1014 Background: Predicting recurrence in operable breast cancer (BC) despite optimal chemotherap... more 1014 Background: Predicting recurrence in operable breast cancer (BC) despite optimal chemotherapy would be relevant to new drug development and tailored treatments. Methods: A large series (n=3,154) of public Affymetrix gene-expression profiles (GEP) was used to define prognostic/predictive metagenes in different BC subtypes. In ER+/HER2- a proliferation and an ER-related metagene were combined to predict low, intermediate and high risk of recurrence. In TN and in HER2+ a T cell metagene was used to predict low, intermediate and high risk (higher expression associated with lower risk). The metagenes were validated in patients enrolled in the phase III ECTO trial (Gianni L. JCO 2009) and treated with the same taxane-anthracycline-CMF regimen as neoadjuvant or adjuvant therapy before endocrine therapy if indicated. The outcome was distant event free survival (DEFS). Results: 283 good quality GEPs were obtained (neoadjuvant n=121; adjuvant n=162) from 464 retrospectively collected samples. Median follow-up was 8.9 years. In ER+/HER2- tumors the 10-yrs DEFS was 92.3, 81.2 and 66.6% in low, intermediate and high risk groups, respectively [high vs low HR 4.38 (1.01-19.1) p=.048] according to proliferation and ER-related metagenes. In HER2+ and TN subgroup the 10-yrs DEFS was 97.2, 75.6 and 78.8% in low, intermediate and high risk groups, respectively [high vs low HR 8.73 (1.09-69.8) p=.041]. In TN tumors, the pCR rate was 20% in the high and 61.5% in the low risk group. By combining the predicted risk group in each molecular subtype the 10-yrs DEFS was 95.3, 79.2 and 71.5% in low (24.2%), intermediate (42.7%) and high (33.1%) risk group, respectively [logrank p=0.003; high vs low HR 6.22 (1.87-20.6) p=.002]. ER, PGR, Ki67 and lymphocyte infiltration (LI) by IHC underperformed compared to genomic predictors. Conclusions: BC patients at higher risk of relapse despite optimal standard treatment can be identified who should be spared ineffective and toxic therapy and considered for investigational new strategies. In TN and HER2+, high T cell metagene and to a lesser extent LI are prognostic/predictive and associated with an extremely low risk of DEFS after chemotherapy.
513 Background: In the mid 1990’s data indicated that paclitaxel had marked antitumor activity in... more 513 Background: In the mid 1990’s data indicated that paclitaxel had marked antitumor activity in metastatic breast cancer, justifying the attempt at investigating whether the taxane could improve the therapeutic benefit of established regimens in early breast cancer Methods: From 1996 to 2002 a total of 1355 women with operable breast cancer (T>2cm) were randomized to adjuvant A (75 mg/m2 q21d x 4) followed by i.v. CMF (day 1&8 q28d x 4), or adjuvant A (60 mg/m2) and T (200 mg/m2 over 3 hrs q21d x 4) followed by CMF (AT→CMF), or AT→CMF as primary systemic therapy (PST). Results: Main patient characteristics (T-size, ER/PgR status, grade and age) were evenly distributed among arms. After median follow-up of 43 months, freedom from progression (FFP) was significantly better for women receiving adjuvant AT→CMF than A→CMF (HR 0.65, range 0.48–0.90, P=0.01). In a multivariate analysis, treatment inclusive of paclitaxel stood out as significantly associated with FFP (HR 0.66, P=0.012), together with clinical d...
537 Background: At the time the ECTO was designed in 1996, taxanes were only indicated for patien... more 537 Background: At the time the ECTO was designed in 1996, taxanes were only indicated for patients with metastatic breast cancer. However, paclitaxel and docetaxel were still to be tested in the adjuvant setting. In addition there was relatively scarce information on the comparative efficacy of neoadjuvant and adjuvant regimens. The ECTO trial was designed to evaluate the addition of paclitaxel to an anthracycline-based adjuvant regimen and to compare this combination with the same regimen given as primary systemic (neoadjuvant) therapy. Methods: A total of 1,355 women with operable breast cancer were randomized to one of three treatments: 1) surgery followed by adjuvant single agent doxorubicin (A) followed by CMF (arm A); 2) surgery followed by adjuvant paclitaxel plus doxorubicin (AT) followed by CMF (arm B); 3) AT followed by CMF followed by surgery (arm C). The two co-primary objectives were to assess the effects on freedom from progression (FFP) of: 1) the addition of paclitaxel to post-operative chemotherapy (arm B versus arm A); and 2) primary versus adjuvant chemotherapy (arm B versus arm C). Results: At 10 years, in the adjuvant setting FFP remained statistically significant in favor of AT followed by CMF (arm B, HR 0.77, P=0.045). Distant FFP was similarly improved but overall survival was not (HR 0.82, P=0.24). There was no significant difference in FFP when chemotherapy was given after surgery compared with the same regimen given before surgery (arm B vs arm C, HR 0.79, P=0.07). In the primary chemotherapy arm, patients who achieved a pathological complete remission (pCR) had improved distant FFP (P < 0.001) compared to patients who did not achieve pCR. When given as primary systemic therapy, the paclitaxel-containing regimen allowed breast-sparing surgery in a significant percentage of patients, which did not translate in an increased risk of ipsilateral breast recurrence compared to the risk observed in patients in the adjuvant arms. Conclusions: Incorporating paclitaxel into anthracycline-based adjuvant therapy resulted in a significantly improved FFP and DFFP.
