Positive Subtypes Research Articles

Preliminary study on identification of estrogen receptor-positive breast cancer subtypes based on dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) texture analysis.

Currently, breast cancer is divided into Luminal A, Luminal B, HER-2 overexpression (HER-2) and basal cell at genetic level. However, the differential diagnosis of estrogen receptor (ER)-positive breast cancer subtypes is rare. Therefore, we aimed to investigate the feasibility of identifying the ER-positive breast cancer subtypes based on dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) texture analysis. A retrospective analysis was performed for clinical data of 51 patients with ER-positive breast invasive ductal carcinoma confirmed by surgery and pathology from January 20 to October 2018. FireVoxel texture analysis software was used to delineate the tumor boundary layer by layer. The differences in the above characteristics between Luminal A and Luminal B breast cancer were compared, and the diagnostic efficacy of statistically significant texture parameters for ER-positive breast cancer subtypes was analyzed. There were no significant differences in mean, standard deviation (SD), skewness and tumor size between Luminal A and Luminal B groups (P>0.05). The kurtosis, inhomogeneity and entropy could effectively distinguish between the two groups with statistically significant difference (P=0.001, P=0.000, and P=0.000). The area under the receiver operating characteristic (ROC) curve (AUC) of kurtosis, inhomogeneity and entropy diagnosed with malignant mass were 0.832, 0.859 and 0.891, respectively (P<0.01). In addition, the entropy was the best among the three indicators. When the entropy was ≤4.22, the sensitivity of the diagnosis Luminal B was 90.62% and the specificity was 78.95%. The texture analysis features based on DCE-MRI can help to identify ER-positive breast cancer subtypes. Entropy can be the best single texture indicator.

YAP1 positive small-cell lung cancer subtype is associated with the T-cell inflamed gene expression profile and confers good prognosis and long term survival.

9019 Background: The dominant expression of transcription factors ASCL1, NeuroD1, YAP1 or POU2F3 characteristically defines four small cell lung cancer (SCLC) subtypes (SCLC-A, SCLC-N, SCLC-Y and SCLC-P). The clinical validation and biological relevance of these emerging SCLC subtypes is currently lacking. Methods: Using the Illumina TruSeq RNA Exome Kit, we generated RNA-Seq data from 61 cases of SCLC and pulmonary carcinoid to interrogate gene expression differences in SCLC subtypes as well as in survival outliers (top and bottom decile) matched for clinically relevant prognostic factors and treatment. We also assessed YAP1 protein expression in a blinded fashion by immunohistochemistry in 130 SCLC cases. Results: We successfully classified 68% of SCLC into one of the four SCLC subtypes whereas 81.5% of carcinoids did not fit into any of these categories. GSEA for differentially expressed genes between outlier subgroups showed significant upregulation of interferon gamma and interferon alpha response genes in late survivors. Moreover, a previously validated 18-gene T-cell inflamed gene expression profile was upregulated in late survivors and in the SCLC-Y subtype. Furthermore, the SCLC-Y subtype and late survivors showed higher expression of HLA gene family and reduced expression of cancer testis antigens. The median (95%CI) OS was 14 (4.3, 28.8), 16.7 (0.9, NA), 8.1 (2, 9.7) and 20.1 (0.6, 39.5) months respectively, for SCLC-A, N, P and Y subtypes. YAP-1 protein expression was positive in 17 of 130 (13%) SCLC cases. The majority of cases with positive YAP1 expression by immunohistochemistry, 12 of 17 cases (70.6%), were limited stage SCLC at the time of original diagnosis. Conclusions: SCLC subtypes have clinical implication as predictive and prognostic biomarker. SCLC-Y subtype is enriched for T-cell inflamed phenotype and long term survival, and may predict for clinical benefit of immunotherapy.

Characterizing diversity in the tumor-immune microenvironment of distinct subclasses of gastroesophageal adenocarcinomas

Gastroesophageal adenocarcinomas (GEAs) are heterogeneous cancers where immune checkpoint inhibitors have robust efficacy in heavily inflamed microsatellite instability (MSI) or Epstein-Barr virus (EBV)-positive subtypes. Immune checkpoint inhibitor responses are markedly lower in diffuse/genome stable (GS) and chromosomal instable (CIN) GEAs. In contrast to EBV and MSI subtypes, the tumor microenvironment of CIN and GS GEAs have not been fully characterized to date, which limits our ability to improve immunotherapeutic strategies. Here we aimed to identify tumor-immune cell association across GEA subclasses using data from The Cancer Genome Atlas (N= 453 GEAs) and archival GEA resection specimen (N= 71). The Cancer Genome Atlas RNAseq data were used for computational inferences of immune cell subsets, which were correlated to tumor characteristics within and between subtypes. Archival tissues were used for more spatial immune characterization spanning immunohistochemistry and mRNA expression analyses. Our results confirmed substantial heterogeneity in the tumor microenvironment between distinct subtypes. While MSI-high and EBV+ GEAs harbored most intense T cell infiltrates, the GS group showed enrichment of CD4+ T cells, macrophages and B cells and, in ∼50% of cases, evidence for tertiary lymphoid structures. In contrast, CIN cancers possessed CD8+ T cells predominantly at the invasive margin while tumor-associated macrophages showed tumor infiltrating capacity. Relatively T cell-rich 'hot' CIN GEAs were often from Western patients, while immunological 'cold' CIN GEAs showed enrichment of MYC and cell cycle pathways, including amplification of CCNE1. These results reveal the diversity of immune phenotypes of GEA. Half of GS gastric cancers have tertiary lymphoid structures and are therefore promising candidates for immunotherapy. The majority of CIN GEAs, however, exhibit T cell exclusion and infiltrating macrophages. Associations of immune-poor CIN GEAs with MYC activity and CCNE1 amplification may enable new studies to determine precise mechanisms of immune evasion, ultimately inspiring new therapeutic modalities.

