Search

Purpose: Cyclin-dependent kinase 4 (CDK4) and CDK6 inhibitors (CDK4/6i) are highly effective against estrogen receptor–positive (ER+)/HER2− breast cancer; however, intrinsic and acquired resistance is common. Elucidating the molecular features of sensitivity and resistance to CDK4/6i may lead to identification of predictive biomarkers and novel therapeutic targets, paving the way toward improving patient outcomes. Experimental Design: Parental breast cancer cells and their endocrine-resistant derivatives (EndoR) were used. Derivatives with acquired resistance to palbociclib (PalboR) were generated from parental and estrogen deprivation–resistant MCF7 and T47D cells. Transcriptomic and proteomic analyses were performed in palbociclib-sensitive and PalboR lines. Gene expression data from CDK4/6i neoadjuvant trials and publicly available datasets were interrogated for correlations of gene signatures and patient outcomes. Results: Parental and EndoR breast cancer lines showed varying degrees of sensitivity to palbociclib. Transcriptomic analysis of these cell lines identified an association between high IFN signaling and reduced CDK4/6i sensitivity; thus an “IFN-related palbociclib-resistance Signature” (IRPS) was derived. In two neoadjuvant trials of CDK4/6i plus endocrine therapy, IRPS and other IFN-related signatures were highly enriched in patients with tumors exhibiting intrinsic resistance to CDK4/6i. PalboR derivatives displayed dramatic activation of IFN/STAT1 signaling compared with their short-term treated or untreated counterparts. In primary ER+/HER2− tumors, the IRPS score was significantly higher in lumB than lumA subtype and correlated with increased gene expression of immune checkpoints, endocrine resistance, and poor prognosis. Conclusions: Aberrant IFN signaling is associated with intrinsic resistance to CDK4/6i. Experimentally, acquired resistance to palbociclib is associated with activation of the IFN pathway, warranting additional studies to clarify its involvement in resistance to CDK4/6i.

Cancer immunology is the most rapidly expanding field in cancer research, with the importance of immunity in cancer pathogenesis now well accepted including in the endocrine-related cancers. The immune system plays an essential role in the development of ductal and luminal epithelial differentiation in the mammary gland. Originally identified as evolutionarily conserved antipathogen cytokines, interferons (IFNs) have shown important immune-modulatory and antineoplastic properties when administered to patients with various types of cancer, including breast cancer. Recent studies have drawn attention to the role of tumor- and stromal-infiltrating lymphocytes in dictating therapy response and outcome of breast cancer patients, which, however, is highly dependent on the breast cancer subtype. The emerging role of tumor cell–inherent IFN signaling in the subtype-defined tumor microenvironment could influence therapy response with protumor activities in breast cancer. Here we review evidence with new insights into tumor cell–intrinsic and tumor microenvironment–derived IFN signaling, and the crosstalk of IFN signaling with key signaling pathways in estrogen receptor–positive (ER+) breast cancer. We also discuss clinical implications and opportunities exploiting IFN signaling to treat advanced ER+ breast cancer.

Despite effective strategies, resistance in HER2+ breast cancer remains a challenge. While the mevalonate pathway (MVA) is suggested to promote cell growth and survival, including in HER2+ models, its potential role in resistance to HER2-targeted therapy is unknown. Parental HER2+ breast cancer cells and their lapatinib-resistant and lapatinib + trastuzumab–resistant derivatives were used for this study. MVA activity was found to be increased in lapatinib-resistant and lapatinib + trastuzumab–resistant cells. Specific blockade of this pathway with lipophilic but not hydrophilic statins and with the N-bisphosphonate zoledronic acid led to apoptosis and substantial growth inhibition of R cells. Inhibition was rescued by mevalonate or the intermediate metabolites farnesyl pyrophosphate or geranylgeranyl pyrophosphate, but not cholesterol. Activated Yes-associated protein (YAP)/transcriptional coactivator with PDZ-binding motif (TAZ) and mTORC1 signaling, and their downstream target gene product Survivin, were inhibited by MVA blockade, especially in the lapatinib-resistant/lapatinib + trastuzumab–resistant models. Overexpression of constitutively active YAP rescued Survivin and phosphorylated-S6 levels, despite blockade of the MVA. These results suggest that the MVA provides alternative signaling leading to cell survival and resistance by activating YAP/TAZ–mTORC1–Survivin signaling when HER2 is blocked, suggesting novel therapeutic targets. MVA inhibitors including lipophilic statins and N-bisphosphonates may circumvent resistance to anti-HER2 therapy warranting further clinical investigation. Implications: The MVA was found to constitute an escape mechanism of survival and growth in HER2+ breast cancer models resistant to anti-HER2 therapies. MVA inhibitors such as simvastatin and zoledronic acid are potential therapeutic agents to resensitize the tumors that depend on the MVA to progress on anti-HER2 therapies.