Trastuzumab treatment has significantly improved the prognosis of HER2-positive breast cancer pat... more Trastuzumab treatment has significantly improved the prognosis of HER2-positive breast cancer patients. Despite this, resistance to therapy still remains the main clinical challenge. In order to evaluate the implication of microRNAs in the trastuzumab response, we performed a microRNA array in parental and acquired trastuzumab-resistant HER2-positive breast cancer cell lines. Our results identified miR-146a-5p as the main dysregulated microRNA. Interestingly, high miR-146a-5p expression in primary tumor tissue significantly correlated with shorter disease-free survival in HER2-positive breast cancer patients. The gain- and loss-of-function of miR-146a-5p modulated the response to trastuzumab. Furthermore, the overexpression of miR-146a-5p increased migration and angiogenesis, and promoted cell cycle progression by reducing CDKN1A expression. Exosomes from trastuzumab-resistant cells showed a high level of miR-146a-5p expression compared with the parental cells. In addition, the co-c...
Anti-HER2 therapies have markedly improved prognosis of HER2-positive breast cancer. However, dif... more Anti-HER2 therapies have markedly improved prognosis of HER2-positive breast cancer. However, different mechanisms play a role in treatment resistance. Here, we identified AXL overexpression as an essential mechanism of trastuzumab resistance. AXL orchestrates epithelial-to-mesenchymal transition and heterodimerizes with HER2, leading to activation of PI3K/AKT and MAPK pathways in a ligand-independent manner. Genetic depletion and pharmacological inhibition of AXL restored trastuzumab response in vitro and in vivo. AXL inhibitor plus trastuzumab achieved complete regression in trastuzumab-resistant patient-derived xenograft models. Moreover, AXL expression in HER2-positive primary tumors was able to predict prognosis. Data from the PAMELA trial showed a change in AXL expression during neoadjuvant dual HER2 blockade, supporting its role in resistance. Therefore, our study highlights the importance of targeting AXL in combination with anti-HER2 drugs across HER2-amplified breast cance...
Novel antibody-drug conjugates against HER2 are showing high activity in HER2-negative breast can... more Novel antibody-drug conjugates against HER2 are showing high activity in HER2-negative breast cancer (BC) with low HER2 expression (i.e., 1+ or 2+ and lack of ERBB2 amplification). However, the clinical and molecular features of HER2-low BC are yet to be elucidated. Here, we collected retrospective clinicopathological and PAM50 data from 3,689 patients with HER2-negative disease and made the following observations. First, the proportion of HER2-low was higher in HR-positive disease (65.4%) than triple-negative BC (TNBC, 36.6%). Second, within HR-positive disease, ERBB2 and luminal-related genes were more expressed in HER2-low than HER2 0. In contrast, no gene was found differentially expressed in TNBC according to HER2 expression. Third, within HER2-low, ERBB2 levels were higher in HR-positive disease than TNBC. Fourth, HER2-low was not associated with overall survival in HR-positive disease and TNBC. Finally, the reproducibility of HER2-low among pathologists was suboptimal. This s...
532 Background: NOAH (NeOAdjuvant Herceptin) is a Phase III trial of neoadjuvant trastuzumab; H) ... more 532 Background: NOAH (NeOAdjuvant Herceptin) is a Phase III trial of neoadjuvant trastuzumab; H) in combination with chemotherapy in patients (pts) with HER2-positive locally advanced breast cancer (LABC). Methods: 228 pts with centrally confirmed HER2-positive (IHC 3+ or FISH+) LABC received 3 cycles of doxorubicin-paclitaxel (AT: A 60 mg/m2, T 150 mg/m2 q3w), 4 cycles of T (175 mg/m2 q3w) and 3 cycles of cyclophosphamide/methotrexate/5-fluorouracil (CMF: C 600 mg/m2, M 40 mg/m2, F 600 mg/m2 q4w) on days 1 and 8, with (n=115) or without (n=113) concomitant H (8 mg/kg loading dose then 6 mg/kg q3w for 1 year) before surgery. Pts with HER2- negative disease (IHC 0/1+; n=99) were treated in parallel with AT/T/CMF. The primary end point was event-free survival (EFS); secondary end points included overall response rate (ORR), pathological complete response (pCR) rate and safety. Results: Baseline characteristics were well balanced for randomised pts. Median tumour size was 5.5 cm (range...