LAT1 (SLC7A5) Overexpression in Negative Her2 Group of Breast Cancer: A Potential Therapy Target.

Objective:HER2 negative carcinomas of the breast pose a challenge for treatment due to redundancies in potential drug targets and poor patient outcomes. Our aim was to investigate the role of L-type amino acid transporter – LAT1 as a potential prognosticator and a drug target. Methods:In this retrospective work, we have studied the expression of LAT1 in 145 breast cancer tissues via immunohistochemistry. Overall survival analysis was used to evaluate patient outcome in various groups of our cohort. Results:Positive LAT1 expression was found in 27 (84.4%) luminal A subtype, 27 (64.3%) luminal B/triple positive subtype, 29 (82.9%) triple negative subtype, and 24 (66.7%) HER2-only positive subtype (p=0.1). Interestingly, negative correlation was found between LAT1 and HER2; where positive expression of LAT1 was found in 56 (83.6%) cases in negative HER2 group and 51 (65.4%) cases from positive HER2 group (p=0.01). Unfortunately, we were unable to report significant survival differences when LAT1 expression was studied in the negative HER2 group. Nevertheless, five incidents of mortality (out of 55) were reported in LAT1+/HER2- group compared to none in the LAT1-/HER2- group (N=11). Conclusion:Our findings of overexpression of LAT1 in negative HER2 group suggest a role of this protein as prognosticator and drug target in a challenging therapeutic cohort.

Cytotoxic T-cell-related gene expression signature predicts improved survival in muscle-invasive urothelial bladder cancer patients after radical cystectomy and adjuvant chemotherapy

BackgroundAssessment of the immune status of muscle-invasive bladder cancer (MIBC) has previously shown to be prognostically relevant after treatment with curative intent. We conducted this study to develop a clinically...

Racial Differences in the Association Between Luminal Master Regulator Gene Expression Levels and Breast Cancer Survival.

Compared with their European American (EA) counterparts, African American (AA) women are more likely to die from breast cancer in the United States. This disparity is greatest in hormone receptor-positive subtypes. Here we uncover biological factors underlying this disparity by comparing functional expression and prognostic significance of master transcriptional regulators of luminal differentiation. Data and biospecimens from 262 AA and 293 EA patients diagnosed with breast cancer from 2001 to 2010 at a major medical center were analyzed by IHC for functional biomarkers of luminal differentiation, including estrogen receptor (ESR1) and its pioneer factors, FOXA1 and GATA3. Integrated comparison of protein levels with network-level gene expression analysis uncovered predictive correlations with race and survival. Univariate or multivariate HRs for overall survival, estimated from digital IHC scoring of nuclear antigen, show distinct differences in the magnitude and significance of these biomarkers to predict survival based on race: ESR1 [EA HR = 0.47; 95% confidence interval (CI), 0.31-0.72 and AA HR = 0.77; 95% CI, 0.48-1.18]; FOXA1 (EA HR = 0.38; 95% CI, 0.23-0.63 and AA HR = 0.53; 95% CI, 0.31-0.88), and GATA3 (EA HR = 0.36; 95% CI, 0.23-0.56; AA HR = 0.57; CI, 0.56-1.4). In addition, we identify genes in the downstream regulons of these biomarkers highly correlated with race and survival. Even within clinically homogeneous tumor groups, regulatory networks that drive mammary luminal differentiation reveal race-specific differences in their association with clinical outcome. Understanding these biomarkers and their downstream regulons will elucidate the intrinsic mechanisms that drive racial disparities in breast cancer survival.

Lipidomic study of cell lines reveals differences between breast cancer subtypes.

Breast cancer (BC) is the most prevalent type of cancer in women in western countries. BC mortality has not declined despite early detection by screening, indicating the need for better informed treatment decisions. Therefore, a novel noninvasive diagnostic tool for BC would give the opportunity of subtype-specific treatment and improved prospects for the patients. Heterogeneity of BC tumor subtypes is reflected in the expression levels of enzymes in lipid metabolism. The aim of the study was to investigate whether the subtype defined by the transcriptome is reflected in the lipidome of BC cell lines. A liquid chromatography mass spectrometry (LC-MS) platform was applied to analyze the lipidome of six cell lines derived from human BC cell lines representing different BC subtypes. We identified an increased abundance of triacylglycerols (TG) ≥ C-48 with moderate or multiple unsaturation in fatty acyl chains and down-regulated ether-phosphatidylethanolamines (PE) (C-34 to C-38) in cell lines representing estrogen receptor and progesterone receptor positive tumor subtypes. In a cell line representing HER2-overexpressing tumor subtype an elevated expression of TG (≤ C-46), phosphatidylcholines (PC) and PE containing short-chained (≤ C-16) saturated or monounsaturated fatty acids were observed. Increased abundance of PC ≥ C-40 was found in cell lines of triple negative BC subtype. In addition, differences were detected in lipidomes within these previously defined subtypes. We conclude that subtypes defined by the transcriptome are indeed reflected in differences in the lipidome and, furthermore, potentially biologically relevant differences may exist within these defined subtypes.

