Taiwanese Cancer Research Articles

Abstract 5103: Characterization of the genomic landscape and actionable mutations in Taiwanese prostate cancer patients

Abstract Background: Prostate Cancer (PCa) is the second most common male cancer worldwide and the most common cancer in males over 65 years old. PCa is the sixth most common cancer in Taiwan, and its incidence rate has increased gradually over the past 30 years. PAPR inhibitors are the only type of targeted drug approved by the FDA to treat advanced castration-resistant prostate cancer (CRPC) that has grown after the hormone therapy drugs. This study aimed to characterize the genomic profile of Taiwanese PCa and explore whether there are potential treatment guidances derived. Methods: Formalin-fixed paraffin-embedded (FFPE) tumor tissues from 99 patients diagnosed with PCa and collected at Chi-Mei Medical Center (CMMC) were subject to next-generation sequencing using ACTOnco® CGP panel that cover coding regions of 440 cancer-related genes. Genomic alterations (GAs) and signatures, including single nucleotide variations (SNVs), short insertions and deletions (InDels), copy-number variations (CNVs), fusion genes, tumor mutations burden (TMB), and microsatellite instability (MSI) status were retrieved. The gene mutation prevalence was compared with the Western population using the TCGA prostate cancer dataset. Results: A total of 1558 variants was identified, including 215 SNVs, 22 InDels, 1178 CN gain and 135 homozygous deletions. Similar to the Western population, the most frequently mutated genes were SPOP (21.2%), KMT2C (10.1%), MUC16 (8.1%), TP53 (6.1%), ARID2 (5.1%), IDH1 (5.1%), PTGS2 (5.1%). Further analysis showed that 12 patients harbored deleterious/likely deleterious mutations in genes implicated in homologous recombination (HRR) DNA repair pathway, including three BRCA2, two CDK12, BRAD1 and ARID1A, respectively, and one ATR, ATM and PALB2, respectively. All BRCA2 mutations were predicted to be germline origin and only one-third of 12 HRR mutants had a biallelic gene loss. Median TMB calculated from NGS data is 0.6 muts/Mb (range 0 to 4.5) and all samples are microsatellite-stable (MSS). Therapeutically actionable gene and pathway analysis revealed that up to 24.2% of patients harbored at least one aberration against FDA-approved targeted drugs, such as MET copy number (CN) gain (11.1%), mTOR pathway aberration (6.1%), HRR gene biallelic loss (4%), BRAF activating mutation (4%), IDH1 activating mutations (4%), EGFR CN gain (3.0%), FGFR1 CN gain (2.0%), MET fusion (1.0%) and BRAF fusion (1%). Conclusion: This CGP study unraveled the genomic profile of Taiwanese prostate cancers, and the results showed that GAs identified could serve as a potential biomarker to guide personalized, targeted therapies in one-fourth of patients. Citation Format: Kien Thiam Tan, Chia-Ling Wu, Yen-Jung Lu, Ka-Po Tse, Chien-Feng L Li. Characterization of the genomic landscape and actionable mutations in Taiwanese prostate cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5103.

Differential whole-genome doubling and homologous recombination deficiencies across breast cancer subtypes from the Taiwanese population

Whole-genome doubling (WGD) is an early macro-evolutionary event in tumorigenesis, involving the doubling of an entire chromosome complement. However, its impact on breast cancer subtypes remains unclear. Here, we performed a comprehensive and quantitative analysis of WGD and its influence on breast cancer subtypes in patients from Taiwan and consequently highlight the genomic association between WGD and homologous recombination deficiency (HRD). A higher manifestation of WGD was reported in triple-negative breast cancer, conferring high chromosomal instability (CIN), while HER2 + tumors exhibited early WGD events, with widely varied CIN levels, compared to luminal-type tumors. An association of higher activity of de novo indel signature 2 with WGD and HRD in Taiwanese breast cancer patients was reported. A control test between WGD and pseudo non-WGD samples was further employed to support this finding. The study provides a better comprehension of tumorigenesis in breast cancer subtypes, thus assisting in personalized treatment.

Abstract 2481: Concordance of PAM50 molecular subtyping between oligonucleotide microarray and nanoString nCounter assay for Taiwanese breast cancer