Natural killer (NK) cells can orchestrate effective antitumor immunity. The presence of tumor-inf... more Natural killer (NK) cells can orchestrate effective antitumor immunity. The presence of tumor-infiltrating NK cells in diagnostic biopsies predicts pathologic complete response (pCR) to HER2-specific therapeutic antibodies in patients with primary breast cancer. Here, we analyzed whether diversity in circulating NK cells might influence tumor infiltration and HER2-specific therapeutic antibody efficacy. We found that numbers of circulating CD57+ NK cells inversely correlated with pCR to HER2-specific antibody treatment in patients with primary breast cancer independently of age, traditional clinicopathologic factors, and CD16A 158F/V genotype. This association was uncoupled from the expression of other NK-cell receptors, the presence of adaptive NK cells, or changes in major T-cell subsets, reminiscent of cytomegalovirus-induced immunomodulation. NK-cell activation against trastuzumab-coated HER2+ breast cancer cells was comparable in patients with high and low proportions of CD57+ ...
Purpose: This study aimed to identify biomarkers of resistance to endocrine therapy in estrogen r... more Purpose: This study aimed to identify biomarkers of resistance to endocrine therapy in estrogen receptor–positive (ER+) breast cancers treated with prolonged neoadjuvant letrozole.Experimental Design: We performed targeted DNA and RNA sequencing in 68 ER+ breast cancers from patients treated with preoperative letrozole (median, 7 months).Results: Twenty-four tumors (35%) exhibited a PEPI score ≥4 and/or recurred after a median of 58 months and were considered endocrine resistant. Integration of the 47 most upregulated genes (log FC > 1, FDR < 0.03) in letrozole-resistant tumors with transcription-binding data showed significant overlap with 20 E2F4-regulated genes (P = 2.56E−15). In patients treated with the CDK4/6 inhibitor palbociclib before surgery, treatment significantly decreased expression of 24 of the 47 most upregulated genes in letrozole-resistant tumors, including 18 of the 20 E2F4 target genes. In long-term estrogen-deprived ER+ breast cancer cells, palbociclib als...
BackgroundNeoadjuvant treatment is increasingly one of the preferred therapeutic options for earl... more BackgroundNeoadjuvant treatment is increasingly one of the preferred therapeutic options for early breast cancer and may have some unique outcomes, such as identifying predictive and prognostic factors of response or increasing the knowledge of individual tumor biology.DesignA panel of experts from different specialties reviewed published clinical studies on the neoadjuvant management of breast cancer. Recommendations were made that emphasized the clinical multidisciplinary management and the investigational leverage in early breast cancer.ResultsNeoadjuvant therapy has equivalent efficacy to adjuvant therapy, and it has some additional benefits that include increasing breast conservation, assessing tumor response, establishing prognosis based on the pathological response, and providing a “second opportunity” for nonresponding patients. Achieving pathological complete remission because of neoadjuvant therapy has been correlated with long-term clinical benefit, particularly in HER2-p...
Clinical cancer research : an official journal of the American Association for Cancer Research, Jan 6, 2018
We investigated the value of tumor-infiltrating NK cells (TI-NK) and HLA class I tumor expression... more We investigated the value of tumor-infiltrating NK cells (TI-NK) and HLA class I tumor expression as biomarkers of response to neoadjuvant anti-HER2 antibody-based treatment in breast cancer. TI-NK cells and HLA-I were determined by immunohistochemistry in pre-treatment tumor biopsies from two cohorts of HER2-positive breast cancer patients [discovery cohort (n=42) and validation cohort (n=71)]. TIL were scored according to international guidelines. Biomarker association with pathological complete response (pCR) and disease-free survival (DFS) was adjusted for prognostic factors. Gene set variation analysis was used for determining immune-cell populations concomitant to NK cell enrichment in HER2-positive tumors from the TCGA (n=190). TI-NK cells were significantly associated with pCR in the discovery cohort as well as in the validation cohort (p<0.0001), independently of clinicopathological factors. A ≥3 TI-NK cells/50xHPF cut off predicted pCR in the discovery and validation co...