Whole-lesion histogram analysis of apparent diffusion coefficient for the prediction of pathological complete response to neoadjuvant chemotherapy in different subtypes of breast cancer

Objective To investigate the value of whole-lesion histogram parameters of apparent diffusion coefficient (ADC) in evaluating and predicting the pathological complete response(PCR) to neoadjuvant chemotherapy (NAC) in different subtypes of breast cancer. Methods This retrospective study included 117 patients with breast cancer who underwent MRI examination before NAC prior to surgery from January 2016 to December 2017 in the First Affiliated Hospital of Nanjing University. All cases were divided into Luminal B, HER2 positive (n=21) and triple negative (n=26) groups. The surgical pathology after chemotherapy was evaluated by Miller-Payne (M-P) system and the patients were divided into PCR group and non-PCR (nPCR) group. Firevoxel software was used to generate the whole-lesion ADC histogram. The parameters included mean (ADCmean), skewness, kurtosis, the minimum (ADCmin), the maximum (ADCmax), 10th percentile(ADC10%), 50th percentile (ADC50%) and 90th percentile (ADC90%). The two independent samples t test or Mann-Whitney U test was used to compare the differences between PCR and nPCR groups in each subtype. The diagnostic performance of statistically different ADC parameters for predicting PCR was evaluated by receiver operating characteristic (ROC) curve. Results Kurtosis was significantly higher in PCR group than that in nPCR group in HER2 positive subtype (P=0.039). It achieved an area under the curve (AUC) of 0.813 with sensitivity of 100% and specificity of 68.7% at the optimal cutoff value (1.861) for differentiating PCR from nPCR cases. In triple negative subtype, ADCmean and ADC50% were smaller in PCR group than those in nPCR group (P=0.028,0.013). They achieved AUCs of 0.800, 0.842, respectively. When ADCmean of 1.030×10-3 mm2/s and ADC50% of 0.976×10-3 mm2/s were used as cutoff value to differentiate PCR from nPCR, the sensitivities were 75.0%, 80.0% and the specificities were 83.3%, 83.3%, respectively. Conclusion Kurtosis can predict post-NAC PCR in patients with HER2 positive breast cancer, while ADC50% has a high value in predicting post NAC PCR of triple negative breast cancer patients. Key words: Breast neoplasms; Molecular subtype; Neoadjuvant chemotherapy; Magnetic resonance imaging

PIK3CA mutation profiling in Vietnamese patients with breast cancer

Activating mutations in phosphoinositide-3-kinase, catalytic, alpha polypeptide (PIK3CA) gene frequently occur in breast cancer, and are important for the survival of patients with breast cancer. Although PIK3CA mutations have been studied widely in many countries, the relationships between PIK3CA mutations and clinicopathological features are still inconsistent. Moreover, information about PIK3CA mutations in Vietnamese breast cancer has not been investigated. In the present study, we identified PIK3CA mutations in 27.2% of 162 human breast tumors, including 31 exon 9 mutations and 13 exon 20 mutations. In addition, PIK3CA mutations were associated with larger tumor size and low Ki67 level. Moreover, the distribution of PIK3CA mutations in the breast cancer subtypes was analyzed. The PIK3CA mutations was not equally distributed among different intrinsic subtypes, in which the rate of mutations in hormone (HR)-positive subtypes was somewhat higher than that in human epidermal growth factor receptor-2 (HER2)-positive subtypes. Overall, this study provides more information about PIK3CA mutations and suggests prognostic and predictive values for breast cancer management in Vietnamese population.

Construction of differentially expressed Her-2 related lncRNA-mRNA-miRNA ceRNA network in Her-2 positive breast cancer.

BackgroundHer-2 positive subtype breast cancer is characterized as Her-2 gene amplification with poor survival and increased invasiveness accounting for 20–30% of invasive infiltrated breast cancer. A lncRNA-miRNA-mRNA competing endogenous RNA (ceRNA) network is constructed to detect Her-2 specific RNAs in the development and progression of HER-2 positive breast cancer which may overcoming the anti-HER-2 therapy resistance of breast cancer cells.MethodsOne thousand one hundred and nine breast cancer samples obtained from The Cancer Genome Atlas (TCGA) database were classified into two cohorts including ER+/PR+ (n=461) and ER-/PR- breast cancer (n=152). Differently expressed mRNAs, lncRNAs and miRNAs were screened in ER+/PR+ and ER-/PR- breast cancer cohorts, respectively. lncRNA-miRNA interactions were preformed to predicted and verified by miRcode. miRNA-mRNA interactions were selected to predict targeted mRNAs of miRNAs by miRanda, Targetscan and miRTarBase.ResultslncRNA-miRNA-mRNA ceRNA network was constructed by retained lncRNAs, miRNAs and mRNAs. Fifteen DEmiRNAs, 129 DElncRNAs and 269 DEmRNAs were retained in ER+/PR+ cohort after intersection with DEmiRNAs, DElncRNAs and DEmRNAs between breast cancer and normal tissues. Six hundred and ninety-three DEmRNAs, 25 DEmiRNAs and 364 DElncRNAs were retained in ER-/PR- cohort. ceRNA network in ER+/PR+ breast cancer cohort was constructed of the interactions of 4 DElncRNA–DEmiRNA pairs and 2 DEmiRNA–DEmRNA pairs included 4 DElncRNAs, 1 DEmiRNAs, and 2 DEmRNAs. ceRNA network in ER-/PR- breast cancer cohort was constructed of the interactions of 24 DElncRNA–DEmiRNA pairs and 1 DEmiRNA–DEmRNA pairs included 19 DElncRNAs, 4 DEmiRNAs, and 1 DEmRNA. MIR7-3HG- hsa-mir-204-NTRK2 axis was identified in both ER+/PR+ and ER-/PR- cohort in our study.ConclusionsBased on the ceRNA hypothesis, a potential Her-2 related regulatory ceRNA networks are constructed which may provide novel insights into the mechanism underlying the biological processes of Her-2 positive breast cancer.

