bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P2-01-22: KLF4 improve prognosis of triple negative breast cancer by suppression of epitherial mesenchymal transition
Authors: Nagata T , Sekine S , Arai M , Fujii T
Published: May 2019

Background

Triple negative breast cancer (TNBC) is highly malignant and prone to metastasis and relapse, and therefore has poorer prognosis than other sub-types. The mechanism of higher malignancy of TNBC has not been sufficiently elucidated. KLF4is reported to be a transcription factor that is associated with both tumor suppression and oncogenesis. We have reported that breast cancer patients with strong expression of KLF4 had better prognosis, especially in TNBC patients. And here we report that KLF4 negatively regulates the metastasis and growth of TNBC.

Methods

We assessed the expression levels of KLF4 in 84 patients with TNBC by immunohistochemical staining and studied the patterns of metastasis/recurrence clinicopathologically. The overall survival (OS) rate and the disease free survival (DFS) rate after surgery was calculated by Kaplan-Maier method. In addition, circulating tumor cells (CTCs) in the peripheral blood of TNBC patients were identified and compared with primary lesions in terms of KLF4 expression. Moreover, the expression of KLF4 was inhibited by transfecting cultured TNBC cells (MDA-MB231) with the small interfering RNA (siRNA) of KLF4 to analyze the effects of KLF4 on cell proliferation and epithelial-mesenchymal transition (EMT)-like changes. For the proliferation assay, measurements were made by MTT assays. Cell migration and invasion assays of KLF4 suppressed TNBC cells were also examined. Total RNA was extracted from these cells, cDNA was synthesized, and used for the quantitative polymerase chain reaction (qPCR) analysis.

Results

In the 84 TNBC patients, higher KLF4 expression was associated with significantly better OS and DFS. An analysis of KLF4 expression in CTCs of the TNBC patients showed that KLF4 expression was lower in CTCs than in primary cancer lesions. TNBC cells (MDA-MB231) that were transfected the KLF4 siRNA exhibited a greater ability to growth than controls. These cells also underwent EMT-like changes with reduced expression of epithelial factors such as E-cadherin. Treating these TNBC cells with eribulin resulted a reduction of the expression of stem cell/EMT markers.

Conclusion

TNBC patients with reduced KLF4 expression had poor outcomes. The results of our experiments suggest the expression of KLF4 is one of the important factors that inhibit the EMT and growth of TNBC.

Citation Format: Nagata T, Sekine S, Arai M, Fujii T. KLF4 improve prognosis of triple negative breast cancer by suppression of epitherial mesenchymal transition [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-01-22.
Abstract P2-02-03: FASN inhibition as a potential treatment for therapy of endocrine resistant breast cancer
Authors: Gruslova A , Sareddy GR , Vadlamudi RK , Viswanadhapalli S , Brenner A
Published: May 2019

INTRODUCTION: Fatty acid synthase (FASN) is a key enzyme in tumor cell biology controlling endogenous lipid biosynthesis. It is overexpressed in a biologically aggressive subset of tumors, including breast carcinoma. We previously reported prolonged stabilization of disease with TVB-2640 in patients with advanced metastatic breast cancer, including some endocrine resistant ER+ tumors. Using in vitro and in vivo models, we assessed the role of FASN inhibition by TVB-3166 (preclinical version of TVB-2640) for treatment of endocrine resistant breast cancer. METHODS: Breast tumor cells were incubated with TVB-3166 (200nM), imaged and analyzed by automated Live-Cell analysis system (IncuCyte). For tumor growth inhibition, cells (2X106)were subcutaneously injected into SCID mice implanted with estrogen pellets. Once tumors were measurable, mice were divided into treatment groups: tamoxifen (4mg/kg), TVB-3166 (60mg/kg) and the combination. Patient tumor explants were incubated for 72h on gelatin sponges in culture medium in the absence or presence of 200nM TVB-3166. Tissue were fixed in 10% formalin and processed into paraffin blocks. Sections were stained with H&E, ERα and Ki67. RESULTS: The effectiveness of FASN inhibition on the growth of tumor cells has been confirmed in a number of breast cancer cell lines such as MCF7, ZR75, MDA-MB-231 and others. TVB-3166 leads to a marked inhibition of growth in tamoxifen resistant (TamR) cells, which 15% greater than in the parent line. IHC and Western blot showed FASN inhibition leads to significantly reduction of ERα levels. Immunofluorescent confocal microscopy showed inhibition of FASN by TVB-3166 alters subcellular localization of ERα. TVB-3166 was able to significantly inhibit tamoxifen resistant breast tumor growth in mice (p<0.05). Additionally, TVB-3166 treatment of primary tumor explants decreased their proliferation (Ki67) compared to untreated controls (21% vs 38%, p<0.01). CONCLUSION: Our preclinical data provide evidence that FASN inhibition by TVB-3166 presents a promising therapeutic strategy for treating of endocrine resistant breast cancer. RNA sequencing of tumor explants is being performed to evaluate FASN inhibition impact on canonical and non-canonical ERα signaling pathways.

Citation Format: Gruslova A, Sareddy GR, Vadlamudi RK, Viswanadhapalli S, Brenner A. FASN inhibition as a potential treatment for therapy of endocrine resistant breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-02-03.
Abstract P2-02-09: Not presented
Authors: An J , Yang J , Lee W-H , Kim JB , Kim J-k , Kim H , Paek SH , Lee JW , Woo J , Kwon H , Lim W , Paik NS , Kim Y-K , Moon B-I
Published: May 2019

This abstract was not presented at the conference.