554 Background: Gene expression profiling can be used to predict the likelihood of achieving a pa... more 554 Background: Gene expression profiling can be used to predict the likelihood of achieving a pathological complete response (pCR) to neoadjuvant chemotherapy (CT) in patients with primary breast cancer. We report a retrospective exploratory analysis to evaluate the utility of the PAM50 subtype assay in predicting pCR in women enrolled in the NeOAdjuvant Herceptin (NOAH) trial, which recruited 334 patients. METHODS Formalin-fixed paraffin-embedded core biopsies were collected from 156 patients randomized to receive doxorubicin/paclitaxel (AT) followed by CMF (Group 1; HER2-positive/negative), or the same regimen with trastuzumab (H) (AT+H→CMF+H; Group 2; HER2-positive). Total RNA was amplified, labeled, and hybridized onto Affymetrix 2.0 Plus microarrays. The PAM50 microarray-based assay was applied to 43 genes. The PAM50 Risk Of Relapse score based on subtype and Proliferation (RORP) was evaluated as a continuous variable and as low/medium/high-risk groups using previously reported cutoffs. pCR was defined by the absence of residual invasive tumor in the breast and axillary lymph nodes. Univariate logistic regression was used to evaluate the association of each signature with pCR. RESULTS Overall pCR rates were 21.5% (20/93) and 44.4% (28/63) in Groups 1 and 2, respectively (p<0.005). In Group 1, non-LumA tumors showed higher pCR rates than LumA tumors (27% vs. 0%, p<0.005). Low, medium and high RORP groups showed 0%, 19% and 37% pCR rates (p=0.02). In Group 2, all subtypes were identified and HER2-enriched tumors showed a non-significant increase in pCR rates compared to non-HER2-enriched tumors (53% vs. 34%). Low, medium and high RORP groups showed 17%, 36% and 75% pCR rates (p=0.01). In Group 2 the odds ratio for achieving a pCR in the high-risk RORP group was 15.0 compared to the low-risk group (p=0.029). CONCLUSIONS HER2-positive tumors predicted to have a high risk of relapse by the PAM50 assay benefited substantially from H-based CT. Similar findings are observed in HER2-positive/negative tumors treated with CT only. The role of PAM50 and/or other genomics assays in predicting pCR and survival during treatment with anti-HER2 agents warrants further investigation.
529 Background: HER2-positive/ER+ and HER2-positive/ER- breast cancers have different patterns of... more 529 Background: HER2-positive/ER+ and HER2-positive/ER- breast cancers have different patterns of gene expression and intracellular pathway activation. We assessed the role of selected biomarkers in predicting the likelihood of achieving a pathological complete response (pCR) in the NeOAdjuvant Herceptin (NOAH) trial, which recruited 334 patients. METHODS Formalin-fixed paraffin-embedded core biopsies were collected from 114 women randomized to receive trastuzumab (H) with doxorubicin/paclitaxel followed by CMF (Arm CTH) or chemotherapy alone (Arm CT). Expression of the following genes and metagenes was profiled on Affymetrix 2.0 Plus: MYC, PIK3CA, PIK3R1, PTEN, amphiregulin (AREG), epiregulin, 8q24 and 8q22 amplicons, plasma cell (PC) and IGF metagenes. ER status was based on ESR1 gene expression. pCR was defined as the absence of residual invasive tumor in the breast and axillary lymph nodes. Biomarkers were assessed as continuous variables by univariate logistic regression analysis. RESULTS 80 samples (70%) were ER- and 34 (30%) were ER+. In the CT arm AREG (p=0.03) and 8q22 (p=0.06) were associated with lower pCR rates among ER- samples, while PTEN (p=0.01) was associated with higher pCR rates. Among ER- samples in the CTH arm, AREG (p=0.04) and IGF (p=0.007) were associated with resistance, while PC showed a trend for higher pCR rate (p=0.079). There were significant interactions between the treatment arms and 8q22 (p=0.03) and PC (p=0.04). In the high tertile of 8q22 the pCR rates were 0% (0/12 Arm CT) and 67% (10/15 Arm CTH) while for PC the pCR rates were 13% (2/15 Arm CT) and 83% (10/12 Arm CTH). There were no statistically significant associations for ER+ tumors, but a significant interaction for the likelihood of pCR and ER status was observed for PTEN (p=0.006) and PC (p=0.05) in the CT Arm and for AREG (p=0.03) in the CTH Arm. CONCLUSIONS High expression of the PC metagene and the 8q22 amplicon, and low expression of the IGF metagene, were associated with the increased pCR rate reported with the addition of H to CT in HER2-positive/ER- tumors. The concept that HER2-positive/ER+ and HER2-positive/ER- tumors are driven by different biologic pathways warrants further investigation.
Clinical cancer research : an official journal of the American Association for Cancer Research, 2002
The full-length receptor p185HER-2 undergoes a metalloprotease-dependent cleavage producing a mem... more The full-length receptor p185HER-2 undergoes a metalloprotease-dependent cleavage producing a membrane-associated fragment (p95HER-2) in cultured breast cancer cells. P95HER-2 has potentially enhanced signaling activity, but its expression and role in human breast cancer is poorly characterized. The purpose of this project was to characterize the expression of p95HER-2 in primary breast cancers and nodal metastasis, and to study association with clinicopathological factors. P95HER-2 and p185HER-2 were examined in 337 primary breast tumors and 81 metastatic lymph nodes by Western blot analysis, and tested for associations with other clinicopathological factors. P95HER-2 was present in 20.9% of primary tumors from node-negative patients, in 29.1% from patients with one to three metastatic nodes, and in 36.7% from patients with four or more metastatic nodes (P = 0.027). Whereas p185HER-2 overexpression was unrelated to nodal disease (P = 0.63), the odds of lymph node metastasis were en...
