bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract P4-14-16: T-cell clonality increases after neoadjuvant treatment with trastuzumab and pertuzumab
Authors: Howie LJ , Marcom PK , Topping DL , Force J , Emerson R , Bhavsar NA , Abbott SE , Parks M , Robins HS , Blackwell KL 
Published: May 2016

Abstract
Background: Baseline TIL density and T-cell clonality predict pathological complete response (pCR) to neoadjuvant (NA) trastuzumab (TRAS). Preclinical data suggests that a major mechanism of TRAS is enhancement of antibody-dependent cytoxicity (ADCC) and T-cell mediated tumor kill and that the combination of TRAS with pertuzumab (PERT) may enhance ADCC. This study quantifies both TIL density and T-cell clonality in both pre and post-NA samples from patients receiving NA TRAS +/- PERT.
Methods: Sequential subjects treated with NA docetaxel, carboplatin and TRAS +/- PERT were identified and pre-treatment (n=39) and post-treatment specimens (n=44, 36 pairs of pre and post-treatment; 19 with pCR) were sequenced using the ImmunoSEQ platform (Adaptive Bioscience, Seattle, WA). T-cell density and dominant T-cell clonality were measured by TCR sequencing and then associated with pCR using a two-tailed Mann-Whitney U test comparing patients with pCR versus those without. Paired values were compared using the sign test to evaluate change in pre and post-treatment T-cell density and clonality. TIL density was performed using standard IHC methodology.
Results: There was a trend towards increased T-cell clonality after treatment (p=0.08). T-cell clonality post-treatment was significantly increased compared to pre-treatment in those samples where post-treatment tumor was available (p=0.01). There was no difference in T-cell clonality between pre- and post-treatment in normal tissue from pts achieving a pCR (p=1.00). There was no difference in post-treatment T cell clonality when treated with TRAS vs. TRAS + PERT (p=0.19). No differences in the % of pretreatment TILs or T-cell clonality were seen in those who obtained a pCR vs. those who did not (p=0.39 and p=0.85). In addition, no differences in T-cell clonality were seen based on ER status in either pre-treatment or post-treatment samples (p=0.40/0.48)
Conclusions: After treatment with TRAS +/- PERT, there was an increase in T-cell clonality suggesting an enhanced immune response as a mechanism of action for HER2 targeting antibodies. This analysis is currently limited by a small sample size and high rate of pCR. We plan to include additional paired samples which may help to better demonstrate the increased immune response that occurs with HER-2 targeting antibodies.
Citation Format: Howie LJ, Marcom PK, Topping DL, Force J, Emerson R, Bhavsar NA, Abbott SE, Parks M, Robins HS, Blackwell KL. T-cell clonality increases after neoadjuvant treatment with trastuzumab and pertuzumab. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P4-14-16.
Abstract P5-01-01: Real-time imaging of lymph node metastasis in response to systemic ezrin inhibitor treatment in breast cancer
Authors: Ghaffari A , Hoskin V , Mullins G , Greer P , Kiefer F , Madarnas Y , SenGupta S , Elliott B 
Published: May 2016

Abstract
Lymph node (LN) metastasis is a key driver of recurrence and survival in breast cancer (BC) patients. However, the mechanisms of metastatic dissemination of tumour cells from LNs to distant sites and their predictors of response to systemic therapy remain poorly understood, mainly due to a lack of non-invasive in vivo imaging models. We have recently described ezrin, a pro-metastatic crosslinker protein, as a regulator of tumour lymphangiogenesis and metastasis in BC (Breast Cancer Res. 2014; 16(5): 438). Furthermore, we demonstrated significant association of high ezrin expression with lymphovascular invasion in a cohort (n=63) of premenopausal patients with invasive BC (p =0.024). These findings prompted us to examine the role of ezrin in migration and invasion of metastatic tumour cells in LNs and their response to ezrin-targeted therapy. Using a locally accrued LN positive patient cohort (n=94), we demonstrated a significant association between high ezrin levels and reduced recurrence-free survival (univariate Log-rank test, p=0.033), suggesting that ezrin is a potential predictor of relapse in LN positive BC. To address the mechanistic role of ezrin in LN metastasis, we developed a novel intravital imaging model using a lymphatic reporter transgenic mouse (B6-prox1-mOrange2-pA-BAC) to examine the response of tumour-draining LN to anti-ezrin systemic therapy in real time. Next, we tested the effects of a small molecule ezrin inhibitor (NSC668394) in vitro and observed significant suppression of ezrin activation (p-T567) and cancer cell invasive phenotype. Intravital imaging of inguinal LN metastases, derived from subcutaneously implanted breast adenocarcinoma E0771-LMV (lung metastatic variant) cells, demonstrated significant reduction in mobility and invasiveness (Mann Whitney, p&lt;0.0001) of metastatic cells following systemic treatment with NSC668394 (0.5 mg/kg at 24h and 8h prior to imaging). Interestingly, LN metastases engagement by host T cell (CD3+) was notably increased, whereas T cell mobility was not affected by ezrin inhibition. Our findings present a novel non-invasive imaging model to study the LN metastasis response to anti-cancer therapy in real time, and provide new insight into the role of ezrin as a potential anti-metastatic target in BC.
(Supported by CRS, CIHR, CBCF, BCAK, Queen's SRC).
Citation Format: Ghaffari A, Hoskin V, Mullins G, Greer P, Kiefer F, Madarnas Y, SenGupta S, Elliott B. Real-time imaging of lymph node metastasis in response to systemic ezrin inhibitor treatment in breast cancer. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P5-01-01.
Abstract P5-03-14: MLN0128 regulates survival signaling by AKT and its downstream effectors in HER2+ breast cancer model
Authors: De PK , Carlson JH , Sun Y , Lin X , Williams C , Dey N , Leyland-Jones BR 
Published: May 2016

