Non-pathological Complete Response Research Articles

Targeting SGK1 enhances the efficacy of radiotherapy in locally advanced rectal cancer

Radiotherapy (RT) is a key component of neoadjuvant chemoradiotherapy to treat locally advanced rectal cancer (LARC). However, the therapeutic effect is limited due to radioresistance. Investigating the biomarkers of radioresistance might assist in the development of more effective therapeutic strategies for LARC.In this study, we investigated the different gene expressions in tumor samples from 110 patients using transcriptome analysis and immunohistochemistry (IHC), and identified serum- and glucocorticoid-regulated kinase 1 (SGK1) as a modulator of LARC radioresistance. We evaluated the impact of genetic and pharmacologic inhibition of the gene associated with radioresistance in vitro and in vivo. We found that the expression of SGK1 was upregulated in non-pathological complete response (non-pCR) patients. A high SGK1 expression was associated with radioresistance, whereas the genetic or pharmacologic inhibition of SGK1 expression reduced the radioresistance. We found that activate transcription factor 3 (ATF3) is a regulator of SGK1 in radioresistance.In conclusion, our findings indicate that SGK1 is a key player in LARC radioresistance, and drives radioresistance in an ATF3 dependent manner, which provides insights for future radio-sensitizer design.

Comparing the Outcome of Boost Dose of Intraoperative Radiotherapy with Electron (IOERT) and Low-kV X-Ray (IOXRT) and External Beam Radiotherapy (EBRT) in Breast Cancer After Neoadjuvant Chemotherapy

Background: In the large breast tumors or locally-advanced breast cancers, breast conserving surgery (BCS) after neoadjuvant chemotherapy (NACT) had an acceptable local control, but greater risk of recurrence. Adding boost dose radiation to whole breast radiotherapy is involved with a reduced risk of recurrence. Boost radiotherapy can be delivered in 3 methods, including (1) external beam radiotherapy (EBRT), (2) intraoperative radiotherapy with electron (IOERT), and (3) intraoperative radiotherapy with low-kV X-ray (IOXRT). Objectives: This study compared the outcomes of these 3 methods with each other. Methods: Within 60 months, 217 unselected breast cancer patients in Cancer Research Center of Shahid Beheshti were under treatment with BCS after NACT. They received boost dose radiation in 3 groups; 115 patients in the EBRT group, 39 patients IOXRT group, and 63 patients in the IOERT group. All of them received WBRT after surgery. Results: The patients had large tumors or stage 3 breast cancer. Local recurrences were 1 (2.5%) in IOXRT, 2 (3.2%) in IOERT, and 1 (0.9%) in EBRT groups. Systemic recurrences were 4 (10.3%) in IOXRT, 10 (15.9%) in IOERT, and 16 (13.9%) in EBRT groups. Deaths were 3 (7.7%) in IOXRT, 2 (3.2%) in IOERT, and 10 (6.9%) in EBRT groups. Patients with any events were 4 (10.3%) in IOXRT, 11 (17.5%) in IOERT, and 33 (15.2%) in the EBRT group. Death due to distant metastases was lower in IOERT group, but it was not significant. No significant difference was observed in disease-free survival (DFS) among 3 groups. IOXRT group had non-significant, lower events, and better DFS. Especially, in non-PCR (non-pathologic complete response) patients, multivariate COX analysis showed better outcome (DFS) in IOXRT group (HR = 0.50), although it was not significant (P = 0.53). Conclusions: Intraoperative radiotherapy (IORT or IOXRT) as tumor bed boost during BCS after NACT had at least non-inferiority compared with EBRT. In non-PCR patient, IOXRT group had non-significant better outcomes (DFS).

The Effect of the Leptin and Leptin Receptor Expression on the Efficacy of Neoadjuvant Chemotherapy in Breast Cancer.

BackgroundThe purpose of the present study was to evaluate the effect of leptin and leptin receptor (LEPR) expression on the efficacy of neoadjuvant chemotherapy in breast cancer.Material/MethodsThere were 325 breast cancer patients with complete data enrolled in this study. Patients were categorized into 3 groups: pathological complete response group, non-pathological complete response group, and progressive disease group. Immunohistochemistry was performed to determine leptin and its receptor LEPR expression levels that were compared among the 3 groups.ResultsCompared with the non-pathological complete response group, patients in the pathological complete response group had increased leptin and LEPR expression, although the difference was not statistically significant (P=0.194, P=0.110). In addition, the expression of leptin and LEPR in the pathological complete response group was also higher than that in the progressive disease group, and the difference of LEPR expression was statistically significant (P=0.008) while the leptin expression was not (P=0.065). There were more HER2+ breast cancer patients in the pathological complete response group categorized into strong positive, and positive expression of leptin and LEPR compared with the progressive disease group (P<0.05). There were significant differences of leptin and LEPR expression among breast cancer patients under different molecular subtypes HER2+, HR+, and triple negative, in which the triple negative patients had the highest expression of leptin and LEPR. In addition, patients in the progressive disease group had high and low expression of leptin and LEPR: 13.25% versus 11.32% and 13.1% versus 10.42% respectively.ConclusionsOverexpression of leptin and LEPR improved the therapeutic efficacy of neoadjuvant chemotherapy for patients with breast cancer, especially for those with HER2+ subtype. Overexpression of leptin and LEPR was distinct among the different molecular subtypes of breast cancer, suggesting a certain predictive value for breast cancer prognosis.