Abstract Concordance of PAM50 molecular subtyping between oligonucleotide microarray and nanoString nCounter assay for Taiwanese breast cancer Introduction Breast cancer is molecularly heterogeneous and at least five molecular subtypes have been defined according to intrinsic genes transcription. Currently microarrays, RT-PCR, and next generation RNA sequencing have been adopted for subtyping in addition to the original nanoString nCounter assay. This study aimed to evaluate the consistency in subtyping Taiwanese breast cancers. Methods A total of 109 Taiwanese breast cancers (24 with adjacent normal breast) were assayed with Affymetrix Human Genome U133 plus 2.0 arrays and 96 were assayed with the customized 145-gene nanoString Codeset BCeCSig, of which the academic PAM50 was investigated. Of note, 64 patients were assayed for both platforms. Subtyping with the nearest centroid (single sample prediction) was called for each patient; Spearman's rank correlation coefficients were used, and samples were unclassified if correlation coefficients to all 5 centroids were less than 0.1. Results Table 1 showed the distributions of molecular subtypes with clinical IHC results. For 24 matched normal breast tissues, 21 were classified as normal breast-like. For 64 breast cancers (one unclassified) tested for both platforms, 41 (65%) were classified with identical subtype, resulting in a fair Kappa statistic of 0.55. Taking nCounter as gold standard, classification accuracy was 50% (3/6), 72% (13/18), 53% (10/19), 87% (13/15), and 40% (2/5) for basal-like, HER2-enriched, luminal-A, luminal-B, and normal breast-like subtype using Affymetrix microarray. ER-/HER2- phenotype and basal-like subtype experienced worse relapse free survival at 8-year follow-up (55.6% and 63.6% censored). Table 1.Molecular subtypes and IHC phenotypes (two unclassified and one missing IHC result).Subtype IHCBasal-likeHER2-enrichedLuminal-ALuminal-BNormal breast-likeHR+/HER2+276174HR+/HER2-6442429HR-/HER2+625209HR-/HER2-144003 Conclusions: Using public domain microarrays as a handy shortcut for breast cancer molecular subtyping is luring in scientific communities. Our experiments showed that fundamental discrepancy existed between distinct approaches, and more cation should be taken without too much extrapolation. On the other hand, adjacent normal breast tissue did display normal breast-like character, and this subtype should be viewed as a continuum of surrounding stroma. Citation Format: Chi-Cheng Huang, Chao-Chiang Tu, Mong-Hsun Tsai, Liang-chuan Lai, Ching-Shui Huang. Concordance of PAM50 molecular subtyping between oligonucleotide microarray and nanoString nCounter assay for Taiwanese breast cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2481.

Abstract 2503: Molecular profiling of Taiwanese breast cancers reveals potential therapeutic targets: Preliminary results of 160 subjects

Abstract Introduction: Breast cancer is one of the leading causes of cancer-related death in women, and the annual incidence of breast cancer has continuously increased throughout the past two decades in Taiwan. There is a demand in developing an Asian-based genetic profiling database for breast cancer in improving the treatment response and recurrence rate. This study aimed to determine the difference of molecular alternations and identify the potential biomarkers regarding therapeutic target by analyzing the genetic profiling from a large cohort of breast cancer subjects using the method of next-generation sequencing (NGS). Materials and methods: The protocol consisted of three years enrollment and approximately four years follow-up after enrollment. Individual subject was assigned into Group 1 [planned to received surgery as first-line treatment and followed by adjuvant therapy, Group 2 [planned to receive neoadjuvant therapy as the first-line treatment and followed by surgery], and Group 3 [diagnosed with de novo and treatment naïve stage IV breast cancer, or stage IV breast cancer with recurrence beyond three years after surgery]. Molecular profiling and potential biomarkers were determined using Thermo FisherTM OncomineTM Comprehensive Assay version 3 (TMO comprehensive assay) from Formalin-Fixed Paraffin-Embedded (FFPE) tissues. Results: We presented the initial 160 subjects as preliminary results. The TMO comprehensive assay is a targeted, NGS assay that enables the detection of relevant SNVs, CNVs, gene fusions, and indels from 161 unique genes to help inform drug discovery research and clinical trial research programs such as the NCI-MATCH trial. Level of actionability of sequenced targets were evaluated with ESMO Scale of clinical actionability of molecular targets (ESCAT). Of note, Tier I [alteration-drug match associated with improved outcome in clinical trials], Tier II [antitumor activity associated with the matched alteration-drug but lack of prospective outcome data], and Tier III [matched drug-alteration that led to clinical benefit in another tumor type] matched targets including ERBB2 amplification, BRCA1/2 germline mutation, PIK3CA mutation, NTRK translocation (Tier I), PTEN loss, ESR1 mutation, AKT1 mutation, ERBB2 mutation (Tier II), and BRCA1/2 somatic mutation, MDM2 amplification, and ERBB3 mutation (Tier III) were retrieved and analyzed. 108 out of 160 sequenced subjects (67.5%) reported as least one variant (average 1.7 per case, ranged from 1 to 10). Two group 3 heavily pre-treated patients were identified to have all actionable genes varied. Ipatasertib, alpelisib, and talazoparib, for instance, had been advocated for the corresponding PTEN p.(Q219*), PIK3CA p.(G118S), and BRCA2 p.(Q1107*) mutations. Conclusion: Targeted sequencing of actionable genes is believed to provide clinical applicability and substantial benefits for Taiwanese breast cancer patients. A valid scale of clinical actionability should be adopted for precision medicine practice under multidisciplinary molecular tumor board. Further details will be updated when more samples are ready for sequencing and actionability analyses in coming years. Citation Format: Chi-Cheng Huang, Chun-Yu Liu, Ling-Ming Tseng. Molecular profiling of Taiwanese breast cancers reveals potential therapeutic targets: Preliminary results of 160 subjects [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2503.