BACKGROUND The os trigonum is an accessory bone that is not fully fused with the talus during secondary ossification, and is one of the risk factors of posterior malleolus impact syndrome. The purpose of this study was to classify the os trigonum and to guide the diagnosis and treatment of related clinical diseases. MATERIAL AND METHODS Ankle computed tomography (CT) scans of 586 Chinese patients between October 2014 and October 2018 were reviewed. CT images of 1011 ankle joints were reconstructed to evaluate the classification of the os trigonum and the measurement of anatomical parameters. RESULTS The incidences of os trigonum in 3 groups were determined as type I (1.9%), type II (10.5%), and type III (14.7%). The macro-axis of type II (0.89±0.31) cm was significantly larger than with type I (0.65±0.24 cm) and type III (0.74±0.23 cm) (p

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Endocrine therapy resistance in breast cancer is a major obstacle in the treatment of patients with estrogen receptor-positive (ER+) tumors. Herein, we demonstrate the feasibility of longitudinal, noninvasive and semiquantitative in vivo molecular imaging of resistance to three endocrine therapies by using an inducible fluorescence-labeled short hairpin RNA (shRNA) system in orthotopic mice xenograft tumors. We employed a dual fluorescent doxycycline (Dox)-regulated lentiviral inducer system to transfect ER+ MCF7L breast cancer cells, with green fluorescent protein (GFP) expression as a marker of transfection and red fluorescent protein (RFP) expression as a surrogate marker of Dox-induced tumor suppressor phosphatase and tensin homolog deleted on chromosome 10 (PTEN) knockdown. Xenografted MCF7L tumor-bearing nude mice were randomized to therapies comprising estrogen deprivation, tamoxifen or an ER degrader (fulvestrant) and an estrogen-treated control group. Longitudinal imaging was performed by a home-built multispectral imaging system based on a cooled image intensified charge coupled device camera. The GFP signal, which corresponds to number of viable tumor cells, exhibited excellent correlation to caliper-measured tumor size (P <

Background: Forkhead box A1 (FOXA1) is an essential pioneer transcription factor (TF) evoking other key TFs-mediated lineage-specific transcriptional programs in several endoderm-derived organs. Aberrant FOXA1 augmentation, via genetic alterations, occurs in 10-15% of ER+ primary and metastatic breast cancer (BC). We have recently shown that high levels of FOXA1 (H-FOXA1) induces enhancer and transcriptional reprogramming to promote endocrine-resistant (EndoR) and pro-metastatic phenotypes. Using the core transcriptional regulatory circuitry (CRC) mapping method, we identified the AP-1 TF JUNB as a key CRC component in BC cells expressing H-FOXA1. In this study, we aimed to further characterize key AP-1 components that play a role in mediating H-FOXA1-induced transcriptional reprogramming in EndoR and metastatic BC. Methods: The ROSE and HOMER tools were used to identify super-enhancers (SEs) and the predicted SE-harboring TFs in MCF7-parental (P) cells with ectopic FOXA1 overexpression (OE), and in MCF7 tamoxifen-resistant (TamR) cells with endogenous FOXA1 amplification and OE. Cell growth, migration, and co-immunoprecipitation assays, were performed using ER+ BC P cells with ectopic FRA1 OE. A FOXA1 core gene signature (GS) was deduced using the BETA.plus algorithm to analyze our previously reported RNA-seq and ChIP-seq data derived from MCF7-P cells with ectopic FOXA1 OE. Additional RNA-seq analyses include MCF7-P cells with ectopic FRA1 OE, FOXA1 OE and simultaneous FRA1 siRNA knockdown (KD), and MCF7-TamR cells with FRA1 KD. We identified a FOXA1/FRA1-centered GS and its clinical relevance was examined using expression profiles of TCGA, METABRIC, and a metastatic biopsy study from cohort of patients with ER+ metastatic BC from Dana-Farber Cancer Institute.Results: We identified FRA1 as one of the top TFs selectively harboring SEs at their gene loci in MCF7-TamR vs. P cells. Both FRA1 and JUNB expression was elevated in TamR vs. P cells and altered concordantly with FOXA1 in P and TamR cells upon FOXA1 OE or KD, respectively. As we identified JUNB as a CRC component with binding sites enriched at the SEs in BC cells expressing H-FOXA1, we hypothesized that FRA1 and JUNB form a feed-forward transcriptional axis amplifying H-FOXA1-induced enhancer reprogramming. We found that JUNB co-immunoprecipitated with FRA1 in MCF7-TamR and MCF7-P cells with ectopic FRA1 OE, suggesting that FRA1 forms a heterodimer with JUNB to exert AP-1 activity. Ectopic FRA1 OE reduced P cell endocrine sensitivity, increased cell migration, and elicited a transcriptome enriched for the FOXA1-induced core GS. In P cells with ectopic FOXA1 OE, we identified a FRA1-dependent GS (n = 27) that is highly enriched for interferon signaling. This FOXA1/FRA1 GS was highly expressed in luminal B vs. A subtype of primary tumors, further elevated in ER+ metastases, where its expression was positively correlated with FRA1 mRNA levels. Notably, this FOXA1/FRA1 GS was not dependent on FRA1 in P cells without FOXA1 OE, suggesting its relevance in the context of H-FOXA1.Conclusions: Here we show that a FOXA1/FRA1-centered transcriptional axis induces an interferon signaling-enriched GS associated with poor outcome of ER+ BC and metastasis. A FRA1/JUNB AP-1 complex may form a feed-forward transcriptional axis to amplify H-FOXA1 signaling. The FOXA1/FRA1-centered GS could be used to stratify patients with ER+ BC who may need additional targeted therapies. Further studies are warranted to elucidate the interplay between FOXA1 and FRA1/JUNB in regulating interferon signaling, which may guide approaches to improve patient outcomes, possibly with immunotherapy using immune checkpoint inhibitors. Citation Format: Xiaoyong Fu, Resel Pereira, Lanfang Qin, Carmine De Angelis, Sarmistha Nanda, Martin Shea, Agostina Nardone, Rinath Jeselsohn, Ofir Cohen, Nikhil Wagle, Mothaffar Rimawi, C Kent Osborne, Rachel Schiff. A FOXA1/FRA1-centered transcriptional axis regulates interferon signaling in high FOXA1-associated endocrine-resistant and metastatic breast cancer [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PD8-03.