1014 Background: Predicting recurrence in operable breast cancer (BC) despite optimal chemotherap... more 1014 Background: Predicting recurrence in operable breast cancer (BC) despite optimal chemotherapy would be relevant to new drug development and tailored treatments. Methods: A large series (n=3,154) of public Affymetrix gene-expression profiles (GEP) was used to define prognostic/predictive metagenes in different BC subtypes. In ER+/HER2- a proliferation and an ER-related metagene were combined to predict low, intermediate and high risk of recurrence. In TN and in HER2+ a T cell metagene was used to predict low, intermediate and high risk (higher expression associated with lower risk). The metagenes were validated in patients enrolled in the phase III ECTO trial (Gianni L. JCO 2009) and treated with the same taxane-anthracycline-CMF regimen as neoadjuvant or adjuvant therapy before endocrine therapy if indicated. The outcome was distant event free survival (DEFS). Results: 283 good quality GEPs were obtained (neoadjuvant n=121; adjuvant n=162) from 464 retrospectively collected samples. Median follow-up was 8.9 years. In ER+/HER2- tumors the 10-yrs DEFS was 92.3, 81.2 and 66.6% in low, intermediate and high risk groups, respectively [high vs low HR 4.38 (1.01-19.1) p=.048] according to proliferation and ER-related metagenes. In HER2+ and TN subgroup the 10-yrs DEFS was 97.2, 75.6 and 78.8% in low, intermediate and high risk groups, respectively [high vs low HR 8.73 (1.09-69.8) p=.041]. In TN tumors, the pCR rate was 20% in the high and 61.5% in the low risk group. By combining the predicted risk group in each molecular subtype the 10-yrs DEFS was 95.3, 79.2 and 71.5% in low (24.2%), intermediate (42.7%) and high (33.1%) risk group, respectively [logrank p=0.003; high vs low HR 6.22 (1.87-20.6) p=.002]. ER, PGR, Ki67 and lymphocyte infiltration (LI) by IHC underperformed compared to genomic predictors. Conclusions: BC patients at higher risk of relapse despite optimal standard treatment can be identified who should be spared ineffective and toxic therapy and considered for investigational new strategies. In TN and HER2+, high T cell metagene and to a lesser extent LI are prognostic/predictive and associated with an extremely low risk of DEFS after chemotherapy.
513 Background: In the mid 1990’s data indicated that paclitaxel had marked antitumor activity in... more 513 Background: In the mid 1990’s data indicated that paclitaxel had marked antitumor activity in metastatic breast cancer, justifying the attempt at investigating whether the taxane could improve the therapeutic benefit of established regimens in early breast cancer Methods: From 1996 to 2002 a total of 1355 women with operable breast cancer (T>2cm) were randomized to adjuvant A (75 mg/m2 q21d x 4) followed by i.v. CMF (day 1&8 q28d x 4), or adjuvant A (60 mg/m2) and T (200 mg/m2 over 3 hrs q21d x 4) followed by CMF (AT→CMF), or AT→CMF as primary systemic therapy (PST). Results: Main patient characteristics (T-size, ER/PgR status, grade and age) were evenly distributed among arms. After median follow-up of 43 months, freedom from progression (FFP) was significantly better for women receiving adjuvant AT→CMF than A→CMF (HR 0.65, range 0.48–0.90, P=0.01). In a multivariate analysis, treatment inclusive of paclitaxel stood out as significantly associated with FFP (HR 0.66, P=0.012), together with clinical d...
537 Background: At the time the ECTO was designed in 1996, taxanes were only indicated for patien... more 537 Background: At the time the ECTO was designed in 1996, taxanes were only indicated for patients with metastatic breast cancer. However, paclitaxel and docetaxel were still to be tested in the adjuvant setting. In addition there was relatively scarce information on the comparative efficacy of neoadjuvant and adjuvant regimens. The ECTO trial was designed to evaluate the addition of paclitaxel to an anthracycline-based adjuvant regimen and to compare this combination with the same regimen given as primary systemic (neoadjuvant) therapy. Methods: A total of 1,355 women with operable breast cancer were randomized to one of three treatments: 1) surgery followed by adjuvant single agent doxorubicin (A) followed by CMF (arm A); 2) surgery followed by adjuvant paclitaxel plus doxorubicin (AT) followed by CMF (arm B); 3) AT followed by CMF followed by surgery (arm C). The two co-primary objectives were to assess the effects on freedom from progression (FFP) of: 1) the addition of paclitaxel to post-operative chemotherapy (arm B versus arm A); and 2) primary versus adjuvant chemotherapy (arm B versus arm C). Results: At 10 years, in the adjuvant setting FFP remained statistically significant in favor of AT followed by CMF (arm B, HR 0.77, P=0.045). Distant FFP was similarly improved but overall survival was not (HR 0.82, P=0.24). There was no significant difference in FFP when chemotherapy was given after surgery compared with the same regimen given before surgery (arm B vs arm C, HR 0.79, P=0.07). In the primary chemotherapy arm, patients who achieved a pathological complete remission (pCR) had improved distant FFP (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.001) compared to patients who did not achieve pCR. When given as primary systemic therapy, the paclitaxel-containing regimen allowed breast-sparing surgery in a significant percentage of patients, which did not translate in an increased risk of ipsilateral breast recurrence compared to the risk observed in patients in the adjuvant arms. Conclusions: Incorporating paclitaxel into anthracycline-based adjuvant therapy resulted in a significantly improved FFP and DFFP.