Abstract P4-07-12: Residual risk stratification of Taiwanese breast cancer following curative therapies with digital RNA counting and actionable mutations sequencing

Abstract Introduction: Breast cancer is the most common female malignancy in Taiwan and treatment outcomes have improved enormously with early detection and advancements in adjuvant therapy. Unfortunately, a subset of early stage breast cancer still suffers from recurrence, metastasis, or even cancer-specific death despite treatment improvement, and conventional pathological factors fail to provide sufficient explanatory power accounting for the observed prognostic discrepancy. The aim of the project is to take advantage of digital RNA counting for gene expression and targeted sequencing of actionable mutations to develop a predictive model for operable breast cancers initially treated with curative intention, and the risk of recurrence, distant metastasis, as well as breast cancer-specific mortality will be speculated. Methods: Two approaches are applied for candidate genes yield; we have identified concurrent genes from the coherent patterns between genomic and transcriptional profiles, and the synthesized signatures were prognostic. In addition, concordant genes from leading edge analysis of published gene expression signatures were retrieved, and the consensus of leading edge subsets could stratify breast cancer patients into groups with distinct survival patterns. With bioinformatics tools, the harvested genes will be used to build a risk stratification model, and the residual risk after surgery and adjuvant therapy will be evaluated from archived formalin-fixed paraffin embedded (FFPE) cancer tissues, augmenting the clinical significance of the proposed signature. And finally, actionable mutations will be identified for relapsing patients through targeted sequencing. Results: There were 1584 extended concurrent genes with the threshold MAD (medium absolute deviation) of 0.9, constituting 15.89% of common genes across copy number variation and gene expression arrays. We also constructed breast cancer risk prediction using concordance of genes from leading edge analysis of various prognostic signatures; enrolled signatures were split into either the up-regulated or down-regulated gene sets within each study, and leading edge analysis of filtered up-/down-regulated gene sets from all training data sets were performed, followed by the PLS (partial least square) risk predictive model composed of consensus of leading edge subsets among all assayed subjects. The 145-gene BCeC Sig platform designated for the current study has been established, and was prognostic in 96 Taiwanese breast cancers (log-rank test: P<0.01). Among 37 breast cancers categorized into the high-risk group, there were 383 SNPs, 23 deletes, and 11 insertions. Variations per breast cancer were between 91 and 162 (average: 120). When collapsed into genes, the most altered genes were: BRCA1/2 (30 variants), FANCA(27), NOTCH(21), ALK, AR (20), ATM, BSG(14), FGFR, MET(12), JAK2, PDFGR, RET(11), CSF1R, DAB2I, and TP53(10). Discussion and conclusions: Archived pathological specimens provide an invaluable source to validate the residual risk prediction model for breast cancer patients managed with contemporary multi-modality treatment including surgery, chemo-, endocrine, and targeted therapy. Degraded nucleic acid from FFPE samples could be tackled by the latest NanoString digital RNA counting. This breast cancer residual risk model, composed of concurrent genes and consensual leading edge subset of published gene expression signatures, is believed to provide clinical applicability and substantial benefits for Taiwanese breast cancer patients in terms of personalized medicine Citation Format: Chi-Cheng Huang, Ching-Shui Huang. Residual risk stratification of Taiwanese breast cancer following curative therapies with digital RNA counting and actionable mutations sequencing [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P4-07-12.

Interaction of Interleukin-16 Genotypes With Betel Quid Chewing Behavior on Oral Cancer in Taiwan.

Interleukin-16 (IL-16) is reported to play an important role in inflammation, carcinogenesis and tumoricidal processes, however, the contribution of IL-16 genotype to oral carcinogenesis is still largely unrevealed. Thus, the study aimed to investigate the contribution of IL-16 genotypes to Taiwan oral cancer risk. The genotypes of IL-16 rs4778889, rs11556218, and rs4072111 were revealed among 958 oral cancer cases and 958 control subjects by polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP). First, the distributions of genotypic (p=0.0004) and allelic (p=0.0001) frequencies of IL-16 rs11556218 were significantly different between the case and control groups. In detail, the frequencies of IL-16 rs11556218 TG and GG were 28.1 and 5.8%, respectively, among oral cancer patients, significantly higher compared to those among controls (25.0% and 2.7%, respectively). Second, no difference was observed regarding IL-16 rs4778889 or IL-16 rs4072111. Last, there was a synergistic effect of betel quid chewing behavior and risky IL-16 rs11556218 genotype on oral cancer risk. The study indicates that the IL-16 rs11556218 G allele synergistically interacts with betel quid chewing behavior, contributing to increased risk of oral cancer in Taiwanese.

Association of Interleukin-4 Polymorphisms With Breast Cancer in Taiwan.