Citation Format: An J, Yang J, Lee W-H, Kim JB, Kim J-k, Kim H, Paek SH, Lee JW, Woo J, Kwon H, Lim W, Paik NS, Kim Y-K, Moon B-I. Not presented [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-02-09.
Abstract P2-02-11: Combinational treatment of biguanides and fatty acid β-oxidation inhibitor in triple-negative breast cancers
Authors: Park JH , Jung KH , Vithayathil S , Jia D , Kaipparettu BA
Published: May 2019

Among breast cancers (BCs), the driver pathways and therapeutic targets are still poorly understood for triple negative (TN) BCs. Advances in cancer metabolism research over the last decade have enhanced our understanding on metabolic reprogramming in cancer therapy. We have previously shown that metabolic reprogramming to fatty acid β-oxidation (FAO) is a major energy pathway in metastatic TNBC. Moreover, we reported that FAO regulates c-Src, one of the frequently upregulated oncopathways in TNBC via autophosphorylation of Src at Y419. Since FAO inhibitors alone cannot effectively control the tumor progression in TNBC, suitable combination therapies with other metabolic targets are necessary. Recently increasing evidences show that anti-diabetic biguanides have attractive anticancer effect in various cancer types including BC. However, its significance as an anticancer drug is not well established due to parallel metabolic pathways that support tumor growth.

Phenformin, a biguanide derivative similar to metformin, has a greater potency than metformin. Like metformin, phenformin also inhibits mitochondrial electron transport chain (ETC) through complex I inhibition. In addition, biguanides lead to the activation of AMPK, which plays a key role in insulin signaling and energy sensing. Importantly, AMPK is an upstream regulator of FAO pathway because it can phosphorylate ACC to activate FAO. Considering the dependency of TNBC to FAO, we evaluated the therapeutic significance of the combination of biguanides(ETC inhibitors) and FAO inhibitors in TNBC progression and metastasis. We hypothesize that blocking both 'arms' of the pathway can provide more pronounced and durable responses in TNBCs. Our different in vitro and in vivo studies using TNBC cell line and PDX models suggest that the combination of both inhibitors can provide better therapeutic significance in metastatic TNBCs. This is a rationale and cost-effective metabolic approach to manage the currently non-targetable metastatic TNBCs. Further investigation into the clinical effectiveness of this combination may provide better treatment opportunities for TNBC patients.

Citation Format: Park JH, Jung KH, Vithayathil S, Jia D, Kaipparettu BA. Combinational treatment of biguanides and fatty acid β-oxidation inhibitor in triple-negative breast cancers [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-02-11.
Abstract P2-02-13: Computational metabolism modeling predicts risk of relapse in breast cancer patients
Authors: Zamora P , Trilla-Fuertes L , Gámez-Pozo A , Prado-Vázquez G , Zapater-Moros A , Ferrer-Gómez M , Díaz- Almirón M , López Vacas R , Espinosa E , Fresno Vara JA
Published: May 2019

Introduction

Breast cancer is one of the most prevalent cancers in the world. In previous works we observed differences in glucose metabolism between breast cancer subtypes, suggesting that metabolism plays an important role in this disease. Flux Balance Analysis (FBA) is widely used to study metabolic networks, allowing predicting growth rates or the rate of production of a given metabolite.

Material and methods

Proteomics data from 96 breast cancer tumors were obtained applying a high-throughput proteomics approach to routinely archive formalin-fixed, paraffin-embedded tumor tissue. Proteomics tumor data were analyzed using the human metabolic reconstruction Recon2 and FBA. The tumor growth rate for each tumor was calculated. In order to analyze fluxes from the different metabolic pathways, flux activities were calculated as the sum of the fluxes of each reaction in each pathway defined in the Recon2. Then, flux activities were used to build prognostic models.

Results and discussion

Using the results obtained from FBA in the proteomics dataset, flux activities were calculated for each pathway. Employing these flux activities, a prognostic signature was built. Flux activities of vitamin A, tetrahydrobiopterin metabolism and beta-alanine metabolism pathways split our population into a low and a high risk group (p=0.044).

Conclusion

Vitamine A, beta-alanine and tetrahydrobiopterin metabolism flux activities could be used to predict relapse risk. Flux activities is a method proposed in a previous work to study response against drugs that now also demonstrated its utility in summarizing FBA data and is associated with prognosis.

Citation Format: Zamora P, Trilla-Fuertes L, Gámez-Pozo A, Prado-Vázquez G, Zapater-Moros A, Ferrer-Gómez M, Díaz- Almirón M, López Vacas R, Espinosa E, Fresno Vara JA. Computational metabolism modeling predicts risk of relapse in breast cancer patients [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-02-13.
Abstract P2-03-04: A novel druggable target upstream of Notch: MEK5/ERK5 signaling regulates Jagged-1 and Notch1 expression in triple negative breast cancer stem cells
Authors: Ucar DA , Matossian MD , Hoang-Barnes VT , Hossain FM , Gupta M , Burks HE , Wright TD , Cavanaugh J , Flaherty P , Burow ME , Miele L
Published: May 2019