Estimating breast tissue-specific DNA methylation age using next-generation sequencing data

BackgroundDNA methylation (DNAm) age has been widely accepted as an epigenetic biomarker for biological aging. Emerging evidence suggests that DNAm age can be tissue-specific and female breast tissue ages faster than other parts of the body. The Horvath clock, which estimates DNAm age across multiple tissues, has been shown to be poorly calibrated in breast issue. We aim to develop a model to estimate breast tissue-specific DNAm age.MethodsGenome-wide DNA methylation sequencing data were generated for 459 normal, 107 tumor, and 45 paired adjacent-normal breast tissue samples. We determined a novel set of 286 breast tissue-specific clock CpGs using penalized linear regression and developed a model to estimate breast tissue-specific DNAm age. The model was applied to estimate breast tissue-specific DNAm age in different breast tissue types and in tumors with distinct clinical characteristics to investigate cancer-related aging effects.ResultsOur estimated breast tissue-specific DNAm age was highly correlated with chronological age (r = 0.88; p = 2.9 × 10−31) in normal breast tissue. Breast tumor tissue samples exhibited a positive epigenetic age acceleration, where DNAm age was on average 7 years older than respective chronological age (p = 1.8 × 10−8). In age-matched analyses, tumor breast tissue appeared 12 and 13 years older in DNAm age than adjacent-normal and normal breast tissue (p = 4.0 × 10−6 and 1.0 × 10−6, respectively). Both HER2+ and hormone-receptor positive subtypes demonstrated significant acceleration in DNAm ages (p = 0.04 and 3.8 × 10−6, respectively), while no apparent DNAm age acceleration was observed for triple-negative breast tumors. We observed a non-linear pattern of epigenetic age acceleration with breast tumor grade. In addition, early-staged tumors showed a positive epigenetic age acceleration (p = 0.003) while late-staged tumors exhibited a non-significant negative epigenetic age acceleration (p = 0.10).ConclusionsThe intended applications for this model are wide-spread and have been shown to provide biologically meaningful results for cancer-related aging effects in breast tumor tissue. Future studies are warranted to explore whether breast tissue-specific epigenetic age acceleration is predictive of breast cancer development, treatment response, and survival as well as the clinical utility of whether this model can be extended to blood samples.

Neoadjuvant chemotherapy in non-metastatic breast cancer: a study on practice trends in a regional cancer treatment service.

Neoadjuvant chemotherapy (NACT) is increasingly used for managing locally advanced and high risk non-metastatic breast cancer. To describe trends in NACT use, assess compliance to best practice recommendations and determine treatment response rates in a regional cancer treatment service. In this retrospective cross- sectional study, electronic records of patients who underwent NACT in centres covered by the MidCentral Regional Cancer Treatment Service in 2013 and 2017 were reviewed. Data pertaining to patient demographics, disease status, compliance to best practice recommendations and treatment outcomes were extracted and analysed. Of a total of 502 referrals for non-metastatic breast cancer, 34 underwent NACT with the estimated NACT rate rising from 3.85% (2013) to 9.92% (2017). Compliance to practice recommendations improved in all domains (pre-treatment tumour and axillary evaluation, marker placement, multidisciplinary discussion). Overall, NACT was well tolerated with only three patients experiencing treatment limiting toxicity. Response rates mirror published data (complete response: 29.4%, partial: 61.8%) with higher responses registered in HER2 positive and triple negative subtypes. Discordance between radiological and pathological response was 28%, with imaging overestimating response in five out of seven cases. Of the 11 (32%) patients who initially underwent breast conserving surgery, six required a second surgery. NACT is increasingly used in the Regional Cancer Treatment Service, with improving compliance to practice recommendations. These results are reassuring and can be used to help patients develop a realistic expectation towards NACT.