The present study aimed at evaluating the contribution of IL-4 promoter T-1099G (rs2243248), C-589T (rs2243250), C-33T (rs2070874) genotypes to the risk of breast cancer in Taiwanese. A total of 1232 breast cancer patients and 1232 age-matched controls were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methodology. Genotypic frequencies of IL-4 rs2243248, rs2243250 and rs2070874 were not differentially distributed between case and control groups. Consistently, there was no difference in the distribution of allelic frequencies among patients and controls. IL-4 rs2243248, rs2243250 and rs2070874 do not confer breast cancer susceptibility in Taiwanese.

The Performance of Different Artificial Intelligence Models in Predicting Breast Cancer among Individuals Having Type 2 Diabetes Mellitus.

Objective: Early reports indicate that individuals with type 2 diabetes mellitus (T2DM) may have a greater incidence of breast malignancy than patients without T2DM. The aim of this study was to investigate the effectiveness of three different models for predicting risk of breast cancer in patients with T2DM of different characteristics. Study design and methodology: From 2000 to 2012, data on 636,111 newly diagnosed female T2DM patients were available in the Taiwan’s National Health Insurance Research Database. By applying their data, a risk prediction model of breast cancer in patients with T2DM was created. We also collected data on potential predictors of breast cancer so that adjustments for their effect could be made in the analysis. Synthetic Minority Oversampling Technology (SMOTE) was utilized to increase data for small population samples. Each datum was randomly assigned based on a ratio of about 39:1 into the training and test sets. Logistic Regression (LR), Artificial Neural Network (ANN) and Random Forest (RF) models were determined using recall, accuracy, F1 score and area under the receiver operating characteristic curve (AUC). Results: The AUC of the LR (0.834), ANN (0.865), and RF (0.959) models were found. The largest AUC among the three models was seen in the RF model. Conclusions: Although the LR, ANN, and RF models all showed high accuracy predicting the risk of breast cancer in Taiwanese with T2DM, the RF model performed best.

Recapitulation of inflammatory and immune-evasive subtypes of oral cancer cells in immunodeficient mice.

e14199 Background: Prior study stratified Taiwanese oral cancer specimens (n = 40) and cell lines (n = 7) into three distinct molecular subtypes, namely classical (CL), mesenchymal (MS), and basal (BA). In a cell line derived xenograft (CDX) model, we observed MS grew at least 10-fold slower than CL did in immunodeficient mice. By using RNA-seq to portrait the transcriptomes of human tumor and mouse stroma simultaneously, contribution of mouse innate immunity in restricting the growth of human oral cancer cells was assessed. Methods: A half millions of mycoplasma-free OC3 (MS) or TW2.6 (CL) cells with matrigel were subcutaneously implanted into the flank of 10 to 16 weeks old NOG (NOD/SCID/Il2rgtm). RNAs of CDX tissues from OC3-NOG (n = 7) and TW2.6-NOG (n = 5) were extracted and subjected to stranded mRNA sequencing. Clean reads were aligned to GRCh38 (human) and GRCm38 (mouse), respectively, followed by identifying differentially expressed genes and gene set enrichment analysis. Results: Up-regulation of signature genes for dendritic cells and macrophages and enrichment of innate immunity, including Tnfa-Nfkb signaling, interferon alpha response, interferon gamma response and inflammation were detected in OC3- but not in TW2.6-CDXs. These results suggested that OC3 (MS) was more immunogenic than TW2.6 (CL), which is reminiscent of the inflammatory MS (IMS) subtype of head and neck cancer recently described by Keck et al (n = 938, Clin Cancer Res 2015, 21: p870. PMID: 25492084). Conclusions: By RNA-seq/xenome analysis of CDXs, we provide evidence that compared to CL, MS subtype of oral cancer cells elicited a stronger innate immunity in NOG mouse. Alternatively, CL subtype might have evolved as a prototype with superiority in immune escape.

The Contribution of Interleukin-12 Genetic Variations to Taiwanese Lung Cancer.

Lung cancer is the leading cause of cancer-related death and a better marker for advanced personalized therapeutic approaches, such as immunotherapies, is in urgent need. Interleukin-12 (IL-12) is a cytokine that has been reported to exhibit potent tumoricidal effects, however, the contribution of IL-12 genotypes to lung cancer is still largely unrevealed. The aim of this study was to investigate whether single nucleotide polymorphisms (SNPs) in IL-12A and IL-12B are associated with lung cancer in a Taiwanese population. Genotypes of 358 lung cancer patients and 716 controls were determined by the polymerase chain reaction-restriction fragment length polymorphism method. The distributions of genotypic (p=0.0036) and allelic (p=0.0005) frequencies of IL-12A rs568408 demonstrated significant differences between cases and controls. In detail, the AA genotype of IL-12A rs568408 was associated with a significantly elevated risk of lung cancer compared with the GG genotype (odds ratio(OR)=2.41, 95% confidence interval(CI)=1.36-4.29, p=0.0021). No difference was observed regarding IL-12A rs2243115 and IL-12B rs3212227 genotypes between the case and control groups. In addition, the results of interaction analysis showed that the AA genotype of IL-12A rs568408 was associated with elevated lung cancer risk, especially among those with smoking habits (p=0.0043). IL-12A rs568404 AA genotype may contribute to the etiology and serve as a genomic determinant of lung cancer in Taiwanese, especially smokers.