Early Prediction of Breast Cancer Therapy Response using Multiresolution Fractal Analysis of DCE-MRI Parametric Maps.

We aimed to determine whether multiresolution fractal analysis of voxel-based dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) parametric maps can provide early prediction of breast cancer response to neoadjuvant chemotherapy (NACT). In total, 55 patients underwent 4 DCE-MRI examinations before, during, and after NACT. The shutter-speed model was used to analyze the DCE-MRI data and generate parametric maps within the tumor region of interest. The proposed multiresolution fractal method and the more conventional methods of single-resolution fractal, gray-level co-occurrence matrix, and run-length matrix were used to extract features from the parametric maps. Only the data obtained before and after the first NACT cycle were used to evaluate early prediction of response. With a training (N = 40) and testing (N = 15) data set, support vector machine was used to assess the predictive abilities of the features in classification of pathologic complete response versus non-pathologic complete response. Generally the multiresolution fractal features from individual maps and the concatenated features from all parametric maps showed better predictive performances than conventional features, with receiver operating curve area under the curve (AUC) values of 0.91 (all parameters) and 0.80 (Ktrans), in the training and testing sets, respectively. The differences in AUC were statistically significant (P < .05) for several parametric maps. Thus, multiresolution analysis that decomposes the texture at various spatial-frequency scales may more accurately capture changes in tumor vascular heterogeneity as measured by DCE-MRI, and therefore provide better early prediction of NACT response.

Expression profile of CADM1 and CADM4 in triple negative breast cancer with primary systemic therapy.

Triple negative breast cancer (TNBC) is defined by a lack of ER, PgR, and HER2 expression, and to date there have been no significant advances in treatment by targeted therapies against those molecules. Therefore, primary systemic therapy (PST) followed by surgery is the standard therapy for patients with advanced TNBC. According to gene expression analysis, TNBC has a distinct profile when compared with non-TNBC, suggesting that a unique gene affects the treatment efficacy of PST. Cell adhesion molecule (CADM) genes encode an immunoglobulin superfamily molecule involved in cell-to-cell adhesion in a variety of human epithelial cells. While it has been reported that inactivation of CADM1 and CADM4 serves a pivotal role in the progression of breast cancer, a full analysis has not been completed for TNBC. Previous studies have reported that CADM1 and CADM4 expression is less likely to be decreased in TNBC than in non-TNBC. In the present study, CADM1 and CADM4 expression was evaluated in patients with TNBC who had received PST. The present study revealed that loss or weak expression of CADM1 was frequently observed in non-pathological complete response patients. Furthermore, while the majority of TNBC cases exhibited high CADM1 expression, a small number of cases exhibited low CADM1 expression and low therapeutic response of PST for TNBC. These results suggest that CADM1 has a pivotal role in anti-PST efficacy in patients with TNBC.

CCL20 triggered by chemotherapy hinders the therapeutic efficacy of breast cancer.

Chemotherapeutic resistance in triple-negative breast cancer (TNBC) has brought great challenges to the improvement of patient survival. The mechanisms of taxane chemoresistance in TNBC have not been well investigated. Our results illustrated C-C motif chemokine ligand 20 (CCL20) was significantly elevated during taxane-containing chemotherapy in breast cancer patients with nonpathologic complete response. Furthermore, CCL20 promoted the self-renewal and maintenance of breast cancer stem cells (BCSCs) or breast cancer stem-like cells through protein kinase Cζ (PKCζ) or p38 mitogen-activated protein kinase (MAPK)-mediated activation of p65 nuclear factor kappa B (NF-κB) pathway, significantly increasing the frequency and taxane resistance of BCSCs. Moreover, CCL20-promoted NF-κB activation increased ATP-binding cassette subfamily B member 1 (ABCB1)/multidrug resistance 1 (MDR1) expression, leading to the extracellular efflux of taxane. These results suggested that chemotherapy-induced CCL20 mediated chemoresistance via up-regulating ABCB1. In addition, NF-κB activation increased CCL20 expression, forming a positive feedback loop between NF-κB and CCL20 pathways, which provides sustained impetus for chemoresistance in breast cancer cells. Our results suggest that CCL20 can be a novel predictive marker for taxane response, and the blockade of CCL20 or its downstream pathway might reverse the taxane resistance in breast cancer patients.