Trastuzumab treatment has significantly improved the prognosis of HER2-positive breast cancer pat... more Trastuzumab treatment has significantly improved the prognosis of HER2-positive breast cancer patients. Despite this, resistance to therapy still remains the main clinical challenge. In order to evaluate the implication of microRNAs in the trastuzumab response, we performed a microRNA array in parental and acquired trastuzumab-resistant HER2-positive breast cancer cell lines. Our results identified miR-146a-5p as the main dysregulated microRNA. Interestingly, high miR-146a-5p expression in primary tumor tissue significantly correlated with shorter disease-free survival in HER2-positive breast cancer patients. The gain- and loss-of-function of miR-146a-5p modulated the response to trastuzumab. Furthermore, the overexpression of miR-146a-5p increased migration and angiogenesis, and promoted cell cycle progression by reducing CDKN1A expression. Exosomes from trastuzumab-resistant cells showed a high level of miR-146a-5p expression compared with the parental cells. In addition, the co-c...
Anti-HER2 therapies have markedly improved prognosis of HER2-positive breast cancer. However, dif... more Anti-HER2 therapies have markedly improved prognosis of HER2-positive breast cancer. However, different mechanisms play a role in treatment resistance. Here, we identified AXL overexpression as an essential mechanism of trastuzumab resistance. AXL orchestrates epithelial-to-mesenchymal transition and heterodimerizes with HER2, leading to activation of PI3K/AKT and MAPK pathways in a ligand-independent manner. Genetic depletion and pharmacological inhibition of AXL restored trastuzumab response in vitro and in vivo. AXL inhibitor plus trastuzumab achieved complete regression in trastuzumab-resistant patient-derived xenograft models. Moreover, AXL expression in HER2-positive primary tumors was able to predict prognosis. Data from the PAMELA trial showed a change in AXL expression during neoadjuvant dual HER2 blockade, supporting its role in resistance. Therefore, our study highlights the importance of targeting AXL in combination with anti-HER2 drugs across HER2-amplified breast cance...
Novel antibody-drug conjugates against HER2 are showing high activity in HER2-negative breast can... more Novel antibody-drug conjugates against HER2 are showing high activity in HER2-negative breast cancer (BC) with low HER2 expression (i.e., 1+ or 2+ and lack of ERBB2 amplification). However, the clinical and molecular features of HER2-low BC are yet to be elucidated. Here, we collected retrospective clinicopathological and PAM50 data from 3,689 patients with HER2-negative disease and made the following observations. First, the proportion of HER2-low was higher in HR-positive disease (65.4%) than triple-negative BC (TNBC, 36.6%). Second, within HR-positive disease, ERBB2 and luminal-related genes were more expressed in HER2-low than HER2 0. In contrast, no gene was found differentially expressed in TNBC according to HER2 expression. Third, within HER2-low, ERBB2 levels were higher in HR-positive disease than TNBC. Fourth, HER2-low was not associated with overall survival in HR-positive disease and TNBC. Finally, the reproducibility of HER2-low among pathologists was suboptimal. This s...
532 Background: NOAH (NeOAdjuvant Herceptin) is a Phase III trial of neoadjuvant trastuzumab; H) ... more 532 Background: NOAH (NeOAdjuvant Herceptin) is a Phase III trial of neoadjuvant trastuzumab; H) in combination with chemotherapy in patients (pts) with HER2-positive locally advanced breast cancer (LABC). Methods: 228 pts with centrally confirmed HER2-positive (IHC 3+ or FISH+) LABC received 3 cycles of doxorubicin-paclitaxel (AT: A 60 mg/m2, T 150 mg/m2 q3w), 4 cycles of T (175 mg/m2 q3w) and 3 cycles of cyclophosphamide/methotrexate/5-fluorouracil (CMF: C 600 mg/m2, M 40 mg/m2, F 600 mg/m2 q4w) on days 1 and 8, with (n=115) or without (n=113) concomitant H (8 mg/kg loading dose then 6 mg/kg q3w for 1 year) before surgery. Pts with HER2- negative disease (IHC 0/1+; n=99) were treated in parallel with AT/T/CMF. The primary end point was event-free survival (EFS); secondary end points included overall response rate (ORR), pathological complete response (pCR) rate and safety. Results: Baseline characteristics were well balanced for randomised pts. Median tumour size was 5.5 cm (range...