Abstract 2587: Imbalances in the disposition of estrogen and naphthalene in Taiwanese breast cancer patients

Abstract The aims of this study were to measure the background level of quinone metabolites of 17β-estradiol (E2), including E2-2,3-Q and E2-3,4-Q, and naphthalene quinone (1,2-NPQ and 1,4-NPQ)-derived protein adducts in human albumin (Alb) derived from breast cancer patients (n=143) and healthy controls (n=122) in Taiwanese women and to explore the relationships between these adducts with risk factors of breast cancer. Results from the analysis indicated that Alb adducts of estrogen quinones, including E2-2,3-Q-4-S-Alb and E2-3,4-Q-2-S-Alb were detected in all subjects with mean levels at 400 and 676 pmole/g in cancer patients and 204 and 67 pmole/g in controls, respectively. In addition, the mean levels of Alb adducts of 1,2-NPQ and 1,4-NPQ on human serum albumin were estimated to be 209 and 73.6 pmole/g in cancer patients and 113 and 174 pmole/g in controls, respectively. Levels of E2-2,3-Q-4-S-Alb correlated significantly with those of E2-3,4-Q-2-S-Alb. Similarly, levels of 1,4-NPQ-Alb correlated significantly with those of 1,2-NPQ-Alb. Additionally, we noticed that E2-2,3-Q-4-S-Alb and E2-3,4-Q-2-S-Alb positively correlated with 1,4-NPQ-Alb (p<0.001) whereas levels of E2-2,3-Q-4-S-Alb associated with 1,2-NPQ-Alb (p<0.01). Furthermore, we observed that ratio of E2-3,4-Q-2-S-Alb to E2-2,3-Q-4-S-Alb in breast cancer patients were ~5 fold greater than those of healthy controls whereas ratio of 1,2-NPQ-Alb to 1,4-NPQ-Alb in cancer patients were ~4 fold greater than those of healthy controls. This evidence suggests that the cumulative body burden of estrogen quinones is a significant indicator of the risk of developing breast cancer and that breast cancer risk is associated with the imbalances in the disposition of estrogen and naphthalene in Taiwanese women. Citation Format: Po-Hsiung Lin, Dar-Ren Chen, Wei-Chung Hsieh, Mao-Huei Ko, Yi-Lun Liao. Imbalances in the disposition of estrogen and naphthalene in Taiwanese breast cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2587.

Abstract 5370: Targeted sequencing of actionable mutations to enhance the prognostication of concurrent genes-defined high-risk breast cancers

Abstract Purpose: The aim of the study is to perform targeted sequencing to enhance the prognostic power of breast cancer concurrent genes. Somatic mutations with clinical significance will be identified. Materials and Methods: Our previous study had identified the concurrent genes signature from the coherent patterns between genomic and transcriptional profiles among Taiwanese breast cancers, and those predicted into the high-risk group were furthered assayed by targeted sequencing. Target-enrichment sequencing was performed with Illumina SolexaTM technology with read length of 150 bp and was analyzed with Agilent SureCallTM software. Results: In total, 109 breast cancer samples were collected during surgery and analyzed with AffymetrixTM U133 plus 2.0 microarrays. Breast cancer risk prediction was performed with the published concurrent genes signature. Targeted sequencing of 61 high-risk breast cancers revealed 1061 variants, including 76 pathogenic and 545 likely pathogenic variants based on ACMG classification. The most impacted genes were NOTCH, BRCA1, AR, ERBB2, FANCA, ATM, and BRCA2, harboring 57, 36, 30, 27, 27, 26, and 26 variants. Table 1 showed that the most frequent pathogenic deletions were FGFR1 , ATM, and WT1 (47, 47, and 37 patients) while BRCA1 (rs1799965, non-sense mutation), FGFR2 (missense mutation), and BRCA1 (rs1799949, non-sense mutation) were the most frequent pathogenic SNPs (44, 35, and 11 breast cancers, respectively). The distributions of deletions and pathogenic SNPs were similar across ER+, HER2+, and triple-negative breast cancers. Conclusion: This two-stage predictive model, concurrent genes signature risk stratification followed by targeted sequencing of actionable genes, is believed to provide clinical applicability and substantial benefits for Taiwanese breast cancer patients. Table 1. Pathogenic mutations affecting more than 10 high-risk breast cancers.GeneMutation typeEffectAffected subjectsPathwayPotential therapyFGFR1DeletionFRAME_SHIFT|NM_00117406347PI3K, FGF signalingDovitinibATMDeletionFRAME_SHIFT|NM_00005147DNA damage responseATR inhibitor, PARP inhibitor, Chk 1/2 inhibitorWT1DeletionFRAME_SHIFT|NM_00037837Mutated in Wilm's tumorPDGFRADeletionFRAME_SHIFT|NM_00620618PDGF signalingSorafenib, imatinib, sunitinibPIK3RDeletionFRAME_SHIFT|NM_18152316PI3K signalingBuparlisibRUNX1DeletionFRAME_SHIFT|NM_00100189015Transcription factor, mutated in AMLBRCA1 (rs1799965)SNPnonsense44DNA damage responsePARP inhibitor, cisplatinFGFR2 (chr10:123298158:T:C )SNPmissense35PI3K, FGF signalingBrivanib, nintedanib, dovitinibBRCA1 (rs1799949)SNPnonsense11DNA damage responsePARP inhibitor, cisplatin Citation Format: Chi-Cheng Huang, Chao-Chiang Tu, Ching-Shui Huang. Targeted sequencing of actionable mutations to enhance the prognostication of concurrent genes-defined high-risk breast cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5370.