Abstract
Evading apoptosis is considered to be a hallmark of cancers including breast cancer, since mutations in apoptotic regulators invariably accompany tumorigenesis. Chemotherapeutic agents induce apoptosis, and hence disruption of apoptosis during tumor progression may promote drug resistance. AKT is an apoptotic regulator that is activated in HER2+ breast tumor cells and promotes anti-HER2 therapy resistance in vitro. Nevertheless, how mTORC1/C2-AKT signaling disables apoptosis and its contribution to clinical drug resistance are not clear yet. Using HER2 amplified breast cancer cells [BT474 (HER2+/Trastuzumab-sensitive), BT474HerR (HER2+/Trastuzumab-resistant), HCC1954 and MDA-MB453 (both are HER2+/PIK3CA kinase domain mutated)], we show that mTORC1/C2 inhibitor; MLN0128 abrogates AKT (Ser473), Survivin and controls its downstream effectors of apoptotic signaling molecules (e.g. cleaved Caspase 3/9, cleaved PARP, MCL and BIM). MLN0128 also induces annexinV positive cells and regulates cellular proliferation (ON-TOP 3D colony formation and real-time proliferation assay). Additionally, increased cleaved Caspase 3 and decreased MCL1 expression were also observed following MLN0128 treatment in HER2+ xenograft model along with tumor growth inhibition. Our studies provide strong experimental evidence that high apoptotic signaling –specifically reduced MCL1 and increased cleaved-CASPASE3 expression expedite the response of targeted therapy that directly inhibits mTORC1/C2-AKT signaling.
Citation Format: De PK, Carlson JH, Sun Y, Lin X, Williams C, Dey N, Leyland-Jones BR. MLN0128 regulates survival signaling by AKT and its downstream effectors in HER2+ breast cancer model. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P5-03-14.
Abstract P5-04-19: JAK2 copy number and targeted JAK2 inhibition of TNBC cell lines
Authors: Chen M , Andreozzi M , Gonzalez-Malerva L , Eaton S , Pockaj B , Barrett MT , Anderson KS 
Published: May 2016