High levels of serum CA15-3 and residual invasive tumor size are associated with poor prognosis for breast cancer patients with non-pathological complete response after neoadjuvant chemotherapy.

To identify surrogate markers for prognosis of breast cancer patients with non-pathological complete response (non-pCR) to neoadjuvant chemotherapy (NAC), our investigation focused on the serum levels of carcinoembryonic antigen (CEA) and carbohydrate antigen (CA15-3) as well as clinicopathological factors both before and after NAC. A total of 185 breast cancer patients treated with NAC were recruited. Serum carcinoembryonic antigen and CA15-3 were measured at baseline and at completion of NAC. Among the non-pCR cancers (n = 142), the disease-free survival (DFS) of patients with CA15-3-low at baseline (3-year DFS: 0.908, n = 73) was significantly better than of those with CA15-3-high (3-year DFS: 0.681, n = 69, P = .0134). Multivariable analysis demonstrated that baseline CA15-3 levels (hazard ratio: 3.31, 95% confidence interval: 1.28-10.23; P = .0122) and residual invasive size (hazard ratio: 4.47, 1.26-28.39; P = .0171) were significant independent factors for DFS. The combination of these factors proved to be an accurate predictor for DFS regardless of breast cancer subtypes. The combination of residual invasive size and serum CA15-3 levels at baseline seems to be a significant and independent surrogate marker of poor outcome for patients with non-pCR. These findings suggest that these markers may be useful for identifying patients with inferior prognosis and candidates for additional adjuvant treatments.

Digital gene expression profiling analysis and its application in the identification of genes associated with improved response to neoadjuvant chemotherapy in breast cancer

BackgroundThis study aimed to screen sensitive biomarkers for the efficacy evaluation of neoadjuvant chemotherapy in breast cancer.MethodsIn this study, Illumina digital gene expression sequencing technology was applied and differentially expressed genes (DEGs) between patients presenting pathological complete response (pCR) and non-pathological complete response (NpCR) were identified. Further, gene ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were then performed. The genes in significant enriched pathways were finally quantified by quantitative real-time PCR (qRT-PCR) to confirm that they were differentially expressed. Additionally, GSE23988 from Gene Expression Omnibus database was used as the validation dataset to confirm the DEGs.ResultsAfter removing the low-quality reads, 715 DEGs were finally detected. After mapping to KEGG pathways, 10 DEGs belonging to the ubiquitin proteasome pathway (HECTD3, PSMB10, UBD, UBE2C, and UBE2S) and cytokine–cytokine receptor interactions (CCL2, CCR1, CXCL10, CXCL11, and IL2RG) were selected for further analysis. These 10 genes were finally quantified by qRT-PCR to confirm that they were differentially expressed (the log2 fold changes of selected genes were − 5.34, 7.81, 6.88, 5.74, 3.11, 19.58, 8.73, 8.88, 7.42, and 34.61 for HECTD3, PSMB10, UBD, UBE2C, UBE2S, CCL2, CCR1, CXCL10, CXCL11, and IL2RG, respectively). Moreover, 53 common genes were confirmed by the validation dataset, including downregulated UBE2C and UBE2S.ConclusionOur results suggested that these 10 genes belonging to these two pathways might be useful as sensitive biomarkers for the efficacy evaluation of neoadjuvant chemotherapy in breast cancer.

Abstract P5-11-03: Tumor CXCL16/CXCR6 expression and soluble CXCL16 in HER2+ breast cancer (BC)