Abstract P5-07-04: Nuclear miR-133 expression is associated with breast cancer progression and survival

Abstract Background:MiRNAs (microRNAs) can function as the tumor suppressor gene or oncogene to affect carcinogenesis, therapy response and progression in human cancer. The prevailing view is that miRNAs function to regulate mRNA stability and translation in the cytoplasm. However, multiple studies have detected miRNAs in the nuclear compartment. Nuclear miRNAs have been proven to regulate the functions of nuclear protein and synthesis of nuclear miRNA. We have found circulating muscle-enriched miR-133a can serve as prognostic biomarkers for breast cancer. Herein, we aim to explore the subcellular distribution, prognostic significance and treatment response of miR-133a in Taiwanese breast cancer. Methods: Circulating miR-133a levels were conducted with the serum from 188 patients prior to surgery and from 82 healthy controls. The subcellular distribution patterns of miR-133a were analyzed by Chromogenic In Situ Hybridization (CISH) in paraffin embedded tissue sections from 224 cases. Cell viability and cell cycle were determined by XTT assay and flowcytometry. Protein and mRNA levels were examined by western blotting and RT-qPCR (TaqMan) assay. Results: Our results showed that circulating miR-133a levels of breast cancer patient are significantly higher than those of health subjects, suggesting that increased miR-133a secretion from unknown tissues or cells may promote carcinogens in Taiwanese breast cancer. Using the receiver operating characteristic (ROC) curve to examine the diagnostic value of circulating miR-133a in breast cancer (AUC=0.834), we found that increased miR-133a levels could serve as a diagnostic biomarker in accessing Taiwanese breast cancer risk. Nevertheless, CISH analysis revealed that nuclear miR-133a frequency was inversely correlated with tumor grade, stage and lymph node metastasis (p=0.029, 0.012, and 0.0008), and cytoplasmic miR-133a intensity was positively correlated with recurrence (p=0.010). Furthermore, high frequency of nuclear miR-133a was correlated with a better overall survival rate (p=0.039) and improved the survival rate of patients who received hormone therapy (p=0.009). To further investigate the role of nuclear miR-133a, an additional miR-133a, denoted as Nu-miR-133a, ending in nuclear localization element was constructed and found to be highly enriched in the nucleus. Ectopic expression of wild-type and Nu-miR-133 inhibited proliferation of breast cancer cells. Both wild type miR-133a and Nu-miR-133a increased S phase arrest as well as the levels and foci of γ-H2AX. Unexpectedly, we found that wild type miR-133a decreased ATM expression but activated p38 MAPK. Notably, H2AX can be also phosphorylated by p38 MAPK kinase in addition to ATM and ATR. The underlying mechanism between nuclear miR-133a and potentially p38-mediated DNA damage response (DDR, γ-H2AX and S phase arrest) will be needed further elucidation. Conclusions: In conclusion, we provide the first evidence that nuclear miR-133a may play a crucial role in the development and treatment response of breast cancer potentially through its mediated DDR. The interplay among nuclear miR-133a, p38 and DDR require further investigations. Citation Format: Hou M-F, Chen F-M, Yang S-F, Dai H-Y, Yeh Y-T. Nuclear miR-133 expression is associated with breast cancer progression and survival [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P5-07-04.

Is vegetarian diet associated with a lower risk of breast cancer in Taiwanese women?