Clinico-pathological co-relation using various immuno-histochemistry markers likeER, PR, HER-2 NEU, CK5/6, EGFR, KI-67 in carcinoma breast

In India, for the year 2012, 144,937 women were newly detected with breast cancer and 70,218 women died of it. For every 2 women newly diagnosed with breast cancer, one lady is dying of it. The aim of this study is to evaluate clinical parameters and pathological findings including various Immuno-histochemistry (IHC) markers like ER, PR, HER-2 NEU, CK5/6, EGFR, Ki-67 in cases of carcinoma breast and classify them into molecular classification based on IHC markers and try to correlate them clinically. This prospective, observational study was carried out in 56 patients with early carcinoma breast (stage-I and stage-II) and IHC evaluation for various markers was done. Data was analysed by using Molecular Classification, divide them into estrogen positive (luminal HER-2, luminal A and luminal B) and estrogen negative (Triple negative or basal cell type, HER-2Neu type and normal breast like phenotype) subtypes. We had correlated this data with parameters like age of the patient, clinical and pathological staging of the breast carcinoma, presence or absence of nodes and presence or absence of other IHC parameters. We used ANOVA-F test to catagories variables and measure the test of significance. On IHC in Her-2 neu equivocal cases (patients who had two “++” positive points), we performed FISH test. Out of these 17 equivocal cases, only 3 were positive, 10 were negative and 4 patients did not underwent this test due to several reasons. Finally, Ki-67 value is significantly high in triple negative and Luminal-B patients. NPI is also having low ‘P’value, although not reaching the level of significance. Types of breast carcinoma, which look histologically similar behaves differently in their clinical presentation and in prognosis.In our study only Ki-67 was correlated with poor prognostic subtype of molecular classification but no any poor risk of clinical or histological parameter was correlated significantly with bad prognostic subtype of molecular classification as Luminal-B or triple negative type. We can say that this molecular classification is different in terms of prognosis in patients with similar looking clinical and histological parameters.

Abstract P6-06-13: Racial/ethnic differences in tumor infiltrating lymphocytes

Abstract Background/Rationale: There have been increasing studies regarding the morphologic evaluation of tumor-infiltrating lymphocytes (TILs) and the tumor microenvironment. In breast cancer, TILs are more abundant in stroma (sTILs) compared to intratumoral (iTILs) areas and iTILs have been an unreliable marker due to difficulty in standardizing their evaluation. The International Immuno-Oncology Working Group published recommendations to guide the accurate assessment of TILs. TILs have relevance in the setting of neoadjuvant therapy (NAT), primarily in triple negative and HER2 positive breast cancers. Studies report that sTIL infiltration is predictive of response to NAT in these two breast cancer subtypes, but not in the estrogen positive breast cancer subtype. The presence of TILs is a good prognostic indicator and correlates with axillary lymph node negativity, lower histological grade and improved recurrence-free survival. Therefore, evaluating TILs prior to NAT can provide important predictive and prognostic information for clinicians. There are insufficient data exploring differences in sTILs among different racial/ethnic groups. Previous data showed Asians had more sTILs compared to White and African American patients, consistent with data from Asia that found substantial sTILs in their respective patient populations. Our diverse population has known racial/ethnic disparities, where Pacific Islanders have a higher mortality compared to Asian and White patients. Evaluating sTILs within this population may reveal tumor biology differences that could contribute to health disparities. Research objectives: To determine the stromal tumor infiltrating lymphocytes on core biopsy specimens and correlate them with clinical characteristics: ethnicity, age, BMI and stage To determine differences in pathologic complete response (pCR) with ethnicity, age, BMI, neoadjuvant therapy and stage Table 1. % Stromal tumor infiltrating lymphocytes (sTILs) compared to age, BMI, ethnicity, stage and pCR adjusted for breast cancer subtypeVariableValueNMeanSDPAge30838.615.140927.123.0.30501834.626.6.67601431.121.3.4570+826.822.8.30BMI&amp;lt; 252825.918.825-301638.521.9.07&amp;gt; 301337.228.0.12EthnicityAsian1636.724.2.01Pacific Islander2038.021.3.004White1818.515.9Other348.130.6.02Stage1A818.48.01B933.425.5.192A1631.923.0.172B938.620.6.113A630.823.2.313B937.225.8.10pCRno3233.019.8yes2530.825.6.72 Table 2. Pathologic complete tesponse (pCR) compared to age, BMI, neoadjuvant therapy, ethnicity and stage adjusted for breast cancer subtypeVariableValueNMeanSDPAge3080.5500.5354090.4060.527.5650180.4450.511.6360140.3030.469.2770+80.5860.518.89BMI&amp;lt; 25280.5260.50825-30160.2730.447.11&amp;gt; 30130.4530.519.65Neoadjuvant TherapyACT140.2680.469.88TC110.2370.505TCHP160.7480.479.02THP120.4280.515.41Letrozole40.3850.500.62EthnicityAsian160.4720.512.77Pacific Islander200.3890.503.85White180.4200.511other30.7060.577.36Stage1A80.2410.3541B90.4270.527.412A160.7240.479.022B90.0320.441.403A60.8220.516.023B90.2680.500.90 Results: We evaluated 57 neoadjuvant breast cancer cases for sTILs according to the International Immuno-Oncology Working Group recommendations and reviewed their clinical characteristics. We found significant % sTILs differences in ethnicity, specifically Asian (35%), Pacific Islander (38.5%) and Other (46%) category compared to White (19%) Table 1. There were no differences found in sTILs according to age, BMI or stage. We also found no significant pCR differences according to age, BMI or ethnicity, however for certain stages and neoadjuvant therapy, there is an increased rate of pCR, although these are a small number of cases, no conclusions can be made, Table 2. Future studies to evaluate the significance of sTILs in different ethnicities may be informative to delineate possible mechanisms contributing to known disparities they experience. Citation Format: Jami Fukui, Alana Taniguchi, Madison Meister, Ian Pagano, Jeffrey Killeen. Racial/ethnic differences in tumor infiltrating lymphocytes [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 P6-06-13.