Abstract
Background A subset of triple negative breast cancers (TNBC) has somatic amplification of chromosome 9p24.1, encoding PD-L1, PD-L2, and JAK2 (the "PDJ" amplicon). The JAK/STAT pathway promotes malignant transformation and proliferation, and the JAK1/JAK2 inhibitor ruxolitinib is being evaluated in early phase clinical trials in breast cancer. This study was designed to measure the frequency of PDJ copy number gain in existing TNBC cell lines, and determine the impact of targeted JAK2 inhibition on cellular proliferation.
Methods: Copy number alterations were measured in nine TNBC cell lines (MDA-MB-453, MDA-MB-436, BT549, MDA-MB231, MFM-223, MDA-MB-468, H5578T, MDA-MB-157, HCC1937) by array comparative genomic hybridization (aCGH). JAK2 and pSTAT3 expression was measured by immunoblot in four of the TNBC lines (HCC1937, MDA-MB-468, MDA-MB-231, MDA-MB-436) and compared with MCF10A, SK-BR3, and T47D. To selectively inhibit JAK2, lentiviral vectors encoding five different shRNA targeting JAK2 were generated. Ruxolitinib treatment was performed at 25 uM for 48 hrs and 72 hrs. Cell proliferation was measured by CellTiter-Glo.
Results: All TNBC cell lines had an elevated level of baseline protein expression of JAK2 and pSTAT3 compared to SK-BR3, T47D, and MCF10A. Of the nine TNBC cell lines, only MDA-MB-436 and BT549 had copy number gain of 9p24.1 (log2ratio&lt;1). Four independent shRNA efficiently silenced over 90% of JAK2 expression in MDA-MB-231. The inhibition of JAK2 in MDA-MB-231 was associated with a decrease in both JAK2 and pSTAT3 protein expression and reduction in cell proliferation compared to control shRNA after 96 hrs (p&lt;0.0001). Inhibition of proliferation was also observed for both MDA-MB-231 and MDA-MB-436 after ruxolitinib treatment.
Conclusion These results demonstrate that existing TNBC cell lines have varying copy number alterations of chromosome 9p24.1 encoding JAK2, and targeted JAK2 inhibition in TNBC inhibits cell proliferation.
Citation Format: Chen M, Andreozzi M, Gonzalez-Malerva L, Eaton S, Pockaj B, Barrett MT, Anderson KS. JAK2 copy number and targeted JAK2 inhibition of TNBC cell lines. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P5-04-19.
Abstract P5-05-03: Targeting hypoxia-induced cancer cell metabolism in breast cancer
Authors: Cavazos DA , Gruslova A , Garcia M , Brenner AJ 
Published: May 2016

Abstract
Rational: Hypoxia contributes to high tumor grade in human breast cancer and is associated with resistance to radiation and chemotherapy. Hypoxia is generally encountered in perinecrotic regions throughout the heterogeneous tumor microenvironment and may be due to rapid tumor grow, insufficient neovascularization or as a result of antiangiogenic and chemoradiation therapies which cause acute vascular thrombosis. Cancer stem-like cells residing within these hypoxic zones compensate for the low concentration of oxygen through alterations in metabolism that favor aerobic glycolysis and de novo fatty acid synthesis. These metabolic derangements are known to enhance proliferation and survival, although the contributing molecular mechanisms are not fully understood. Here we evaluate a panel of FDA approved azole drugs for alternative use as glycolytic inhibitors with and without the fatty acid synthase (FASN) inhibitor, TBV-3166. TVB-3166 is currently in phase II clinical trials and is the first FASNi approved for human testing. METHODS: IC50 values of single or combined agents were determined by MTT under conditions of normoxia and hypoxia using sister plates of estrogen receptor positive or triple negative breast cancer cells, MCF-7, SKBR3 and MDA-MB-231. RESULTS: Because the combination of Itraconazole and TVB-3166 displayed a synergistic enhancement in cell death at a low nanomolar concentrations in MCF-7 cells, we next assessed the efficacy of these drugs in an in vivo xenograft model of breast cancer. Here, 3.6 million MCF-7 cells were implanted into the right flanks of (n=16) athymic nude mice, which were then randomized by tumor volume into 4 groups. Itraconazole and TVB-3166 were administered at 60 mg/kg daily by oral gavage as single agents or combined throughout the duration of the study. After three weeks, mice treated with combined inhibitors exhibited best response with a statistically significant decrease in tumor volume compared to the vehicle control group. CONCLUSION: Given the significant tumor response to the combination of metabolic inhibitors, further studies that evaluate the radiosensitizing effects under conditions of hypoxia are warranted. By targeting hypoxia-induced metabolic pathways during radiotherapy, we may enhance cancer stem-like cell death to dramatically improve patient outcome.
Citation Format: Cavazos DA, Gruslova A, Garcia M, Brenner AJ. Targeting hypoxia-induced cancer cell metabolism in breast cancer. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P5-05-03.
Abstract P5-07-08: The clinical utility of alternative chromosome 17 probes in equivocal HER2 results
Authors: Sneige N , Gong Y , Multani AS , Ibrahim NK 
Published: May 2016