Abstract Background: CXCL16 is a pro-inflammatory chemokine associated with chemotaxis of CXCR6-expressing lymphocytes (T cells, NKT cells, NK cells) and the promotion of breast cancer cell proliferation, migration, and invasion in vitro. CXCL16 exists in a transmembrane or a cleaved, soluble form. There is limited clinically relevant data available on the CXCL16/CXCR6 signaling axis in HER2+ BC. This preliminary study examines tumor CXCL16 and CXCR6 mRNA expression and patient outcome in publicly available datasets and examines soluble CXCL16 in the plasma of 32 HER2+ BC patients enrolled in ICORG 10-05 (neo-adjuvant chemotherapy (docetaxel/carboplatin) +/- trastuzumab, lapatinib or trastuzumab/lapatinib). Methods: CXCL16 and CXCR6 mRNA expression was interrogated in publicly available datasets using BreastMark, a web-based tool for preliminary assessment of putative biomarkers in breast cancer.A median cut-off was used for all analyses. Pre-treatment and post-treatment (2 weeks pre-surgery) blood samples were collected from patients enrolled in ICORG 10-05. Plasma CXCL16 levels were determined by Luminex xMAP assay. Pre- and post-treatment levels of CXCL16 were compared directly or based on response (pathological complete response, CR, n=14 or non-pathological complete response, nCR, n=18). Stromal lymphocyte (SL) and tumor-infiltrating lymphocyte (TIL) levels were determined from Haematoxylin and Eosin-, AE1/AE3- and CD45FFPE- stained formalin-fixed paraffin embedded tissue. Pre-treatment lymphocyte levels were correlated with pre- and post-treatment levels of CXCL16 (Pearson's product-moment correlation). Results: In BC as a whole, analysis of publicly available datasets reveals tumor CXCL16 expression is not associated with outcome (n=1238, HR=0.9953, p=0.9516) but high tumor CXCR6 expression is (n=2652, HR=1.127, p=0.0476). Within a HER2+ cohort, inverse correlative analysis suggests high CXCR6/low CXCL16 tumor expression is significantly associated with a worse outcome (n=61, HR=2.731, p=0.01). For ICORG 10-05 patients, circulating CXCL16 plasma levels were significantly higher post-treatment (p&amp;lt;0.0001). The magnitude of this increase was significantly greater in CR than nCR patients (p&amp;lt;0.009). Post-treatment circulating CXCL16 levels negatively correlate with pre-treatment total (SL+TIL) lymphocyte counts (correlation coefficient -0.50 (CI -0.75- -0.13), p=0.01) in ICORG 10-05 patients. SL and total lymphocyte (SL+TIL) counts were higher in CR (n=13) vs. nCR (n=13) patients but the difference was not significant (SL, p=0.22; SL+TIL, p=0.29). Conclusions: Our preliminary results suggest tumor levels of CXCL16/CXCR6 are associated with patient outcome and circulating levels of soluble CXCL16 are altered by treatment and correlate with tumor immune infiltrate in HER2+ BC patients. Further examination of tumor CXCL16/CXCR6 expression and circulating CXCL16 as potential biomarkers of response is warranted in a larger cohort of HER2+ BC patients. Citation Format: Collins DM, Madden SF, Eustace AJ, Toomey S, Kay EW, Fay J, O'Donovan N, Gallagher WM, Hennessy B, Crown J. Tumor CXCL16/CXCR6 expression and soluble CXCL16 in HER2+ breast cancer (BC) [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 P5-11-03.

Change in HER2 status after neoadjuvant chemotherapy and the prognostic impact in patients with primary breast cancer.

We aimed to assess change in HER2 status after neoadjuvant chemotherapy (NAC) in patients with primary breast cancer and the prognostic impact of such changes. The study comprised 588 patients with a non-pathologic complete response who received anthracycline and/or taxane-based regimens in NAC setting. HER2 status was assessed before NAC and in residual invasive tumor of the surgical specimens. The associations between the change in HER2 status and clinicopathological factors were assessed. Before NAC, 489 (83%) of the 588 patients had HER2-negative tumors and 99 patients (17%) had HER2-positive tumors. Eleven (2.2%) of the HER2-negative tumors changed to HER2-positive, while 33 (33%) of the HER2-positive tumors changed to HER2-negative. ER and PR-positivity before NAC were associated with loss of HER2-positivity, whereas receiving trastuzumab was not. In terms of disease-free survival, there was no difference between patients with and those without change in HER2 status after NAC in either the patients with HER2-negative tumors (P = 0.26) or with HER2-positive tumors before NAC (P = 0.23). Our results showed that changes in HER2 status did not affect patients' prognosis. Further studies are needed to determine whether HER2-targeting agents can be omitted when loss of HER2-positivity is confirmed after NAC.

Texture analysis based on contrast-enhanced MRI can predict treatment response to neoadjuvant chemotherapy of breast cancer

