in making treatment decisions for patients who are candidates for NCT.

Abstract P2-03-10: Comprehensive somatic SNV and CNV profiling for triple-negative breast cancer patients by targeted next-generation sequencing

Introduction: Triple-negative breast cancer is a subtype of breast cancer lacking expression of estrogen receptor, progesterone receptor and HER2/neu protein markers. Despite having the worse prognosis, currently there is no molecular target for this subtype. To discover the clues for therapeutic targets of TNBC, we examined somatic Single Nucleotide Variation (SNV) and Copy Number Variation (CNV) profiling of TNBC patients using targeted next-generation sequencing (NGS). Methods: A total of 414 breast cancer and normal breast samples were collected at Seoul National University Hospital (SNUH) from 1995 to 2013. By sample condition, 94 samples were frozen tissues (47 tumor-normal matched pair), and 320 samples were formalin-fixed and paraffin-embedded (FFPE) tissues (155 tumor-normal matched pair; Tumor 164, Normal 156). Genomic DNA was extracted from samples and target enrichment was done by using Agilent SureSelect Human Kinome Panel (612 genes including over 500 kinases). The paired-end libraries were constructed and sequenced on Illumina HiSeq 2000 instrument with average 250x depth coverage. Generated sequence reads were aligned to human genome hg19 with bwa algorithm, and somatic SNV and CNV were identified using VarScan2 algorithm. To confirm the existence of identified somatic SNV and CNV, we carried out SNP microarray experiment (Illumina Human Omni5 Exome microarray) for 46 pairs of frozen tissue samples. There were 18,720 SNP probes in SNP microarray covering target region of kinome panel. If the same SNV and CNV were detected in both NGS and microarray data, we considered them validated. Results: In 46 paired frozen samples, 610 somatic SNVs were detected. The most frequent and non-synonymous mutations were in PIK3CA (8.6%), BMPR1A (4.3%), FES (4.3%), TP53 (4.3%), ROCK1 (4.3%), PAK2 (4.3%), CDK18 (4.3%), OBSCN (4.3%), LRRK1 (4.3%), CDC6 (4.3%), PIKFYVE (4.3%), and FGFR3 (4.3%). Chromosomal aberration was detected in chromosome 1 (10 samples) and chromosome 8 (11 samples). 242 somatic CNV were found in 157 genes in 46 patients, but 99 gene amplifications showed only single occurrence. In other words, TNBC sample displayed highly heterogeneous CNV profile. However, we detected two frequently amplified genes; PKHD1L1 (13%) and NRBP2 (10.9%). Further analysis on NGS data of 155 pairs of FFPE samples is in process to validate the results. Conclusion : We investigated comprehensive somatic mutation profile in matched pair TNBC samples. Although these samples showed highly heterogeneous mutation profile, it’s possible to detect some interesting SNV and CNV with over 4% frequency. Further analysis may illuminate clinical meaning of those alterations. Citation Format: Taegyun Yun, Byung-Chul Lee, Jungsun Park, Dongyoon Park, Tae-Kyung Yoo, Min Kyoon Kim, Wonshik Han. Comprehensive somatic SNV and CNV profiling for triple-negative breast cancer patients by targeted next-generation sequencing [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 P2-03-10.

Abstract P2-03-18: Discovery of novel amplified genes in primary breast cancer with copy number and gene expression analysis of whole exome and transcriptome sequencing data