Abstract P1-10-23: Prediction of pathological complete response (pCR) upon neoadjuvant chemotherapy by MammaTyper® pCR score

Abstract Background: Neoadjuvant chemotherapy (NACT) (+/- anti HER2) is the preferred therapy for triple negative and HER2 positive subtype breast tumors. In Luminal subtype breast tumors, NACT is used to downstage large inoperable tumors and enable breast conserving therapy. However, this subgroup of patients may be overtreated as many Luminal subtype tumors do not respond to NACT due to the low rate of proliferating tumor cells. Herein, we tested the MammaTyper® pCR score as a predictor of therapy success measured as pCR in a retrospective analysis of archived samples from a single center. Methods: 85 FFPE biopsy samples from the years 2012-2018 were sourced from the archive. Samples with &amp;gt;20% tumor cell content were subjected to RNA extraction. Relative mRNA expression levels of ERBB2, ESR1, PGR and MKI67 were determined by RT-qPCR using the CE-IVD MammaTyper® kit. The continuous expression values were integrated into a prediction score (pCR-score) using a pre-defined algorithm and cutoff. The association of continuous and binary pCR-score results with pCR (defined as ypT0/Tis) and partial response was analyzed. Results: Marker positivity rates of the 75 samples included in the final analysis were 62.7% for ER, 53.3% for PR, 40.0% for HER2 and 94.7% for Ki67 (≥20% positive cells). 42.7% of patients were pre-menopausal and all samples except one were grade 3. pCR rates over all samples and in hormone receptor (HR) positive/HER2 negative patients only were 48.0% and 20.0%, respectively. Binary pCR-score result was significantly associated with pCR over all patients (Sensitivity: 88.9%, Specificity: 51.3%, PPV: 62.8%, NPV: 83.3%) and also in HR positive/HER2 negative patients only (Sensitivity: 83.3%, Specificity: 70.8%, PPV: 41.7%, NPV: 94.4%). ROC analysis revealed a good association of the continuous pCR-score with achievement pCR over all patients (AUC=0.756) and in the subgroup of HR+/HER2- patients (AUC=0.774). pCR rates according to St Gallen surrogate subtype definition were similar for IHC and RT-qPCR defined subtypes in Triple Negative (80.0% and 78.6% respectively) and in the HER2+ non luminal subtype (75.0%, and 70.0% respectively). In tumors with incomplete response the continuous pCR-score was significantly associated with residual tumor size (Spearman rs: -0.477 p-value: 0.0021) and %-decrease of tumor size (Spearman rs: 0.388, p-value: 0.0147). Conclusion: The MammaTyper® pCR score may serve as a standardized tool to predict response to NACT based on a pre-treatment biopsy. For patients with inoperable Luminal tumors and low predicted probability of pCR, neo-adjuvant aromatase inhibitor alone or combined with the new generation of TKIs such as CDK4/6 inhibitors or Pi3KCa/mTOR inhibitors may be an alternative for downstaging of tumors. Citation Format: Silvana Saracchini, Anna Bassini, Wally Marus, Serena Corsetti, Ilaria Specogna, Manuela Bertola, Elvia Micheli, Ralph M Wirtz, Mark Laible, Ugur Sahin, Daniele Generali, Sandro Sulfaro. Prediction of pathological complete response (pCR) upon neoadjuvant chemotherapy by MammaTyper® pCR score [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 P1-10-23.

Abstract P1-10-08: Assessing immune biomarkers of response to anthracyclines in breast cancer