Triple negative breast cancer (TNBC) is a molecularly heterogeneous, clinically aggressive disease group that is highly prevalent among African-Americans and younger patients. Standard chemo/radio therapy often produces clinical responses, but recurrence and metastasis are unfortunately common. Metastatic disease is generally incurable. Chemo/radiotherapy has been shown to induce EMT and enrich a chemo-resistant cancer stem-like cell (CSC) population in TNBC. CSCs are thought to drive disease recurrence. Notch signaling is critical for maintenance of TNBC CSC. Expression of Notch1 and its ligand Jagged1 are correlated with poor prognosis. Efforts to pharmacologically target Notch with Gamma Secretase Inhibitors (GSIs) have been impaired by the systemic toxicity of the GSIs, and by the fact that Notch1 also plays a key role in anti-tumor adaptive immunity. Therapeutic agents that indirectly and selectively target Notch signaling in breast cancer cells would be a potentially attractive strategy. However, no such agents have been identified to date. We have found that the MAPK5-ERK5 kinase pathway, which contains at least two druggable targets, functions as a master regulator of Notch signaling in TNBC cells. ERK5 knockout TNBC cells have dramatically decreased expression of Notch receptors, ligands and transcriptional targets. In vivo, these cells form barely detectable tumors that do not metastasize and express lower levels of Notch1 and its ligand Jagged1. Using in silico screening, we identified a class of compounds that selectively target MAP2K5 (MEK5) and decrease the phosphorylation of MAPK7 (ERK5). We selected compound SC-181 for further study. Consistent with ERK5KO cells, pharmacological suppression of ERK5 phosphorylation with SC-181 decreased Notch1 and Jagged1 mRNAs and proteins. SC-181 reversed EMT and reduced the CD44hi/CD24lo CSC population in TNBC cells, but had no effect on T-cell proliferation. SC-181 decreased the number and size of mammospheres in a concentration-dependent manner. Overexpression of the Notch1 intracellular domain (N1IC) in ERK5KO cells rescues their phenotype, dramatically increasing the CSC fraction and promoting EMT. Our results suggest that targeting the MEK5-ERK5 pathway is a promising new strategy to selectively modulate Notch signaling in TNBC CSC without compromising tumor immunity.

Citation Format: Ucar DA, Matossian MD, Hoang-Barnes VT, Hossain FM, Gupta M, Burks HE, Wright TD, Cavanaugh J, Flaherty P, Burow ME, Miele L. A novel druggable target upstream of Notch: MEK5/ERK5 signaling regulates Jagged-1 and Notch1 expression in triple negative breast cancer stem cells [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-03-04.
Abstract P2-03-08: Preclinical evaluation of the PI3Kα/δ inhibitor, copanlisib in HER2+ breast cancer: A proof of concept study
Authors: De P , Carlson JH , Dey N , Leyland-Jones B
Published: May 2019

Purpose: The PI3K-AKT-mTORC1/C2 pathway is frequently activated in HER2+ breast cancer and upregulation of this pathway is a key mechanism of trastuzumab resistance. However, attempts to indirectly target this pathway by using the allosteric mTOR inhibitor everolimus have had limited clinical success. Here, we present the results of a preclinical study of the PI3K α/δ (dominant) inhibitor copanlisib alone and in combination with T-DM1, in HER2 amplified cell lines including a model with acquired resistance to trastuzumab.

Method: Anti-proliferative, apoptotic, cell cycle, and intracellular signaling effects of copanlisib alone and in combination with T-DM1 were evaluated in a panel of HER2 amplified (ER+ or ER-), and HER2 amplified/PIK3CA mutated cell lines, as well as trastuzumab-resistant breast cancer cell lines.

Results: 1) Copanlisib inhibited PI3K, mTOR and their downstream signaling molecules in HER2 amplified/PIK3CA WT or PIK3CA mutated as well as in trastuzumab-resistant cell lines, 2) interestingly, copanlisib also inhibited RAS-MAPK signaling in the earlier time points, 3) copanlisib caused a strong differential growth inhibition in HER2 amplified BC cell lines by 3D-ON-TOP clonogenic assay and real-time monitoring in an IncuCyte Zoom. Inhibition was greater when copanlisib was combined with T-DM1, 4) administration of copanlisib induced cell cycle G0/G1 arrest and resulted in increased apoptosis in a dose-dependent manner, 5) unlike everolimus copanlisib blocked HIF1α accumulation in hypoxic condition and this blocking effect was reversed by prior treatment with the proteasome inhibitor, carfilzomib and 6) copanlisib also attenuated HER2 amplified cell migration, an important phenotypic feature for metastasis.

Conclusions: Copanlisib is highly effective (blocks proliferation, induces apoptosis, and inhibits PI3K and its downstream signaling targets) in HER2 amplified breast cancer cell lines including trastuzumab-resistant and PIK3CA mutated cell lines. The addition of copanlisib to T-DM1 might represent an improved treatment strategy for patients with refractory metastatic HER2+ breast cancer.

Citation Format: De P, Carlson JH, Dey N, Leyland-Jones B. Preclinical evaluation of the PI3Kα/δ inhibitor, copanlisib in HER2+ breast cancer: A proof of concept study [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-03-08.
Abstract P2-03-10: Not presented
Authors: Mallinger A , Dalenc F , Poirot M , Silvente-Poirot S
Published: May 2019

This abstract was not presented at the conference.

Citation Format: Mallinger A, Dalenc F, Poirot M, Silvente-Poirot S. Not presented [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-03-10.
Abstract P2-05-04: The WNT pathway gene, LBH, is critically required for breast carcinogenesis by promoting tumor initiation and survival
Authors: Garikapati K , Chen P , Ashad-Bishop K , Briegel K
Published: May 2019

Background: Cancer stem cells (CSCs) are a small subset of tumor cells with stem cell properties that drive tumor initiation, treatment resistance, and metastasis, thereby leading to rapid disease relapse. However, no cancer stem cell specific treatments are currently available in the clinic. Thus, there is a need to identify novel cancer stem cell-specific molecular targets.