Abstract
Background
Acurrate assessment of HER2 status in patients with breast cancer is critical in selecting patients for targeted therapy. In 2013, the American Society of Clinical Oncology-/College of American Pathologists (ASCO/CAP) released new guidelines for HER2 testing in an attempt to more acurately identify all breast cancer patients who are eligible for HER2 targeted therapy; these guidelines included new cutoff points for HER2/CEP17 ratio by in situ hybridization (ISH) or average HER2 copy number/cell and recommended the use of a reflex test with alternative ISH chromosome 17 probe for resolving cases with equivocal HER2 ISH results. We sought to determine the clinical utility of alternative chromosome 17 probes in such equivocal cases.
Patients and Methods
Our institution's database of HER2 dual- probe ISH results was searched for cases of invasive breast cancer with a HER2/CEP17 ratio of &lt;2 and an average HER2 copy number of &gt;4 and &lt;6 signals/cell (i.e., cases meeting the new ASCO/CAP definition of equivocal). Of these, 30 consecutive cases with available corresponding sections were selected for additional chromosome 17 studies using probes for Smith-Magenis syndrome (SMS) and retinoic acid receptor alpha (RARA) genes. A eusomic copy number exhibited in one or both of these loci was used to calculate a revised HER2/chromosome-17 ratio by using the eusomic gene locus as the reference.
Results
Of 5489 cases of invasive breast cancer tested by dual-probe ISH for HER2 gene status, 316 (5.7%) cases were found to have equivocal results according to the 2013 ASCO/CAP guidelines. Further analysis of the 30 equivocal consecutive cases using the alternative chromosome 17 probes (SMS/RARA), 17 cases were upgraded from equivocal to positive HER2 status, and 13 cases remained unchanged.
Conclusion
The use of alternative chromosome 17 probes effectively determines true HER2 status in most equivocal HER2 ISH cases. Equivocal results are common; therefore, we advocate testing for alternative chromosome 17 genes to better categorize equivocal cases under the new ASCO/CAP guidelines. Additional analysis of the remaining equivocal cases and their relation to disease progression is underway and will be presented at the meeting.
Citation Format: Sneige N, Gong Y, Multani AS, Ibrahim NK. The clinical utility of alternative chromosome 17 probes in equivocal HER2 results. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P5-07-08.
Abstract P5-13-06: Concurrent gonadotropin-releasing hormone (GnRH) agonist administration with chemotherapy improves neoadjuvant chemotherapy responses in young premenopausal breast cancer patients
Authors: Yoon TI , Kim HJ , Yu JH , Sohn G , Ko BS , Lee JW , Son BH , Ahn SH 
Published: May 2016

Abstract
Background
Gonadotropin-releasing hormone (GnRH) agonist therapy for ovarian function preservation shows promising results. This study aimed to determine the oncologic efficacy of GnRH agonist treatment concurrent with chemotherapy in a neoadjuvant setting.
Patients and Methods
A retrospective analysis was performed on 332 cases of invasive breast cancer in patients who were &lt;40 years old at diagnosis and received GnRH agonists concurrent with neoadjuvant chemotherapy (GnRH agonist group) or neoadjuvant chemotherapy alone (neochemotherapy-alone group) at Asan Medical Center from December 2010 to September 2014. Pathologic complete response rates (pCR) and Ki-67 changes were evaluated between the two groups. For hormone receptor (HR)-positive tumors, the clinical response and preoperative endocrine prognostic index (PEPI) score also were evaluated.
Results
The median age was 32 ± 3.9 and 36 ± 3.0 years old in the GnRH agonist group and neochemotherapy-alone group, respectively (P &lt; .001). Adjusted for tumor size, grade, lymph node metastasis, HR status, and chemotherapy regimen, the GnRH agonist group exhibited a higher pCR rate with an odds ratio (OR) of 2.98 (95% CI, 1.37–6.34) and more decreased Ki-67 expression during treatment (P = 0.05) than the neochemotherapy-alone group. In HR-negative tumors, the GnRH agonist group showed a higher pCR rate (multivariate OR = 3.50; 95% CI, 1.37–8.95) and more decreased Ki-67 expression (P = 0.047). In HR-positive breast cancer, the pCR rate, change in Ki-67 index, and clinical response were higher and preoperative prognostic index (PEPI) scores were lower in the GnRH agonist group, but not significant between the two treatment groups.
Conclusion
Concurrent administration of GnRH agonists during neoadjuvant chemotherapy improved pCR rates and suppressed Ki-67 expression especially in HR-negative tumors.
Citation Format: Yoon TI, Kim HJ, Yu JH, Sohn G, Ko BS, Lee JW, Son BH, Ahn SH. Concurrent gonadotropin-releasing hormone (GnRH) agonist administration with chemotherapy improves neoadjuvant chemotherapy responses in young premenopausal breast cancer patients. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P5-13-06.
Abstract P6-03-15: Inhibition of breast cancer tumor cell lines by simvastatin is dependent on specific p53 mutation
Authors: Ferdosi SR , Grewal H , Gonzalez-Malerva L , Eaton S , Briones N , LaBaer J , Anderson KS 
Published: May 2016