Abstract Background: Pathologists have long recognized that the interaction between immune and tumour cells is critical in the development and progression of breast cancer. Studies have demonstrated the presence of tumour-infiltrating lymphocytes (TILs) correlates with improved clinical outcome in breast cancer especially in the triple negative and HER-2 positive subtypes. TILs predict for improved response to certain therapies including chemotherapy and trastuzumab. The predictive value of TILs in ER positive tumours is less clear. It has been demonstrated the higher presence of the immune microenvironment is associated with a better prognosis and as a result a higher likelihood of benefit from chemotherapy and possibly from immunotherapy, whereas cold immune microenvironment carries greater risk of relapse and lower benefit from chemotherapy and possibly immuno-therapies. In this study, we evaluated whether TILs could be used to predict chemotherapy response and characterize the pre-existing tumour microenvironment (TME) using NanoString’s GeoMx Digital Spatial Profiling (DSP) platform. Methods: We assessed haematoxylin and eosin stained slides from the phase III BR9601 adjuvant breast cancer trial using software used in the international ring study 2 for standardized evaluation of TILs integrated in VMscope slide explorer. Evaluation of stromal TILs was based on international guidelines. NanoString’s DSP platform was used to analysis 256 patient samples from the BR9601 clinical trial. For analysis, region of interest were selected and compared for the TME (CD45+ve) and tumour rich (pan cytokeratin) in tissue microarrays. A panel of 56-antibodies were analysed in each ROI. Results: The mean TIL score in this cohort of patients was 15.58% (ranging from 0 to 66.67%). The presence of higher levels of TILs was significantly associated with ER negativity (p&amp;lt;0.001), high grade (p=0.01) and increased lymph nodal involvement (p=0.002). In multivariate analysis, patients whose tumours had medium/high levels of TILS expression had better DRFS (HR: 0.49, 95%CI 0.24-1.02, p=0.057) when treated with E-CMF than those treated with CMF alone. Highest levels of TILs were found in Basal and HER2-like tumours. A T-cell score was generated using the average expression of CD3, CD4 and CD8. The T-cell score was examined in both the tumour and TME. Using the T cell score it was apparent that the cohort had a range of immune “hot” and immune “cold” tumours. It was demonstrated that immune “hot” TME doesn’t not always correlate with immune “hot” tumour expression. Proteins that were most associated with T-cell exclusion (p&amp;lt;0.01) in the TME were Fibronectin, B7-H3, PTEN, ER-α, TGFB1, FAPα and CD34. This would indicate that these proteins are causal inhibitors of T-cell invasion. Conclusion: In conclusion, this study highlights the significance of assessing the entire tumour since TILs, tumour and stromal cells collectively engage in a complex interplay that contributes to disease development and progression. NanoString’s GeoMx DSP is a promising technology for multiplexed analysis. TILs, whether measured using automated software, or estimation by protein profiling, are predictive of chemotherapy benefit. Citation Format: Melanie Spears, Carsten Denkert, Sonia L Villagas, Nicola Lyttle, Linda Liao, Mary Anne Quintayo, Christopher J Twelves. Assessing immune biomarkers of response to anthracyclines in breast cancer [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 P1-10-08.

Abstract P2-08-05: Early breast cancer in oldest population: Screening, tumor characteristics and treatment choices in octogenarian women

Abstract Introduction: Breast cancer risk increases with age and about a third of female breast cancers are diagnosed in patients (pts) aged older than 70. Although we could expect that elderly pts will be treated with similar intensive treatment as applied in younger age groups, octogenarian are frequently undertreated, even after adjustment for confounding factors, such as co-morbidities, social support and functional status. Methods: This retrospective study investigated women referred for surgical evaluation of biopsy-proven primary early-stage invasive breast cancer from 2008 - 2018. From medical record, we obtained standard clinic-pathologic data, co-morbidities, treatment (surgery, radiotherapy, chemotherapy, and endocrine therapy), and Eastern Cooperative Oncology Group (ECOG) functional status affecting clinical decision. Results: The study identified 112 eligible women between 80- 90 years, median 82 years (rIQ 80-84); diagnosed by screening 27 pts (24%), palpable mass 53 pts (47%) and unknown 32 pts (29%). Stage I 45 (40%), II 34 (30%), III 17 (15%), unknown 16 (14%). ECOG 0: 60 pts (53%), 1: 19 pts (18.5%), 2: 16 pts (14%). Forty pts (35%) had 1 co-morbidity, 2 pts (15%) 2 and 10 pts (8%) 3 or more. Only 6 pts (5%) were referred to geriatric consultation. Tumor characteristics: ductal 62 (56%) and lobular 19 (18%), histological grade 1 or 2 68 (60%); Luminal tumors 89 (79%), Her-2 positive 8 (7%) and triple negative 9 (8%), unknown 6 (4%). Lumpectomy was the most frequent local treatment in 77 pts (69%), with sentinel lymph node biopsy 47 pts (63%), with lymphadenectomy 13 pts (18%) and 17 pts (19%) did not receive axillary treatment. In 22 pts (19%) mastectomy was performed, 14 with lymphadenectomy. Whole breast radiotherapy was indicated in 39 pts (52%) with lumpectomy (38 pts with lumpectomy did not receive radiation therapy). Only 5 from 9 pts with 4 or more axillary positive nodes received radiotherapy in lymph node areas. In HER-2 positive subtype, 1 pt was treated with paclitaxel/trastuzumab, 3 pts received trastuzumab and 4 pts did not receive any treatment. Luminal subtype (Her-2 negative) with positive axillary were 21 pts (18%), solely 9 received adjuvant chemotherapy. Only six pts with triple negative tumors received adjuvant chemotherapy. Adjuvant endocrine therapy was indicated in 78 patients: 40 pts with tamoxifen and 38 pts with aromatase inhibitors. Thirty eight pts had adjuvant chemotherapy criteria but 22 pts did not receive cytotoxic drugs. The majority of these treatments not indicated were based on the decision of the attending physician. Only 3 pts with high risk characteristics were evaluated on geriatric consultation. The median follow-up was 27 months. Conclusions: The most frequent breast cancer subtype in our population was luminal, and there was a lower prevalence of Her-2 positive and triple negative tumors in comparison with pts between 50-70 years. Despite the low number of cases detected by screening, most pts were Stage I and II, probably due to our population conditions (high income women, private institution). Conservative surgery was the most frequent local treatment, nevertheless the indication of whole breast radiotherapy was lower than expected. Moreover systemic chemotherapy in high-risk patients (Luminal with axillary involvement, Her-2 and triple negative) were less than recommended by international treatment guidelines and, in most cases, decision was made by the attending physician without previous geriatric consultation. We suggest multidisciplinary management of this subgroup of elderly patients, perform oncogeriatric consultation and use objective evaluation parameters (such as the G8 scale) Citation Format: Sergio G Rivero, Alexis Ostinelli, Maria V Costanzo, Adrián Nervo, Jorge Nadal, Federico Coló, Carlos M Loza, Mora Amat, Verónica Fabiano, Roberto Gavazzi, Reinaldo Chacón. Early breast cancer in oldest population: Screening, tumor characteristics and treatment choices in octogenarian women [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 P2-08-05.