LBH (Limb-Bud and Heart), is a poorly characterized transcription co-factor in the WNT pathway, a signaling network that is key to normal stem cell control and cancer stem cell transformation. We previously identified LBH as a basal mammary stem cell specific marker critically required for stem cell self-renewal and maintenance during normal mammary gland development. Significantly, LBH is also abnormally over-expressed specifically in worst prognosis basal-like breast cancers, a treatment-resistant breast cancer subtype that is enriched in cancer stem cells. Moreover, high inter-tumor LBH expression levels correlate with reduced metastasis-free patient survival and increased chemo-resistance. However, the role of LBH in breast carcinogenesis is not known.

Results: To identify the potential role of LBH in breast carcinogenesis, we abrogated LBH expression in breast cancer cell lines with high CSC populations (HCC1395, MDA-MB-231) using shRNA and CRISPR/Cas technologies. Conversely, we stably introduced LBH into more differentiated breast cancer cell lines with low CSC populations (MCF7, BT549). We found that LBH depletion significantly reduced, while LBH overexpression increased in vitro tumor sphere formation and clonogenic ability, suggesting LBH promotes a CSC phenotype. Modulation of LBH expression did not change proliferation rates, indicating the CSC promoting effects of LBH were not due to increased cell proliferation. However, LBH reduced apoptosis rates and increased tumor cell survival. Importantly, loss of LBH markedly decreased in vivo tumor initiation and metastatic potential of triple negative breast cancer cells, which are important functions of CSC. The underlying mechanisms will be discussed.

Conclusions: Collectively our results suggest that LBH is required for breast CSC self-renewal and maintenance. Thus, LBH inhibition may have value for future CSC-targeted anti-cancer therapy.

Citation Format: Garikapati K, Chen P, Ashad-Bishop K, Briegel K. The WNT pathway gene, LBH, is critically required for breast carcinogenesis by promoting tumor initiation and survival [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-05-04.
Abstract P2-05-08: An assessment of the potential role of intracellular Ca2+ store regulators in breast cancer cells
Authors: Bong AH , Roberts-Thomson SJ , Milevskiy MJ , Monteith GR
Published: May 2019

Background: Calcium (Ca2+) regulates many crucial cellular processes including cell survival, proliferation and death. Endoplasmic reticulum (ER) Ca2+ store levels are critical in the death-inducing effects of some anti-cancer agents. Modulators of ER Ca2+ signalling, such as neuronal calcium sensor-1 (NCS-1) may therefore represent new therapeutic opportunities to enhance the effect of some anti-cancer agents. NCS-1 is associated with poorer survival in breast cancer patients. However, the expression of NCS-1 in specific breast cancer molecular subtypes and its potential role in intracellular Ca2+ signalling in breast cancer cells has not been fully explored.

Aim: To assess expression of NCS-1 in breast cancer molecular subtypes and assess the effect of silencing NCS-1 on intracellular Ca2+ homeostasis and on sensitivity to doxorubicin treatment in MDA-MB-231 breast cancer cells.

Methods: NCS-1 levels were assessed in breast cancer molecular subtypes based on PAM50 subtyping using the TCGA public breast cancer database. Intracellular Ca2+ changes as a consequence of siRNA-mediated silencing of NCS-1 were evaluated using a Fluorescence Imaging Plate Reader (FLIPR) in MDA-MB-231 cells expressing the genetically-encoded Ca2+ indicator GCaMP6m. The effect of NCS-1 silencing on MDA-MB-231 cells treated with doxorubicin (0.03 and 1 μM, 24 h) was evaluated by cell nuclear enumeration (Hoechst 33342 staining) and the percentage of dead cells (propidium iodide staining). Images were acquired using an automated epifluorescence microscope (ImageXpress Micro).

Results: Levels of NCS-1 were higher in the basal molecular subtype compared to other molecular subtypes. NCS-1 silencing promoted cell death induced by 1 μM doxorubicin. NCS-1 silencing had no major effect on cytosolic free Ca2+ levels as a result of either IP3-mediated Ca2+ store release with the purinergic receptor activator ATP or the protease activated receptor activator trypsin. However, NCS-1 silencing suppressed constitutive Ca2+ influx in MDA-MB-231 breast cancer cells. The expression of NCS-1 was positively correlated with the Ca2+ entry channel Orai1 in breast cancers on the TCGA database. Orai1 is associated with increased migration and invasiveness in some breast cancers.

Discussion: These studies suggest that elevation in NCS-1 is a feature of breast cancers of the basal molecular subtype and that NCS-1 is a regulator of the cytotoxic effects of doxorubicin. Further studies are required to determine if this effect is related to changes in constitutive Ca2+ influx that may be mediated via Orai1.

Citation Format: Bong AH, Roberts-Thomson SJ, Milevskiy MJ, Monteith GR. An assessment of the potential role of intracellular Ca2+ store regulators in breast cancer cells [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-05-08.
Abstract P2-05-09: Withdrawn
Authors: Connelly L , Tsang Mui Chung S
Published: May 2019

This abstract was withdrawn by the authors.

Citation Format: Connelly L, Tsang Mui Chung S. Withdrawn [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-05-09.
Abstract P2-06-10: Withdrawn
Authors: Sood AK , Mukhopadhyay A , Wang J
Published: May 2019

This abstract was withdrawn by the authors.