Objective: To investigate whether texture analysis based on contrast-enhanced MRI can predict pathological complete response of locally advanced breast cancer undergoing neoadjuvant chemotherapy(NAC). Methods: Forty-seven patients with breast cancer undergone neoadjuvant chemotherapy from January 2015 to February 2016 were divided into pathological complete response (pCR) group or non-pathological complete response (non-pCR) group based on surgical pathology. Their parameters of texture analysis based on MRI before neoadjuvant chemotherapy and after 2 cycles of treatment were analyzed. Parameters(Energy, Entropy, Inertia, Correlation, Inverse Difference Moment)before and after 2 cycles of NAC between pCR and non-pCR groups were compared using Student t or Wilcoxon rank sum test. The diagnostic performance of different parameters was judged by the receiver-operating characteristic (ROC) curve analysis. Results: The post-NAC value was significantly different from that of pre-NAC (all P<0.05). Pre-treatment parameters (Energy, Entropy, Inertia, Correlation, Inverse Difference Moment) were 78.58×10(-5)(55.64×10(-5), 135.23×10(-5)), 10.06 ± 1.02, 7 993.91±2 428.10, (4.76±0.99) ×10(-5) and (18.10±4.13) ×10(-3) in pCR group, and 76.84×10(-5) (48.68×10(-5), 154.15×10(-5)), 10.28±1.26, 7 184.77 (4 938.03, 9 974.04), (5.21±2.01) ×10(-5) and (17.68±5.87) ×10(-3) in non-pCR group. No significant difference was found between both groups. (P>0.05 for all). At the end of the second cycle of NAC, parameters(Energy, Entropy, Inertia, Correlation, Inverse Difference Moment) were (542.11±361.04) ×10(-5,) 7.95±1.28, 16 765.08±97 06.56, (0.43±0.07) ×10(-5,) and (12.18±9.82) ×10(-3) in pCR group, and 133.00×10(-5) (79.80×10(-5,) 239.00×10(-5)), 9.29±1.46, 7 916.64(6 418.89, 10 934.40), (0.38±0.08) ×10(-5) and (14.80±5.06) ×10(-3) in non-pCR group. At the end of the second cycle of NAC, there was significant difference in the parameters (Energy, Entropy, Inertia, Correlation) and Δparameters (ΔEnergy, ΔEntropy, ΔInertia, ΔInverse Difference Moment) between both groups (P<0.05 for all). The area under curve (AUC) of post-treatment ΔEntropy was 0.81, which was the largest one among parameters. Sensitivity of ΔEntropy for predicting pCR was 75.0% and specificity was 85.7%, respectively. Conclusion: Texture analysis based on dynamic contrast-enhanced MRI can predict early treatment response in primary breast cancer.

Resistance to Taxanes in Triple-Negative Breast Cancer Associates with the Dynamics of a CD49f+ Tumor-Initiating Population.

SummaryTaxanes are a mainstay of treatment for breast cancer, but resistance often develops followed by metastatic disease and mortality. Aiming to reveal the mechanisms underlying taxane resistance, we used breast cancer patient-derived orthoxenografts (PDX). Mimicking clinical behavior, triple-negative breast tumors (TNBCs) from PDX models were more sensitive to docetaxel than luminal tumors, but they progressively acquired resistance upon continuous drug administration. Mechanistically, we found that a CD49f+ chemoresistant population with tumor-initiating ability is present in sensitive tumors and expands during the acquisition of drug resistance. In the absence of the drug, the resistant CD49f+ population shrinks and taxane sensitivity is restored. We describe a transcriptional signature of resistance, predictive of recurrent disease after chemotherapy in TNBC. Together, these findings identify a CD49f+ population enriched in tumor-initiating ability and chemoresistance properties and evidence a drug holiday effect on the acquired resistance to docetaxel in triple-negative breast cancer.

Early change of MR diffusion weighted imaging parameters of breast cancer after neoadjuvant chemotherapy and the relationship of the change with therapy response

Objective To investigate the early change of MR DWI parameters of breast cancer after neoadjuvant chemotherapy (NAC) and assess the relationship between the changes of the NAC response. Methods Patients with local advanced breast cancer diagnosed by biopsy were enrolled in the prospective study. They received DWI examination before NAC and then received DWI examination at 24 h after the first cycle of NAC. The pathological results of breast cancers were recorded after NAC finished. 14 patients (15 lesions) were enrolled in this study.The minimum ADC values of breast cancer before NAC and after the first cycle of NAC were recorded, and their change rates were calculated. Two weeks after NAC,all cases were divided into clinical complete response (cCR) and non-clinical complete response group according to clinical NAC response evaluation. Then lesions were divided into pathological complete response (pCR) and non-pathological complete response group according to pathological NAC response evaluation. Using the Paired samples t test,we analysed the change of ADC values 24 h after the first cycle of NAC. The ADC values and change rates were compared between cCR and non-cCR group using Mann-Whitney U test. The ADC values and change rates were also compared between pCR and non-pCR group using Mann-Whitney U test. Results The 15 cases were all invasive ductal carcinoma patients. According to clinical NAC response evaluation, 5 cases were cCR and 10 cases were non-cCR. According to pathological NAC response evaluation, 6 cases were pCR and 9 cases were non-pCR. Before NAC, the minimum ADC value of breast cancer was(0.78±0.15)×10-3 mm2/s. After the first cycle of NAC, the minimum ADC value of breast cancer was(0.91±0.16)×10-3 mm2/s. The minimum ADC value of breast cancer significantly increased 24 h after the first cycle of NAC (t=-2.954,P=0.010). But ADC value and its change rate were not significantly different between cCR and non-cCR groups, also between pCR and non-pCR groups (P>0.05). Conclusion Significant change in the minimum ADC value of breast cancer could be observed 24 h after the first cycle of NAC, but the change do not relate to clinical and pathological NAC response in this study. Key words: Breast neoplasms; Magnetic resonance imaging; Treatment outcome

RNA Sequencing Uncovers Molecular Mechanisms Underlying Pathological Complete Response to Chemotherapy in Patients with Operable Breast Cancer.