Natural killer (NK) cells can orchestrate effective antitumor immunity. The presence of tumor-inf... more Natural killer (NK) cells can orchestrate effective antitumor immunity. The presence of tumor-infiltrating NK cells in diagnostic biopsies predicts pathologic complete response (pCR) to HER2-specific therapeutic antibodies in patients with primary breast cancer. Here, we analyzed whether diversity in circulating NK cells might influence tumor infiltration and HER2-specific therapeutic antibody efficacy. We found that numbers of circulating CD57+ NK cells inversely correlated with pCR to HER2-specific antibody treatment in patients with primary breast cancer independently of age, traditional clinicopathologic factors, and CD16A 158F/V genotype. This association was uncoupled from the expression of other NK-cell receptors, the presence of adaptive NK cells, or changes in major T-cell subsets, reminiscent of cytomegalovirus-induced immunomodulation. NK-cell activation against trastuzumab-coated HER2+ breast cancer cells was comparable in patients with high and low proportions of CD57+ ...
Purpose: This study aimed to identify biomarkers of resistance to endocrine therapy in estrogen r... more Purpose: This study aimed to identify biomarkers of resistance to endocrine therapy in estrogen receptor–positive (ER+) breast cancers treated with prolonged neoadjuvant letrozole.Experimental Design: We performed targeted DNA and RNA sequencing in 68 ER+ breast cancers from patients treated with preoperative letrozole (median, 7 months).Results: Twenty-four tumors (35%) exhibited a PEPI score ≥4 and/or recurred after a median of 58 months and were considered endocrine resistant. Integration of the 47 most upregulated genes (log FC > 1, FDR < 0.03) in letrozole-resistant tumors with transcription-binding data showed significant overlap with 20 E2F4-regulated genes (P = 2.56E−15). In patients treated with the CDK4/6 inhibitor palbociclib before surgery, treatment significantly decreased expression of 24 of the 47 most upregulated genes in letrozole-resistant tumors, including 18 of the 20 E2F4 target genes. In long-term estrogen-deprived ER+ breast cancer cells, palbociclib als...
BackgroundNeoadjuvant treatment is increasingly one of the preferred therapeutic options for earl... more BackgroundNeoadjuvant treatment is increasingly one of the preferred therapeutic options for early breast cancer and may have some unique outcomes, such as identifying predictive and prognostic factors of response or increasing the knowledge of individual tumor biology.DesignA panel of experts from different specialties reviewed published clinical studies on the neoadjuvant management of breast cancer. Recommendations were made that emphasized the clinical multidisciplinary management and the investigational leverage in early breast cancer.ResultsNeoadjuvant therapy has equivalent efficacy to adjuvant therapy, and it has some additional benefits that include increasing breast conservation, assessing tumor response, establishing prognosis based on the pathological response, and providing a “second opportunity” for nonresponding patients. Achieving pathological complete remission because of neoadjuvant therapy has been correlated with long-term clinical benefit, particularly in HER2-p...
Clinical cancer research : an official journal of the American Association for Cancer Research, Jan 6, 2018
We investigated the value of tumor-infiltrating NK cells (TI-NK) and HLA class I tumor expression... more We investigated the value of tumor-infiltrating NK cells (TI-NK) and HLA class I tumor expression as biomarkers of response to neoadjuvant anti-HER2 antibody-based treatment in breast cancer. TI-NK cells and HLA-I were determined by immunohistochemistry in pre-treatment tumor biopsies from two cohorts of HER2-positive breast cancer patients [discovery cohort (n=42) and validation cohort (n=71)]. TIL were scored according to international guidelines. Biomarker association with pathological complete response (pCR) and disease-free survival (DFS) was adjusted for prognostic factors. Gene set variation analysis was used for determining immune-cell populations concomitant to NK cell enrichment in HER2-positive tumors from the TCGA (n=190). TI-NK cells were significantly associated with pCR in the discovery cohort as well as in the validation cohort (p<0.0001), independently of clinicopathological factors. A ≥3 TI-NK cells/50xHPF cut off predicted pCR in the discovery and validation co...