Citation Format: Sood AK, Mukhopadhyay A, Wang J. Withdrawn [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-06-10.
Abstract P2-06-13: A novel small molecule JMX0293 inhibits the growth of triple-negative breast cancer via suppressing STAT3 and inducing apoptosis
Authors: Dong J , Xu J , Kim H , Zhou J , Shen Q
Published: May 2019

Approximately 30-40% of breast cancer cases are estrogen receptor negative (ENBC), including the most aggressive triple-negative subtype (TNBC) which spread early and recur frequently with a poor prognosis. ENBCs do not respond to selective ER modulators such as tamoxifen or aromatase inhibitors, and therefore, there is an urgent need of non-ER-based therapies for ENBC and TNBC. We recently developed a novel small molecule JMX0293 based on our lead drug candidate HJC0152. Both HJC0152 and JMX0293 were able to suppress cell proliferation of both ER+ (MCF-7, T-47D) and ER- (MDA-MB-231 and MDA-MB-468) breast cancer cell lines in a dose-dependent manner, with significantly lower toxicity against normal mammary epithelial cells (MCF-10A) in vitro. JMX0293 exhibited moderately improved anti-proliferative effects against highly invasive TNBC cell line MDA-MB-231 and ER+ breast cancer cell line MCF-7 with IC50 values of 0.92 and 2.24 μM, respectively. JMX0293 was also demonstrated to induce apoptosis of MDA-MB-231 cells dose-dependently. Mechanistically, JMX0293 inhibited STAT3 phosphorylation at Tyr705 residue, leading to the down-regulation of Bcl-2 and upregulation of apoptotic proteins including Bax, cleaved-caspase 3 and cleaved-PARP. In addition, JMX0293 reduced the ratio of Bcl-2 to Bax. In vivo, JMX0293 treatment for 2 weeks at a dose of 10 mg/kg significantly suppressed the growth of TNBC xenograft tumors in mice with minimal impact on general health and body weight. Our results suggest that JMX0293 induces TNBC cell apoptosis and contributes to its anti-proliferative effect through inhibiting transcription factor STAT3, and JMX0293 could be further characterized and optimized for developing a promising therapeutic for treating ENBC and TNBC. This work was supported by grant (R01 CA231150) from the NIH/NCI.

Citation Format: Dong J, Xu J, Kim H, Zhou J, Shen Q. A novel small molecule JMX0293 inhibits the growth of triple-negative breast cancer via suppressing STAT3 and inducing apoptosis [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-06-13.
Abstract P2-06-17: A novel interaction of AURKA with MAPK pathway in breast cancer cells as a potential therapeutic target
Authors: Gandhi S , Gil M , Khoury T , Takabe K , Puzanov I , Gelman I , D'Assoro A , Opyrchal M
Published: May 2019

Background: Aurora A (AURKA) is a mitotic kinase responsible for centrosome segregation and mitotic spindle formation. In normal cells, expression of AURKA is highly regulated and is predominantly restricted to G2/M phases of the cell cycle. Unlike healthy cells, cancer cells overexpress AURKA through all phases of the cell cycle resulting in the acquisition of alternate non-mitotic functions. Little is known about cellular functions regulated by AURKA and its interaction with other signaling molecules. Here, we report a novel interaction between AURKA and the mitogen-activated protein kinase (MAPK) pathway at the level of MEK1 in breast cancer cells. This interaction may serve as a novel target as well as demonstrate by an additive cytotoxic effect of AURKA- and MEK1/2-specific inhibitors against estrogen positive (ER+) and triple negative breast cancer (TNBC) cells.

Results: We show that treatment of ER+ HER2- MCF-7, ER- HER2+ SKBR3 and ER- HER2- BT549 cells with AURKA specific inhibitors alisertib, MK8745 and Aurora A Inhibitor I resulted in over 2-fold increase in relative levels of poMEK1/2 and poERK1/2 compared to untreated controls. The activation of the MAPK pathway was rapid with changes seen within 5 min after treatment with AURKA inhibitors and was sustained for at least 48 hours. Treatment with the pan RAF inhibitor TAK-632 did not diminish alisertib-induced poERK and poMEK1/2. Alternatively, treatment with the MEK1/2 specific inhibitor PD0325901 completely abrogated alisertib-induced phosphorylation of MEK1/2 and ERK1/2. In situ proximity ligation and pull down assays demonstrated AURKA and MEK1/2 direct interaction. In vitro kinase assay showed direct phosphorylation of MEK1 by AURKA. Combined treatment of alisertib and PD0325901 in vitro revealed significant additive cytotoxic effect in MCF-7 and BT549 cells when compared to either agent used alone (p< 0.008 and p<0.011; p <0.04 and p<0.028) with early trend toward significance in survival in a BT549 xenograft breast cancer in vivo model.

Conclusions: Our data shows a novel AURKA-MEK1 interaction in breast cancer cells. In depth in vivo analysis is ongoing. The results reveal a promising new strategy for the treatment TNBC patients using a combination of AURKA and MEK1/2 inhibitors.

Citation Format: Gandhi S, Gil M, Khoury T, Takabe K, Puzanov I, Gelman I, D'Assoro A, Opyrchal M. A novel interaction of AURKA with MAPK pathway in breast cancer cells as a potential therapeutic target [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-06-17.
Abstract P2-06-18: Withdrawn
Authors: Kunder R , Fernandez Dunne S , Clutter M , Cristofanilli M , Mendillo M , Vasilopoulos A , Horiuchi D
Published: May 2019

This abstract was withdrawn by the authors.