BackgroundThis study aimed to identify key genes contributing to pathological complete response (pCR) to chemotherapy by mRNA sequencing (RNA-seq).Material/MethodsRNA was extracted from the frozen biopsy tissue of patients with pathological complete response and patients with non-pathological complete response. Sequencing was performed on the HiSeq2000 platform. Differentially expressed genes (DEGs) were identified between the pCR group and non-pCR (NpCR) group. Pathway enrichment analysis of DEGs was performed. A protein-protein interaction network was constructed, then module analysis was performed to identify a subnetwork. Finally, transcription factors were predicted.ResultsA total of 673 DEGs were identified, including 419 upregulated ones and 254 downregulated ones. The PPI network constructed consisted of 276 proteins forming 471 PPI pairs, and a subnetwork containing 18 protein nodes was obtained. Pathway enrichment analysis revealed that PLCB4 and ADCY6 were enriched in pathways renin secretion, gastric acid secretion, gap junction, inflammatory mediator regulation of TRP channels, retrograde endocannabinoid signaling, melanogenesis, cGMP-PKG signaling pathway, calcium signaling pathway, chemokine signaling pathway, cAMP signaling pathway, and rap1 signaling pathway. CNR1 was enriched in the neuroactive ligand-receptor interaction pathway, retrograde endocannabinoid signaling pathway, and rap1 signaling pathway. The transcription factor-gene network consists of 15 transcription factors and 16 targeted genes, of which 5 were downregulated and 10 were upregulated.ConclusionsWe found key genes that may contribute to pCR to chemotherapy, such as PLCB4, ADCY6, and CNR1, as well as some transcription factors.

Role of the Intravoxel Incoherent Motion Diffusion Weighted Imaging in the Pre-treatment Prediction and Early Response Monitoring to Neoadjuvant Chemotherapy in Locally Advanced Breast Cancer.

The aim of this study was to explore whether intravoxel incoherent motion (IVIM) diffusion-weighted imaging (DWI) can probe pre-treatment differences or monitor early response in patients with locally advanced breast cancer receiving neoadjuvant chemotherapy (NAC).Thirty-six patients with locally advanced breast cancer were imaged using multiple-b DWI with 12 b values ranging from 0 to 1000 s/mm2 at the baseline, and 28 patients were repeatedly scanned after the second cycle of NAC. Subjects were divided into pathologic complete response (pCR) and nonpathologic complete response (non-pCR) groups according to the surgical pathologic specimen. Parameters (D, D∗, f, maximum diameter [MD] and volume [V]) before and after 2 cycles of NAC and their corresponding change (Δparameter) between pCR and non-pCR groups were compared using the Student t test or nonparametric test. The diagnostic performance of different parameters was judged by the receiver-operating characteristic curve analysis.Before NAC, the f value of pCR group was significantly higher than that of non-pCR (32.40% vs 24.40%, P = 0.048). At the end of the second cycle of NAC, the D value was significantly higher and the f value was significantly lower in pCR than that in non-pCR (P = 0.001; P = 0.015, respectively), whereas the D∗ value and V of the pCR group was slightly lower than that of the non-pCR group (P = 0.507; P = 0.676, respectively). ΔD was higher in pCR (−0.45 × 10–3 mm2/s) than that in non-pCR (−0.07 × 10−3 mm2/s) after 2 cycles of NAC (P < 0.001). Δf value in the pCR group was significantly higher than that in the non-pCR group (17.30% vs 5.30%, P = 0.001). There was no significant difference in ΔD∗ between the pCR and non-pCR group (P = 0.456). The prediction performance of ΔD value was the highest (AUC [area under the curve] = 0.924, 95% CI [95% confidence interval] = 0.759–0.990). When the optimal cut-off was set at −0.163 × 10−3 mm2/s, the values for sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were up to 100% (95% CI = 66.4–100), 73.7% (95% CI = 48.8–90.9), 64.3% (95% CI = 35.6–86.0), and 100% (95% CI = 73.2–99.3), respectively.IVIM-derived parameters, especially the D and f value, showed potential value in the pre-treatment prediction and early response monitoring to NAC in locally advanced breast cancer. ΔD value had the best prediction performance for pathologic response after NAC.