Introduction: Copy number alteration of genome is common in breast cancer and tend to have more driver role than single point mutations. Traditionally, genome-wide analysis of DNA copy number changes were done by array CGH or SNP array method. Here, we did DNA whole exome sequencing (WES) and RNA-seq using Next Generation Sequencing (NGS) technology to find common genes or chromosomal regions of which DNA copy was highly amplified and at the same time RNA expression was also upregulated. Materials and Method: RNA and DNA were extracted fromfresh frozen tissues of 93 breast cancer patients. WES and RNA-seq were done using NGS technology (Illumina HiSeq 2000). As a control, normal DNA from all matched patients were also sequenced. GATK was used to gain mean depth and coverage data for targeted regions.CNVs were calculated with ExomeCNV, a statistical method to detect somatic CNVs using depth-of-coverage information from mapped short sequence reads.To estimate expression levels, the relative transcript abundances were measured in FPKM using Cufflinks. Results and Discussion: DNA of 1,737 genes were highly amplified (log R>1.0) in two or more samples. The two most commonly amplified chromosomes were chromosome 8 and 17. We applied a cut-off for higher gene expression as relative FPKM >1.5. ERBB2 amplifications and high expression were most common (21.5%) of all genes and it was in agreement withHER-2 IHC and FISH result. Among previously reported amplified genes, FGFR1 (5.4%) and PVT1 (8.6%) in chromosome 8, CCND1 , PAK1 (3.2%) and EMSY (4.3%) in chromosome 11, CCNE1 (4.3%) in chromosome 19 were also identified in this study. IGF1R high amplification and expression was found in two samples, and ESR1 , MDM2 , KIT was found in only one sample each. We found uncommon but novel and recurrent highly amplified and expressed genes: CLK4 in 5q (3.2%), AHI / MYB in 6q (3.2%), MMP7 (2.2%) and MALAT1 in 11q (1.1%), and NEK8 in 17q (4.3%) We designed FISH probe for this 5 new genes and confirmed the high amplifications in each sample with FISH. Functional study of these genes will be followed for the driver role of these genes in carcinogenesis and progression of breast cancer cells. Citation Format: Eunshin Lee, Woosung Lim, Kyung-Min Lee, Tae-kyung Yoo, Jongjin Kim, Han-Byoel Lee, Yun-Gyoung Kim, YoungJoon Kang, Min Kyoon Kim, Hyeong-Gon Moon, Dong-Young Noh, Wonshik Han Han. Discovery of novel amplified genes in primary breast cancer with copy number and gene expression analysis of whole exome and transcriptome sequencing data [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 P2-03-18.

Abstract P5-10-10: Effect of prolonged cold ischemic time on immunohistochemical testing of estrogen receptor, progesterone receptor and HER2 expression in breast cancer

Background: Estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2) are the most important predictive and prognostic biomarkers in breast cancer. The American Society of Clinical Oncology/College of American Pathologists recommends that the time from tumor removal to fixation (cold ischemic time) should be kept within 1 hour. Through this study we mean to review the actual cold ischemic time in real practice and analyze whether delayed formalin fixation effects immunohistochemical (IHC) testing results. Methods: Patients, who received surgery for invasive or in situ breast cancer in Seoul National University Hospital, Seoul, Korea between February and December 2013, were retrospectively reviewed. Cold ischemic time was calculated by extracting the time of formalin fixation from the time when surgery ended. All patients were equally divided into two groups (short ischemic group, long ischemic group) according to median cold ischemic time. Chi-square test was done for ER and PR positive/negative (0% negative, ≥1% positive) and student t-test was done for ER and PR percentage. Also χ 2 test was done for HER2 positive/negative and scoring system ranging from 0 to 3+. Results: A total of 615 patients were included in this study. The median cold ischemic time was 2h 43min 4sec (range 6min 36sec – 84h 26min 20sec). Only 48 patients had a cold ischemic time shorter than 1 hour. No association between ER, PR expression and cold ischemic time was found in the χ 2 test (p=0.581, p=0.954) and student t-test (p=0.648, p=0.978). As for HER2 expression, in the long ischemic group, there were significantly more patients with positive immunohistochemical testing results (χ 2 test, p=0.016), and significantly higher grades in HER2 scoring system (χ 2 test, p=0.022). Compared to IHC results, FISH testing for HER2 amplification showed no significant difference (χ 2 test, n=145, p=0.500). This tendency was persisted when patients were divided into four groups by 25 quartile of cold ischemic time. Conclusions: Our findings show that the actual cold ischemic time in practice is longer than recommended guidelines. Despite that, ER, PR expression was not associated with cold ischemic time. As for HER2 expression, longer cold ischemic time was associated with more HER2 positive and higher HER2 score. But this tendency was not showed in FISH testing for HER2 amplification. Citation Format: Tae-Kyung Yoo, Hyeong-Gon Moon, Jisun Kim, Jun Woo Lee, Min Kyoon Kim, Eunshin Lee, Jongjin Kim, Wonshik Han, In-Ae Park, Dong-Young Noh. Effect of prolonged cold ischemic time on immunohistochemical testing of estrogen receptor, progesterone receptor and HER2 expression in breast cancer [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 P5-10-10.