Citation Format: Kunder R, Fernandez Dunne S, Clutter M, Cristofanilli M, Mendillo M, Vasilopoulos A, Horiuchi D. Withdrawn [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-06-18.
Abstract P2-06-20: CDC20 is a novel therapeutic target in triple negative breast cancer
Authors: Sharma J , Raicu I , Nguyen J , Udden N , Wang Q , Alluri P
Published: May 2019

Background: Approximately 15% of breast cancers lack expression of Estrogen Receptor (ER), Progesterone Receptor (PR) and Human Epidermal Growth Factor Receptor 2 (HER-2), and are referred to as Triple Negative Breast Cancer (TNBC). Clinical outcomes in patients with TNBC continue to be poor due to inherently aggressive biology of the disease and paucity of targeted therapies. In this study, we identified Cell Division Cycle protein 20 (CDC20), a key regulatory protein that mediates sister chromatid separation in mitosis, as a novel therapeutic target in TNBC.

Methods: To uncover new drivers of TNBC, we nominated genes that are significantly overexpressed in patients with TNBC, compared to non-TNBCs. After identifying CDC20 as a top hit in this analysis, we used expression microarrays, RNA-seq analyses and Western blotting to assess CDC20 expression levels in breast cancer patients and cell lines. Kaplan-Meier analyses were performed to study the correlation between CDC20 expression levels and clinical outcomes, including recurrence-free survival, metastasis-free survival and overall survival. shRNA-mediated knockdown of CDC20 in TNBC cell lines was employed to study the role of CDC20 in TNBC proliferation, invasion, migration and xenograft growth in mice.

Results: CDC20 was significantly overexpressed in TNBCs (compared to non-TNBCs) in The Cancer Genome Atlas (TCGA) breast dataset, and in several other independent breast cancer datasets, including Curtis, Stickeler and Kao. CDC20 expression was also significantly higher in TNBC cell lines compared to non-TNBC cell lines. Furthermore, expression levels of CDC20 were highly significantly correlated with clinical outcomes, including recurrence-free survival, metastasis-free survival and overall survival. shRNA-mediated knockdown of CDC20 expression in TNBC cell lines, MDA-MB-231 and MDA-MB-468, resulted in decreased proliferation, invasion and migration. Our preliminary findings suggest that CDC20 knockdown also results in inhibition of cell line-derived xenograft growth in mice.

Conclusion: We identified CDC20 is a novel therapeutic target in TNBC. Our findings support development of pharmacologic strategies to inhibit CDC20 function as a potential targeted therapy against TNBC.

Citation Format: Sharma J, Raicu I, Nguyen J, Udden N, Wang Q, Alluri P. CDC20 is a novel therapeutic target in triple negative breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-06-20.
Abstract P2-07-11: Prediction of the Oncotype Dx recurrence score (RS) from clinicopathologic factors
Authors: Voutsadakis IA , Thibodeau S , Reed M
Published: May 2019

Background: The Oncotype Dx assay is currently used as an aid to therapeutic decisions for the adjuvant treatment of women with ER-positive, Her2-negative, lymph node-negative or -micrometastatic breast cancer. The recently reported TAILORx study showed that no or minimal benefit is derived from adjuvant chemotherapy in patients with an Oncotype Dx Recurrence Score (RS) of 25 or less.

Methods: Charts of breast cancer patients that had the Oncotype Dx test in our cancer center in a nine-year period were reviewed. Data on demographic, and cancer-specific characteristics of the included patients were extracted. Predicted disease recurrence from the Oncotype Dx test was recorded and correlated with select clinicopathologic characteristics.

Results: Two hundred and thirty patients with ER-positive, Her2-negative, lymph node-negative or micrometastatic breast cancer were included. Mean age was 65 years-old (SD 9.9). Two hundred and three patients (88.3%) were post-menopausal and one hundred and thirty-three patients (57.8%) were 65 years-old or older. Two hundred and nine patients (90.9%) had lymph node-negative disease. Oncotype Dx recurrence score was low (<11) in sixty-four patients (27.8%), intermediate (11-25) in one hundred and forty patients (60.9%) and high (>25) in twenty-six patients (11.3%). High tumor grade and low progesterone receptor (PR) staining by IHC were the two clinicopathologic factors most associated with a high Oncotype Dx RS (x2 test p <0.00001 and Fisher's exact test p <0.0001). A predictive index (PI) was constructed, assigning one point each for grade 3 and PR staining in 20% or less of tumor cells. A PI of 0 was observed in one hundred and thirty-eight patients (60%), a PI of 1 was observed in seventy-one patients (30.9%), and a PI of 2 was observed in twenty-one patients (9.1%). One hundred and thirty-four patients (97.1%) with a PI of 0 had a RS of 25 or less. Patients with a PI of 1 and 2 had a RS of >25 in 12.7% and 61.9% of cases, respectively.

Conclusion: The PI based on tumor grade and PR we propose is a simple predictor of Oncotype Dx RS. 97.1% of patients with a grade 1 or 2 tumor and PR positivity in >20% of tumor cells had a RS of 25 or less. The Oncotype Dx test and its associated cost can therefore be avoided in these patients, especially in low-resource settings.

Citation Format: Voutsadakis IA, Thibodeau S, Reed M. Prediction of the Oncotype Dx recurrence score (RS) from clinicopathologic factors [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-07-11.
Abstract P2-08-05: Withdrawn
Authors: Jaraj D , Ahlgren J , Arnesson L-G , Einbeigi Z , Höijer J , Klintman M , Malmström P , Vikhe Patil E , Sund M , Fredriksson I , Bergh J , Pettersson A
Published: May 2019

This abstract was withdrawn by the authors.