Prognostic significance of tumor-infiltrating CD8+ and FOXP3+ lymphocytes in residual tumors and alterations in these parameters after neoadjuvant chemotherapy in triple-negative breast cancer: a retrospective multicenter study

IntroductionThe status of tumor-infiltrating lymphocytes (TILs) has been recently proposed to predict clinical outcome of patients with breast cancer. We therefore studied the prognostic significance of CD8+ TILs and FOXP3+ TILs in residual tumors after neoadjuvant chemotherapy (NAC) and the alterations in these parameters before and after NAC in patients with triple-negative breast cancer (TNBC).MethodsOne hundred thirty-one TNBC patients who received NAC at three institutions were examined. CD8+ TIL and FOXP3+ TIL in residual tumors and biopsy specimens were evaluated by double-staining immunohistochemistry. The CD8+ TIL and FOXP3+ TIL status of the residual tumors was assessed, and the rates of their changes before and after NAC were calculated.ResultsTNBC patients with high CD8+ TIL levels or a high CD8/FOXP3 ratio in residual tumors had significantly better recurrence-free survival (RFS) and breast cancer-specific survival (BCSS) than patients with low values of these parameters. In multivariate analyses, CD8+ TIL exhibited strong prognostic significance for RFS, with a hazard ratio (HR) of 3.09 (95 % confidence interval (CI) 1.537–6.614, P=0.0013). The CD8/FOXP3 ratio was also significantly correlated with RFS (HR=2.07, 95 % CI 1.029–4.436, P=0.0412). TNBC with larger residual tumor size and positive lymph node status, which are known prognostic factors, was independently associated with worse RFS (P=0.0064 and P=0.0015, respectively). High CD8+ TIL levels were a markedly powerful indicator of improved BCSS, with an HR of 3.59 (95 % CI 1.499–9.581, P=0.0036). Nodal status was also associated with BCSS (P=0.0024). TNBC with a high rate of CD8+ TIL changes was associated with significantly better RFS compared with the low group (P=0.011). Higher rates of changes in the CD8/FOXP3 ratio were significantly correlated with both better RFS and BCSS compared with lower rates (P=0.011 and P=0.023, respectively).ConclusionsThis is the first study to demonstrate that high CD8+ TIL and a high CD8/FOXP3 ratio in residual tumors and increment of these parameters following NAC and accurately predict improved prognosis in TNBC patients with non-pathological complete response following NAC. These parameters could serve as a surrogate one for adjuvant treatment in patients with residual disease in the neoadjuvant setting.Electronic supplementary materialThe online version of this article (doi:10.1186/s13058-015-0632-x) contains supplementary material, which is available to authorized users.

Impact of biomarker changes during neoadjuvant chemotherapy for clinical response in patients with residual breast cancers.

Residual cancer burden or Ki67 expression levels in residual tumors reportedly provided significant prognostic information for a non-pathological complete response subset after neoadjuvant chemotherapy (NAC). However, the significance of Ki67 reduction for clinical response during chemotherapy in each subtype or menopausal status is yet to be determined. A total of 183 breast cancers surgically removed after chemotherapy were recruited for this study. Expression levels of estrogen receptor (ER), progesterone receptor (PgR), and Ki67 were determined immunohistochemically for semiquantitative measurement and these biomarkers were compared in pre- and post-NAC samples from pathological non-responders (n = 125). Responses to chemotherapy were evaluated both clinically and pathologically. Ki67 expression levels after NAC (median 5 %, range 0-70 %) were significantly reduced compared with before NAC (25, 1-80 %, P < 0.0001), but only in patients who attained clinical response. This significant suppression of Ki67 in clinical responders was consistently observed in breast cancers from the ER-positive subset, but not the ER-negative subset in the total test set (n = 120). These observations were also made in the validation set (n = 63). Among premenopausal, but not postmenopausal patients, a significant decrease in PgR expression levels was detected in breast cancers of patients who attained clinical response (pre-NAC 50, 0-100 %, post-NAC 5, 0-20 %; P = 0.0003). The impact of Ki67 suppression on clinical response seems to be restricted to ER-positive breast cancers. Since PgR expression levels of premenopausal ER-positive cancers were significantly reduced in clinical responders, inhibition of estrogen signaling due to chemotherapy-induced amenorrhea may be involved in this association.

Axillary pathologic response after neoadjuvant chemotherapy in locally advanced breast cancer with axillary involvement