Abstract P2-05-07: Feasibility and sensitivity of fine-needle aspiration biopsies for the detection of somatic mutations using next-generation sequencing in breast cancer

Background/Purpose: Next-generation sequencing (NGS) is being incorporated rapidly into clinical practice. Fine-needle aspiration biopsy (FNAB) specimens have been used feasibly in molecular analysis including direct sequencing and microarrays. They are readily available and enriched in malignant cells, thus providing opportunities for genomic analysis for more clinical samples. In this study, we assessed the feasibility and sensitivity of FNAB for the detection of somatic mutations by NGS compared to bulk tissue. Methods: Bulk tissue and FNAB was sampled via skin superficial to the palpable tumor from surgically resected breast cancer specimen. DNA was extracted from the bulk tissues and FNAB samples obtained from twelve patients. Somatic mutations detected from whole exome sequencing (WES) by next-generation sequencing (NGS) (HiSeq 2500, Illumina) were analyzed for corresponding pairs of bulk tissue and FNAB. Verification of somatic mutations detected exclusively from FNAB and known to be clinically relevant to breast cancer was carried out by Sanger sequencing. Invasive tumor percentages of bulk tissues were evaluated using hematoxylin and eosin (HE San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P2-05-07.

Abstract P1-12-13: Factors associated with adherence to adjuvant endocrine therapy in patients with hormone receptor positive breast cancer

Background/Purpose Adjuvant endocrine therapy in patients with hormone receptor positive breast cancer reduces recurrence and mortality, but many patients are non-adherent to anti-hormonal medication. In order to increase the adherence, it is important to know about factors associated with adherence. So we investigated factors associated with adherence to anti-hormonal medication using variable questionnaires. Methods We carried out a cross-sectional survey of a sample of women who underwent surgery due to breast cancer in the Seoul National University Hospital Breast Care Center from 2007 to 2011 and treated with anti-hormonal medication. Questionnaires were sent to 1,000 patients. The questionnaire booklet included the Medication Adherence Report Scale-5(MARS-5), Women’s Health Questionnaire(WHQ), Beliefs about Medicine Questionnaire(BMQ), Satisfaction with Information about Medicines Scale(SIMS). And to identify patient’s clinical characteristics, we reviewed electronic medical records, retrospectively. Result The response rate of questionnaire was 40.8%(408/1000). Of the answered patients, 263 patients were treated with tamoxifen and 145 patients were treated with aromatase inhibitors(AIs). 197 of 408 answered patients(48.3%) were classified as non-adherence. The rate of non-adherence was 132/263(50.1%) and 65/145(44.8%) in patients treated with tamoxifen and AIs. Of the all answered patients, non-adherent patients had more depressed mood (p Conclusion This study showed associations between depressive mood of breast cancer patients treated with anti-hormonal therapy and adherence. And beliefs and satisfaction with information about medication also associated with adherence. To improve adherence, we should evaluate and correct patient’s mood. And we should provide proper information about medications. Citation Format: Jongjin Kim, Wonshik Han, Hyeong-Gon Moon, Min Kyoon Kim, Eunshin Lee, Tae-Kyung Yoo, Han-Byoel Lee, Young Joon Kang, Yun-Gyoung Kim, Tae Ryung Kim, Dong Young Noh. Factors associated with adherence to adjuvant endocrine therapy in patients with hormone receptor positive breast cancer [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 P1-12-13.