554 Background: Gene expression profiling can be used to predict the likelihood of achieving a pa... more 554 Background: Gene expression profiling can be used to predict the likelihood of achieving a pathological complete response (pCR) to neoadjuvant chemotherapy (CT) in patients with primary breast cancer. We report a retrospective exploratory analysis to evaluate the utility of the PAM50 subtype assay in predicting pCR in women enrolled in the NeOAdjuvant Herceptin (NOAH) trial, which recruited 334 patients. METHODS Formalin-fixed paraffin-embedded core biopsies were collected from 156 patients randomized to receive doxorubicin/paclitaxel (AT) followed by CMF (Group 1; HER2-positive/negative), or the same regimen with trastuzumab (H) (AT+H→CMF+H; Group 2; HER2-positive). Total RNA was amplified, labeled, and hybridized onto Affymetrix 2.0 Plus microarrays. The PAM50 microarray-based assay was applied to 43 genes. The PAM50 Risk Of Relapse score based on subtype and Proliferation (RORP) was evaluated as a continuous variable and as low/medium/high-risk groups using previously reported cutoffs. pCR was defined by the absence of residual invasive tumor in the breast and axillary lymph nodes. Univariate logistic regression was used to evaluate the association of each signature with pCR. RESULTS Overall pCR rates were 21.5% (20/93) and 44.4% (28/63) in Groups 1 and 2, respectively (p<0.005). In Group 1, non-LumA tumors showed higher pCR rates than LumA tumors (27% vs. 0%, p<0.005). Low, medium and high RORP groups showed 0%, 19% and 37% pCR rates (p=0.02). In Group 2, all subtypes were identified and HER2-enriched tumors showed a non-significant increase in pCR rates compared to non-HER2-enriched tumors (53% vs. 34%). Low, medium and high RORP groups showed 17%, 36% and 75% pCR rates (p=0.01). In Group 2 the odds ratio for achieving a pCR in the high-risk RORP group was 15.0 compared to the low-risk group (p=0.029). CONCLUSIONS HER2-positive tumors predicted to have a high risk of relapse by the PAM50 assay benefited substantially from H-based CT. Similar findings are observed in HER2-positive/negative tumors treated with CT only. The role of PAM50 and/or other genomics assays in predicting pCR and survival during treatment with anti-HER2 agents warrants further investigation.
529 Background: HER2-positive/ER+ and HER2-positive/ER- breast cancers have different patterns of... more 529 Background: HER2-positive/ER+ and HER2-positive/ER- breast cancers have different patterns of gene expression and intracellular pathway activation. We assessed the role of selected biomarkers in predicting the likelihood of achieving a pathological complete response (pCR) in the NeOAdjuvant Herceptin (NOAH) trial, which recruited 334 patients. METHODS Formalin-fixed paraffin-embedded core biopsies were collected from 114 women randomized to receive trastuzumab (H) with doxorubicin/paclitaxel followed by CMF (Arm CTH) or chemotherapy alone (Arm CT). Expression of the following genes and metagenes was profiled on Affymetrix 2.0 Plus: MYC, PIK3CA, PIK3R1, PTEN, amphiregulin (AREG), epiregulin, 8q24 and 8q22 amplicons, plasma cell (PC) and IGF metagenes. ER status was based on ESR1 gene expression. pCR was defined as the absence of residual invasive tumor in the breast and axillary lymph nodes. Biomarkers were assessed as continuous variables by univariate logistic regression analysis. RESULTS 80 samples (70%) were ER- and 34 (30%) were ER+. In the CT arm AREG (p=0.03) and 8q22 (p=0.06) were associated with lower pCR rates among ER- samples, while PTEN (p=0.01) was associated with higher pCR rates. Among ER- samples in the CTH arm, AREG (p=0.04) and IGF (p=0.007) were associated with resistance, while PC showed a trend for higher pCR rate (p=0.079). There were significant interactions between the treatment arms and 8q22 (p=0.03) and PC (p=0.04). In the high tertile of 8q22 the pCR rates were 0% (0/12 Arm CT) and 67% (10/15 Arm CTH) while for PC the pCR rates were 13% (2/15 Arm CT) and 83% (10/12 Arm CTH). There were no statistically significant associations for ER+ tumors, but a significant interaction for the likelihood of pCR and ER status was observed for PTEN (p=0.006) and PC (p=0.05) in the CT Arm and for AREG (p=0.03) in the CTH Arm. CONCLUSIONS High expression of the PC metagene and the 8q22 amplicon, and low expression of the IGF metagene, were associated with the increased pCR rate reported with the addition of H to CT in HER2-positive/ER- tumors. The concept that HER2-positive/ER+ and HER2-positive/ER- tumors are driven by different biologic pathways warrants further investigation.
Clinical cancer research : an official journal of the American Association for Cancer Research, 2002
The full-length receptor p185HER-2 undergoes a metalloprotease-dependent cleavage producing a mem... more The full-length receptor p185HER-2 undergoes a metalloprotease-dependent cleavage producing a membrane-associated fragment (p95HER-2) in cultured breast cancer cells. P95HER-2 has potentially enhanced signaling activity, but its expression and role in human breast cancer is poorly characterized. The purpose of this project was to characterize the expression of p95HER-2 in primary breast cancers and nodal metastasis, and to study association with clinicopathological factors. P95HER-2 and p185HER-2 were examined in 337 primary breast tumors and 81 metastatic lymph nodes by Western blot analysis, and tested for associations with other clinicopathological factors. P95HER-2 was present in 20.9% of primary tumors from node-negative patients, in 29.1% from patients with one to three metastatic nodes, and in 36.7% from patients with four or more metastatic nodes (P = 0.027). Whereas p185HER-2 overexpression was unrelated to nodal disease (P = 0.63), the odds of lymph node metastasis were en...
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Papers by Ana Lluch