BackgroundStudies on the relationship between vegetarian diet and breast cancer in Asian populations are limited. This study aimed to investigate the relationship between vegetarian diet, dietary patterns, and breast cancer in Taiwanese women.MethodsThis case-control study compared the dietary patterns of 233 breast cancer patients and 236 age-matched controls. A questionnaire about vegetarian diets and 28 frequently-consumed food items was administered to these 469 patients in the surgical department of Taipei Tzu Chi Hospital. Serum biochemical status was also examined.ResultsThere were no significant differences between the two groups for age, education, family history, oral contraceptive usage, or regular exercise. However, the cancer group presented with both a higher body mass index and an older age of primiparity (P < 0.05). Two food items (shellfish and seafood) were highly correlated (correlation coefficient = 0.77), so shellfish was excluded to avoid multicollinearity. A factor analysis of 27 food items produced five dietary patterns: meat, processed meat, fruit/vegetable/soybean, dessert/sugar, and fermented food. Multivariate logistic regression showed that meat/fat and processed meat dietary patterns were associated with breast cancer risk (odds ratio (OR): 2.22, 95% CI 1.67–2.94, P < 0.001; OR: 1.49, 95% CI 1.09–2.04, P = 0.013, respectively). Vegetarian diet, high isoflavone intake, and high albumin levels were inversely associated with breast cancer risk (P < 0.05). Vegetarians had a higher daily soy isoflavone intake than non-vegetarians (25.9 ± 25.6 mg vs. 18.1 ± 15.6 mg, P < 0.001).ConclusionsVegetarian diets show as protective role against breast cancer risk, while meat and processed meat dietary patterns are associated with a higher breast cancer risk.

The Contribution of Matrix Metalloproteinase-7 Promoter Genotypes in Breast Cancer in Taiwan.

The matrix metalloproteinase (MMP) family of enzymes are in charge of degradation of various components of the extracellular matrix and their functional genetic polymorphisms may be associated with cancer susceptibility. The functional polymorphisms in the promoter region of MMP7 (A-181G and C-153T) have been reported to influence the binding capacity of nuclear proteins and may contribute to genetic susceptibility to cancer. In this study, we focused on investigating the contribution of the genotypes of MMP7 (A-181G and C-153T) to breast cancer in Taiwan. These two polymorphisms were genotyped in 1,232 patients with breast cancer and 1,232 controls by polymerase chain reaction-restriction fragment length polymorphism methodology. The odds ratios (ORs) after adjusting for age, family history of cancer, smoking and alcohol drinking status for those carrying AG and GG genotypes at MMP7 promoter A-181G were 1.22 (95%CI=0.91-1.63, p=0.2235) and 2.84 (95%CI=1.64-7.48, p=0.0007) respectively, compared to those carrying the wild-type AA genotype. Supporting this finding, the adjusted OR for those carrying the G allele at MMP7 promoter A-181G was 1.57 (95%CI=1.29-1.93, p=0.0008), compared to those carrying the wild-type A allele. There was no polymorphic genotype at MMP7 C-153T found among any of the investigated individuals. Our findings suggest that the MMP7 A-181G polymorphisms may play a role in determining personal cancer susceptibility and GG genotype at MMP7 A-181G may serve as a biomarker for early detection and prediction of breast cancer in Taiwanese.

Targeted gene sequencing panel for identifying actionable genomic alterations in Taiwanese lung cancer patients.

e20522 Background: Lung cancer boasts an arsenal of targeted therapies directed at various molecular aberrations such as EGFR mutations and fusion genes. Simultaneous assessment for genetic alterations provides biomarkers to assist clinicians in their treatment selections. Methods: A total of 43 FFPE (formalin-fixed, paraffin-embedded) samples obtained from lung cancer patients in Taiwan were subjected to next-generation sequencing (NGS), using a compact panel encompassing 17 potentially actionable, lung cancer-associated genes. NGS was performed on the Ion Torrent PGM or Proton System with a targeted average depth of &gt; 800x. Genomic alterations detected were categorized as single nucleotide variants (SNV) or chromosomal copy number alterations (CNA). Results: A total of 74 putatively actionable genomic alterations were detected across 38 patients (88.4%), which may predict sensitivity or resistance to established and/or therapies that are still in clinical development. 19 patients (44.2%) had tumors harboring activating EGFR mutations in the tyrosine kinase (TK) domain – L858R substitution or exon 19 deletion, which may suggest clinical benefit to first- and second-generation EGFR TK inhibitors (TKIs). Three patients (7.0%) whose tumor had acquired the EGFR T790M mutation were advised to switch to third-generation EGFR TKI. Possible resistance mechanisms to EGFR TKIs via increased copy number of EGFR (14.0%, n = 6) and MET (11.6%, n = 5) were observed. Deletion or loss of CDKN2A gene were detected in 14 patients (32.6%), which may confer sensitivity to CDK4/6 inhibitors. Inhibition of downstream effectors in the RAS/RAF/MEK/ERK and PI3K/AKT/mTOR pathways were suggested for patients whose tumor harbored PIK3CA and/or KRAS oncogenic mutations. Conclusions: Next-generation sequencing using a compact panel of genes may be sufficient to identify biomarkers for targeted therapies selection for most lung cancer patients in Taiwan.

Risk of Second Primary Cancer in People with Non-melanoma Skin Cancer: A Nationwide Cohort Study.