AimTo compare axillary involvement (N+) at initial staging in locally advanced breast cancer (LABC) with axillary lymphadenectomy histologic results after neoadjuvant chemotherapy treatment (NeoChemo). Material and methodsRetrospective study between November 2011 and September 2013 of LABC cases treated with neoadjuvant chemotherapy based on docetaxel (associated with trastuzumab in HER2-positive cases and carboplatin/adriamycin in HER2-negative cases). Those clinically or radiologically suspected cases of axillary involvement were histologically confirmed. When there was no suspicion of axillary involvement, sentinel lymph node radioguided biopsy (SLNRB) was performed using intradermal injection of 99mTc-nanocolloid albumin prior to neoadjuvant treatment. Axillary lymphadenectomy after NeoChemo was undertaken in all cases with positive axilla. Final pathologic response was classified as complete (pCR) when there was no evidence of tumoral disease and as non-pathologic complete response (no pCR) in the opposite case. ResultsA total of 346 patients treated with docetaxel were reviewed, identifying 105 LABC. Axillary involvement at initial staging was detected in 70 (67%) before starting NeoChemo. From these 70, 73% (n=51) were N+ (fine needle biopsy and/or biopsy) and the remaining 19 (27%) were occult N+ detected by SLNRB. Axillary lymphadenectomy detected pCR in 56% (39/70), increasing up to 84% pCR when initial N+ status was reached using SNLB. On the other hand, when N+ was detected using fine needle biopsy/lymph biopsy, pCR was only 45%. ConclusionMore than 50% of women affected by locally advanced breast cancer with tumoral axillary involvement at initial diagnosis present free metastatic axilla after therapeutic neoadjuvant chemotherapy effect. This increases up to almost 90% in case of occult metastatic axilla detected with sentinel node biopsy prior to starting neoadjuvant chemotherapy.

Abstract P6-01-03: Change of HER2 status following neoadjuvant systemic therapy in primary breast cancer patients

Abstract Background: The incidence of change in HER2 status in primary breast cancer after neoadjuvant chemotherapy (NAC) and whether the prognosis is affected by the change in HER2 status is not well known. Patients and Methods: Five hundred and eighty-eight patients who were treated with anthracycline- and/or taxane-based NAC and had non-pathologic complete response between 2001 and 2008 in our hospital were enrolled. Human epidermal growth factor receptor-2 (HER2) status was assessed in specimen by needle biopsy before NAC and on the residual tumor of surgical specimen. We determine the impact of change in HER2 status on recurrence-free survival (RFS). Twenty-eight patients had received trastuzumab with NAC, and 57 patients had received trastuzumab as adjuvant therapy. HER2-positive was defined as 3+ by immunohistochemistry and/or amplification by fluorescent in situ hybridization. Association between change in HER2 status after NAC and clinicopathologic factors, including age, clinical T stage, estrogen receptor (ER), progesterone receptor (PR), Nuclear grade (NG), lymphovascular invasion (LVI) and Trastuzumab usage (NAC and Adjuvant setting) were determined. Result: A median follow-up period was 57 months (range, 3 to 131 months). Four hundred eighty-nine of the 588 patients (83.1%) had HER2-negative tumors and 99 patients (16.8%) had HER2-positive tumor before NAC. In 11 of the 489 patients (2.2%) HER2-negative changed to HER2-positive.In 33 of the 99 patients (30%) HER2-positive changed to HER2-negative. In clinicopathologic factors, ER and PR positive before NAC were associated with incidence of change in HER2 status after NAC. Receiving trastuzumab was not correlated with incidence of change in HER2 status. In terms of RFS, there was no difference between patients with and without change in HER2 status in both of the 489 patients with HER2-negative tumors and 99 patients with HER2-positive tumors before NAC (p=0.26, p=0.23, respectively). Conclusion: We herein reported the incidence of change in HER2 status after NAC with the largest sample size. However, change in HER2 status did not seems to affect prognosis. Further prospective study is needed to confirm the prognostic impact of change in HER2 status. Citation Format: Atsushi Yoshida, Naoki Hayashi, Hiroshi Yagata, Koyu Suzuki, Seigo Nakamura, Hideko Yamauchi. Change of HER2 status following neoadjuvant systemic therapy in primary breast cancer patients [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 P6-01-03.

Prognostic significance of the ratio of absolute neutrophil to lymphocyte counts for breast cancer patients with ER/PR-positivity and HER2-negativity in neoadjuvant setting.

The aim of this study was to determine the predictive or prognostic impact of absolute neutrophil count/absolute lymphocyte count ratio (NLR) in breast cancer patients with estrogen receptor/progesterone receptor (ER/PR)-positive and human epidermal growth factor receptor 2 (HER2)-negative subtype who have received neoadjuvant chemotherapy (NAC). We performed retrospective analysis of 157 patients with primary breast cancer with ER/PR-positive and HER2-negative subtype who were treated with NAC, followed by definitive surgical resection. The median follow-up after surgery was 21 months (range, 1-108 months). On univariate analysis, high NLR (>2.25) correlated with poorer recurrence-free survival (RFS) and overall survival (OS) (P = 0.001 and P < 0.001). Subgroup analysis of non-pathologic complete response (pCR) subgroup showed that high NLR was significant for RFS and OS (P = 0.001 and P < 0.001). Particularly, high NLR patients had inferior clinical outcomes in the high clinical stage. Uni- and multivariate Cox analysis showed NLR to be an only predictor of RFS and OS. The NLR is an independent prognostic factor for RFS and OS in breast cancer patients with ER/PR-positive and HER2-negative subtype receiving NAC. The NLR provides additional prognostic information to choose suitable patients who might profit from further therapy.