Citation Format: Jaraj D, Ahlgren J, Arnesson L-G, Einbeigi Z, Höijer J, Klintman M, Malmström P, Vikhe Patil E, Sund M, Fredriksson I, Bergh J, Pettersson A. Withdrawn [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-08-05.
Abstract P2-08-27: Not presented
Authors: Rayen JM , Poole GH , Rhodes C
Published: May 2019

This abstract was not presented at the conference.

Citation Format: Rayen JM, Poole GH, Rhodes C. Not presented [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-08-27.
Abstract P2-08-28: Relationship between FDG uptake and neutrophil/lymphocyte ratio in patients with breast invasive ductal cancer
Authors: Fujii T , Yanai K , Tokuda S , Nakazawa Y , Kurozumi S , Obayashi S , Yajima R , Hirakata T
Published: May 2019

Background: 18F-Fluorodeoxyglucose-positron emission tomography (FDG-PET) is used to evaluate the glucose metabolic rates of cancers. Several studies have reported that high FDG uptake is predictive of poor prognosis and aggressive features in patients with breast cancer. FDG uptake is influenced by many factors, including inflammation. In this study, we investigated the relationship between FDG uptake and neutrophil/lymphocyte ratio (NLR), which is an indicator of systemic inflammation. Patients and Methods: We retrospectively investigated the cases of 143 consecutive invasive ductal carcinoma patients who had undergone surgery and FDG-PET preoperatively.PET was evaluated using standardized uptake value max (SUVmax). The median SUVmax was 2.5 (range 0-10.5). Thus, we divided the cases into two groups based on the value of SUVmax; low (<2.5) and high (≥2.5). The relationships between SUVmax and clinicopathological features, including NLR, were investigated. Results: Among the143 patients, 73 (51.0%) had high SUVmax in the primary tumor. The analysis revealed that large tumor size (p<0.001), high nuclear grade (p<0.001), the presence of lymphovascular invasion (p<0.001), CRP (p=0.046) and high NLR (p<0.001) were significantly associated with high SUVmax in the primary tumor. There were associations between SUVmax and NLR (r=0.323, p<0.001). Among the 70 cases with low SUVmax, there was no recurrent disease, while 6 cases among the 73 cases with high SUVmax had disease recurrence. It is interesting to note that the group with high SUVmax and low NLR had no recurrent disease. Conclusions: To the best of our knowledge, this is the first report to describe the relationship between FDG uptake and NLR in breast cancer. The present study demonstrated that the finding of preoperative high FDG uptake in breast cancer may be reflective of poor prognosis and that high NLR may be predictive of aggressive features among patients with breast cancer. On the other hand, among breast cancer patients with high SUVmax in the primary tumor, it will be useful to identify the subset of patients with low NLR in order to improve prognostic accuracy.

Citation Format: Fujii T, Yanai K, Tokuda S, Nakazawa Y, Kurozumi S, Obayashi S, Yajima R, Hirakata T. Relationship between FDG uptake and neutrophil/lymphocyte ratio in patients with breast invasive ductal cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-08-28.
Abstract P2-08-29: The impact of time interval between diagnosis and surgery in each type and stage of breast cancer
Authors: Kim J-M , Choi HJ , Kim I , Ryu JM , Yu J , Lee JE , Kim SW , Nam SJ , Lee SK
Published: May 2019

Background: There are many factors that might contribute to the delay of surgery in patients with breast cancer. Previous studies investigate the influence of delay of surgery, but they reported inconsistent results. The purpose of this study was to evaluate the impact of time of surgery on prognosis of breast cancer.

Methods: We performed a retrospective review of the patients with breast cancer, who received surgery between 1992 and 2009, by using data from Korea Breast Cancer Society Registry. Kaplan-Meier survival analysis and Cox regression model were used to evaluate the impact of time to surgery in breast cancer and subgroup analyses were performed for each disease stage and molecular subtype.

Result: A total 14727 patients were included for analysis. Delay of surgery more than 31 days was associated with worse survival for breast cancer [hazard ratio (HR) = 2.16; 95% confidence interval (CI), 1.936-2.408, p<0.001]. Subgroup analyses revealed that over 31 days of surgical delay were significantly associated with worse survival in hormone receptor positive and HER-2 negative (p<0.001), hormone receptor positive and HER-2 positive (p<0.001), hormone receptor negative and HER-2 positive (p<0.001), triple negative (p<0.001) and stage II, III breast cancer patients (p<0.001).

Conclusion: Surgical delay of more than 31 days were independent risk factors for worse outcome of breast cancer in each molecular subtype and breast cancer group except stage 0 and I. Although preoperative evaluation is required, surgical delay should be shortened to enhance survival of breast cancer, especially in patients with tumor size more than 2cm or presence of lymph node metastasis.

Citation Format: Kim J-M, Choi HJ, Kim I, Ryu JM, Yu J, Lee JE, Kim SW, Nam SJ, Lee SK. The impact of time interval between diagnosis and surgery in each type and stage of breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-08-29.
Abstract P2-08-35: A significance of SUVmax levels on FDG-PET as a prognostic factor may be mediated by local immune environment of breast cancer
Authors: Fujimoto Y , Higuchi T , Watanabe T , Hida AI , Imamura M , Kitajima K , Miyoshi Y
Published: May 2019

(Background) SUVmax levels (maximum radioactivity concentration per a pixel) on FDG-PET reflect glucose uptake and it is clinically useful as a prognostic factor. It is reported that breast cancer with high levels of SUVmax causes insufficient glucose concentration in stromal tissue, which results in suppressed cytotoxic T-lymphocytes function. These data may indicate that the prognostic significance of SUVmax levels is influenced by local immune environment of breast cancer. The aim of this study is to investigate whether local immune responses of breast cancer affect correlation of SUVmax levels and prognosis.