PurposePrevious western studies have found Caucasians with skin cancer, either melanoma or nonmelanoma skin cancer (NMSC), have an elevated risk of second primary cancer. Our objective was to assess the risk of second primary cancer in Taiwanese with NMSC.Materials and MethodsBy using data from Taiwan’s National Health Insurance Research Database, we conducted a population-based cohort study to assess the risk of incident second primary cancer in Taiwanese affected by NMSC.ResultsWe identified 505 subjects with NMSC and 2,020 matched controls. After adjustment for potential confounders including age, sex, urbanization, and Charlson Comorbidity Index, people who had NMSC had a 1.43-fold (95% confidence interval [CI], 1.05 to 1.96) risk for the development of second primary cancer as comparedwith control group. Menwith NMSC had a 2.99-fold (95% CI, 1.00 to 9.10) risk for second primary cancer involving the lip, oral cavity, and pharynx and a 3.51-fold (95% CI, 1.21 to 10.17) risk for second primary cancer involving the genitourinary organs when compared to the control group. By contrast, women with NMSC did not have an increased risk of second primary cancer.ConclusionThis study revealed Asians with NMSC have an increased risk of second primary cancer. Our findings can be a useful reference for health care for people diagnosed with NMSC.

Body mass index and breast cancer: analysis of a nation-wide population-based prospective cohort study on 1 393 985 Taiwanese women.

Background:Asian women have a younger age at onset of breast cancer and a lower body mass index (BMI) than Western women. The link between obesity and risk of breast cancer in Asian women is still elusive. We aimed to investigate the effect of BMI on the risk of incident breast cancer in Taiwanese women.Methods:A total of 1 393 985 women who had been cancer-free before recruitment and attended a nation-wide Taiwanese breast cancer-screening program between 1999 and 2009 were enrolled using a prospective cohort study. Obesity and other relevant variables (such as menopause status and other biochemical markers) were collected through in-person interviews, anthropometric measurements and blood samples at first screen. Incident breast cancers during follow-up were ascertained through the linkage of the cohort with the National Cancer Registry and the National Death Certification System.Results:A total of 6969 and 7039 incident breast cancer cases were identified among women enrolled before and after menopause, respectively. Compared with a BMI range of 18.5–23.9 kg m−2, the incremental level of BMI in the enrolled women before menopause revealed a lack of statistically significant association with the risk of incident breast cancer (adjusted hazard ratio=0.94, 0.98, 1.02, 1.01 and 0.82 for BMI <18.5, 24–26.9, 27–29.9, 30–34.9 and ⩾35, respectively), but the incremental level of BMI in the enrolled women after menopause led to a statistically significant incremental increase in the risk of breast cancer (adjusted hazard ratio=0.78, 1.19, 1.31, 1.53 and 1.65 for BMI <18.5, 24–26.9, 27–29.9, 30–34.9 and ⩾35, respectively) after adjusting for other explanatory risk factors.Conclusion:Obesity acts mainly as an influential promoter of the development of late-onset breast cancer after menopause in Taiwanese women.

Abstract 3071: MicroRNA-486 (miR-486) promotes invasion by targeting RECK in nasopharyngeal carcinoma cells

Abstract Nasopharyngeal carcinoma (NPC) is a common cancer in Taiwanese and people living in the southeastern region of China. In order to investigate NPC pathogenesis, our laboratory has previously established NPC-TW01 cell line, which was derived from a Taiwanese NPC patient. We also established a NPC-TW01N1 cell line, which was derived from the NPC xenograft in SCID mouse. Both NPC-TW01 and NPC-TW01N1 cell lines have an identical DNA profile (STR). NPC-TW01N1 cell line has higher malignant potency than NPC-TW01 line. For example, the proliferation and invasion capabilities of NPC-TW01N1 line were higher than that of NPC-TW01 line. When we used microRNA microarray to compare the endogenous microRNA expression profiles between NPC-TW01 and NPC-TW01N1 lines, the miR-486, miR-200a and other miRNAs showed higher expression in NPC-TW01N1 cell. Transfection of exogenous miR-486 or anti-miR-486 into NPC-TW01 cells or NPC-TW01N1 cells revealed that miR-486 involves in the regulation of NPC cell progression and invasion. To determine whether miR-486 targeted gene involves in progression and invasion, we analyzed microRNA targeted gene database, Ingenuity pathway databases, and cDNA expression microarrays from miR-486 transfected cells. We found that the RECK gene is one of miR-486 candidate targeted genes that involves in NPC cell progression and invasion. We used pmirGLO luciferase reporter vector assays to demonstrate that miR-486 directly targets RECK 3′ UTR. When we transfected miR-486 or RECK siRNA into NPC cells, we found that they can accelerate tumor progression and invasion. It is concluded that miR-486 and RECK genes maybe potential targets for NPC therapy. Citation Format: Chin-Tarng Lin, Cheng-Der Wu. MicroRNA-486 (miR-486) promotes invasion by targeting RECK in nasopharyngeal carcinoma cells. [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 3071. doi:10.1158/1538-7445.AM2015-3071

Weekly-docetaxel chemotherapy for Taiwanese patients withmetastatic castrate-resistant prostate cancer

Weekly-docetaxel chemotherapy for Taiwanese patients withmetastatic castrate-resistant prostate cancer