Does function predict persistence? Nonsuicidal self-injury among adolescents during and after hospitalization

Nonsuicidal self-injury (NSSI) is a prevalent, concerning behavior among adolescents. Importantly, NSSI can serve a variety of functions. Some adolescents engage in NSSI to fulfill automatic or self-oriented functions (e.g., cutting to avoid internal negative states), whereas others engage in NSSI to serve social functions (e.g., cutting to communicate with others). This study tests whether self-reported reasons for engaging in NSSI, hereafter referred to as NSSI functions, predict NSSI thoughts and behaviors during and after hospitalization among adolescent psychiatric inpatients. Endorsement of both automatic and social NSSI functions, as well as positive and negative reinforcement subtypes, was assessed at hospital admission. Results showed that endorsement of overall automatic function predicted which adolescents engaged in NSSI behavior during hospitalization. Moreover, automatic and social functions showed distinct predictive patterns, such that automatic functions corresponded to greater likelihood of NSSI-related thoughts and behaviors whereas social functions mainly corresponded to reduced likelihood of NSSI-related outcomes. Of note, NSSI functions were less predictive of NSSI-related outcomes after hospital discharge. These findings suggest that identifying adolescent inpatients’ reasons for NSSI engagement may meaningfully distinguish those at higher risk (and those at lower risk) of NSSI persistence during their hospital stay.

Pathologic assessment of tumor-associated macrophages and their histologic localization in invasive breast carcinoma

BackgroundTumor-associated macrophages (TAMs) are important in regulating cross-talk between tumor cells and tumor microenvironment. TAMs are involved in multiple steps of tumor progression and invasion. This study aimed to compare CD163 expression with the widely used CD68 pan-macrophage marker in invasive breast carcinoma. Furthermore, it focused on assessing the significance of TAMs localization in relation to clinicopathological parameters.ResultsCD68 and CD163 immunohistochemical expressions within TAMs infiltrating both tumor nest (TN) and tumor stroma (TS) were evaluated in 60 specimens with invasive breast carcinoma. High CD68-positive stromal TAMs was significantly related to larger tumor, nodal metastasis and vascular invasion (p = 0.003, 0.037, 0.032, respectively), whereas high CD163-positive stromal TAMs was significantly related to larger tumors, nodal metastasis, stage III tumors, vascular invasion, estrogen receptor (ER) negativity, and triple-negative subtype (p = 0.023, < 0.001, 0.001, 0.022, 0.002, 0.017, respectively). On multivariate analysis, high CD68-positive TAMs infiltrating TS was significantly associated with larger tumor and positive nodal metastasis (p = 0.006 and 0.016, respectively), whereas high CD163 TAMs density within TS was significantly associated with positive vascular invasion, nodal metastasis, and molecular subtypes (p = 0.003, 0.001, and 0.009, respectively).ConclusionTAMs within tumor stroma and tumor nest have different levels of association with poor prognostic parameters. So, it is of great importance to consider the histologic localization of TAMs in addition to the degree of TAMs infiltration.

An observational study of clinical subtypes in treatment-resistant schizophrenia and its relation with cognitive insight

Background: Schizophrenia is a chronic disorder, and one-third of patients with schizophrenia can develop treatment resistance. Factors such as male gender, early age of onset, positive family history, and low level of social functioning are associated with refractoriness to treatment. However, very few Indian studies have tried to establish the predictors of outcome for schizophrenia. Aims: The aim of the study was to identify the subtypes of treatment-resistant schizophrenia (TRS) and its relation with sociodemographic profile, illness-related variables, and cognitive insight. Materials and Methods: The study was conducted after permission from the institutional review board. Fifty consecutive patients of TRS (treated with two antipsychotics from different classes with adequate doses for 6 weeks each and currently satisfying the Diagnostic and Statistical Manual of Mental Disorders 4th Edition, Text Revision criteria for schizophrenia) were included in the study. Patients were administered a semi-structured questionnaire to obtain details about sociodemographic status, and the Positive and Negative Syndrome Scale was administered to identify the subtypes of schizophrenia. Becks Cognitive Insight Scale was used to assess cognitive insight. Results: Almost half of the sample (46%) belonged to the positive subtype followed by negative subtype. There was a statistically significant difference in the age of onset, educational qualification, and suicidal ideation between the positive and negative subtypes. There was a significant association of cognitive insight between TRS subtypes, and it suggested that patients with positive subtype had poorer insight than those with negative subtype. Conclusions: It is important to identify the subtype of schizophrenia and the risk factors for treatment resistance. Early recognition and intervention with respect to these risk factors can improve the long-term course of TRS.