(Method) The 278 invasive breast cancer patients were recruited who underwent surgery at Hyogo College of Medicine Hospital and whose SUVmax levels in the breast were examined before surgery or neo-adjuvant therapy. The cutoff value of SUVmax levels was set at 3.585. Tumor infiltrate lymphocytes (TILs) were evaluated as a local immune response and the distributions of TILs were divided into three groups, inflamed (intra-tumoral lymphocytes, Inf), immune excluded (peri-tumoral lymphocytes, IE) and immune desert (very few lymphocytes, ID). During follow-up period (median 39 months), 21 patients relapsed.

(Results) Relapse free survival (RFS) in the SUVmax-high group was significantly worse than in the SUVmax–low group (p=0.0026). There was no correlation between TILs distribution patterns and RFS. In the IE+ID group (175 patients) SUVmax levels were not correlated with prognosis, but in the Inf group (103 patients) RFS of SUVmax-high was significantly worse than of SUVmax-low (p=0.0051). In the multivariate analysis including nodal status and nuclear grade, SUVmax levels of the Inf group was an independent prognostic factor.

(Discussion and conclusion) SUVmax levels in primary lesions were correlated with prognosis only in the Inf group and were not in the IE+ID group. A significance of SUVmax levels as a prognostic factor may be diverse depending on the local immune environment of breast cancer. A novel therapeutic strategy such as inducing suppression of glucose uptake in cancer cells is suggested for breast cancer with immune inflamed.

Citation Format: Fujimoto Y, Higuchi T, Watanabe T, Hida AI, Imamura M, Kitajima K, Miyoshi Y. A significance of SUVmax levels on FDG-PET as a prognostic factor may be mediated by local immune environment of breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-08-35.
Abstract P2-08-37: The prognostic impact of synaptojanin 2 expression in estrogen receptor α-positive breast cancer patients
Authors: Nishikawa S , Kondo N , Wanifuchi-Endo Y , Hisada T , Uemoto Y , Katagiri Y , Dong Y , Kato H , Takahashi S , Toyama T
Published: May 2019

Background: Synaptojanin 2 (SYNJ2) was reported to be a druggable mediator of metastasis. It is overexpressed and amplified in breast cancer, particularly estrogen receptor α (ERα)-positive breast cancer. SYNJ2 was also shown to promote cell migration and invasion in breast cancer xenograft cultures and lung metastasis in mice. Here, we investigated SYNJ2 mRNA expression in breast cancer patients during long-term follow-up.

Materials and methods: A total of 434 invasive breast cancer tissues were analyzed for SYNJ2mRNA expression using TaqMan PCR, and the correlation of this expression with patient clinicopathological factors was determined. We also examined the expression of markers associated with tumor-initiating capacity, such as SNAI1, SNAI2, and VIM. Survival curves were analyzed using the Kaplan–Meier method. Cox proportional hazards regression analysis was used for univariate and multivariate analyses of prognostic values.

Results: The median follow-up period was 10.7 years. We found positive correlations between high expression of SYNJ2 mRNA and shorter disease-free survival in breast cancer patients (P=0.049), which was limited to ERα-positive patients (P=0.020) and not seen in ERα-negative patients (P=0.863). High SYNJ2 mRNA levels were positively correlated with high tumor grade, ERα negativity, and node positivity. Multivariate analysis indicated that high SYNJ2 mRNA expression was an independent factor for poor disease-free survival in breast cancer patients.

Multivariate analysis of poor disease-free survival   DFS    Mutivariate   All patientsp valueHR (95%CI)Tumor size2cm and fewer1540.43151 (Reference) more than 2cm280 1.19 (0.78-1.85)Lymph node metastasisNegative2190.0001 and fewer1 (Reference) Positive184 5.61 (3.59-9.05) Unknown31 2.40 (0.89-5.48)Grade1,22330.38471 (Reference) 3195 1.20 (0.79-1.81) Unknown6 0.17 (1.78-3.49)ER statusPositive3370.02621 (Reference) Negative97 2.06 (1.87-4.07)PgR statusPositive2930.39511 (Reference) Negative141 1.30 (0.70-2.28)SYNJ2 mRNA expressionlow2170.02251 (Reference) high217 1.57 (1.07-2.34)

Conclusion:High SYNJ2 expression was shown to be an independent predictive factor of poor prognosis in ERα-positive breast cancer patients. SYNJ2 could therefore be used as a candidate biomarker and therapeutic target in breast cancer.

Citation Format: Nishikawa S, Kondo N, Wanifuchi-Endo Y, Hisada T, Uemoto Y, Katagiri Y, Dong Y, Kato H, Takahashi S, Toyama T. The prognostic impact of synaptojanin 2 expression in estrogen receptor α-positive breast cancer patients [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-08-37.
Abstract P2-08-44: Not presented
Authors: Allemand C , Calvo MF , Valerio AC , Riggi MC , Corrao F , Orti R , Wernicke A , Zamora L , Paez MC , Lorusso C
Published: May 2019

This abstract was not presented at the conference.

Citation Format: Allemand C, Calvo MF, Valerio AC, Riggi MC, Corrao F, Orti R, Wernicke A, Zamora L, Paez MC, Lorusso C. Not presented [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-08-44.