Abstract
Background: Simvastatin is an HMG-CoA reductase inhibitor widely used to treat cardiovascular diseases. In retrospective studies, statin treatment has been associated with a modest decrease in overall cancer incidence, including breast cancer (10-15%). Simvastatin Inhibited the PI3K/Akt/mTOR pathway in a pilot window-of-opportunity trial in neoadjuvant breast cancer. Mutation in TP53 is correlated with elevated expression of genes regulating cholesterol biosynthesis and sensitivity to statin treatment in vitro. We hypothesized that specific mutations in TP53 would be associated with differential sensitivity to simvastatin.
Method: We generated MCF10A stably transduced cell lines over-expressing ten frequent TP53 missense point mutations. We assessed the impact of TP53 mutation on growth inhibition induced by simvastatin treatment (range of 1-30 µM for 96 hrs). In parallel, induction of apoptosis was measured by caspase3 reporter assay. Illumina 4-plexed 1x50bp RNA sequencing was performed on cells with R273H, G245S, R248Q, Y234C and wt TP53 before and after exposure to 2.5 µM simvastatin.
Results: We confirmed that mutation in TP53 was markedly associated with growth inhibition by simvastatin treatment in vitro. We now demonstrate that TP53 mutation in R273C, G245S and R273H are highly sensitive to simvastatin with IC50 values ≤ 2.1 µM. In contrast, TP53 mutations in Y234C and R248Q were approximately five-fold more resistant. The resistant mutations were similar to MCF10A overexpressing wild type TP53. Growth inhibition was correlated with induction of apoptosis, and could be rescued by addition of farnesylpyrpphosphate and geranylgeranylpyrophosphate. HMGCR upregulation was observed across all treated cell lines. RNA-seq analysis confirms down regulation of the PIK3CA, PIK3CB, and Akt in the sensitive cell lines, but paradoxical upregulation in the resistant cells. Further gene expression analysis will be presented.
Conclusion: Specific TP53 mutation status may impact sensitivity of breast cancer to simvastatin treatment.
Citation Format: Ferdosi SR, Grewal H, Gonzalez-Malerva L, Eaton S, Briones N, LaBaer J, Anderson KS. Inhibition of breast cancer tumor cell lines by simvastatin is dependent on specific p53 mutation. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P6-03-15.
Abstract P6-03-18: Outcome of FoundationOne testing in metastatic breast cancer therapy: A single center experience
Authors: Yuan Y , Mortimer JE 
Published: May 2016

Abstract
Background: Next generation sequencing (NGS) has made genomic, mutation-driven, cancer medicine feasible. However, few studies have reported the importance of NGS in identifying mutation-driven treatment or whether such therapies are effective in women with metastatic breast cancer (MBC). We reviewed the impact of NGS in women with MBC at our institution.
Methods: Among the medical oncologists who treat metastatic breast cancers at the City of Hope Comprehensive Cancer Center, we identified 21 patients with HER2 negative cancers, whose tumors were submitted for FoundationOne genomic profiling. We report the individual mutation profile, potential therapeutic options, and treatment outcome.
Result: Of the 21 patients (pts), 13 had triple negative breast cancer (TNBC) and 12 of 13 (92%) had actionable mutation(s). Mutation profiles have been heterogeneous. Loss or mutations of TP53 were the most common genomic alteration in TNBC (12 of 13). Four pts received targeted therapies (everolimus in 3, pazopanib in 1) and all experienced at least partial clinical response. An additional 3 pts were treated with directed therapy but are too early to evaluate, 4 pts were not treated because of disease progression or clinical deterioration, and 1 pt was lost to follow-up. All 8 pts with ER+ disease had actionable mutations identified and 7 (88%) received targeted therapy (pazopanib in1, everolimus in 3, palbociclib in 3). All experienced at least partial clinical response; 1 was too ill for treatment. The most common genomic alterations are: PIK3CA mutation, PTEN loss, P53 mutation, FGFR mutation, and ZNF mutation.
Conclusion: Targeted genomic sequencing through FoundationOne can identify mutations that are amenable to treatment with agents that are not generally approved for use in breast cancer. In order to test the benefit of target-directed therapy, tumor should be submitted for NGS while patients have a good performance status.
Citation Format: Yuan Y, Mortimer JE. Outcome of FoundationOne testing in metastatic breast cancer therapy: A single center experience. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P6-03-18.