Forkhead box A1 (FOXA1) is a pioneer factor for the nuclear hormone receptors: estrogen receptor and androgen receptor. FOXA1 plays a major role inducing endocrine resistance in breast cancer (BC) and prostate cancer (PC), the two most prevalent cancers in the United States. In this study, we investigated FOXA1 gene alterations across different race and ethnicity using the TCGA PanCan Atlas dataset for BC and PC patients. The BC and PC dataset included 1084 and 494 patient samples, respectively, profiled for copy number alterations (CNA), gene expression, and mutations. In the BC dataset, the samples were from 877 non-Hispanic (81%), 38 Hispanic (3.5%) and 169 patients with no ethnicity data (15.5%). Among them, the majority of patients were White (n=751, 69.3%) followed by Black/African American (AA) (n=182, 16.8%), Asian (n=60, 5.5%), and American Indian or Alaska Native (n=1, 0.09%). Ninety BC patients (8.3%) had no race information. The PC dataset included 152 non-Hispanic (30.8%), 0 Hispanic, and 342 patients with no ethnicity data (69.2%). The samples obtained for the PC study were from White (n=147, 29.8%), Black/AA (n=7, 1.4%), and Asian (n=2, 0.4%) patients. Majority of the PC patients had no racial information (n=338, 68.4%) recorded. In the BC dataset, the incidence of FOXA1 alterations was 16/1070 (1.5%) CNA 24/1082 (2.2%) high mRNA expression, and 31/1066 (2.9%) mutations. Only amplifications were found within the BC patients. In the PC dataset, there were 15/489 (3.1%) CNA, 16/493 (3.2%) high mRNA expression, and 28/494 (5.7%) mutations reported in FOXA1. Deep deletion was found in one of the PC patients while the rest had amplifications. Due to insufficient numbers of Hispanic patients in the datasets, we compared the incidence of various FOXA1 alterations in White vs. Black/AA population using Fisher's exact test. Only FOXA1 mutation rate was significantly higher (p =0.03) in Blacks/AA (2/7, 28.6%) compared to Whites (5/147, 3.4%) in PC, but not in the BC dataset. Comparing the separate results of the FOXA1 CNA and gene overexpression White vs. Black/AA patients were statistically significant in both BC and PC datasets. A majority of FOXA1 mutations were missense mutations with a few frame shifts in BC and PC. The missense mutation reported in both BC and PC datasets were D226G, D226N, and H247Y. Additional studies are necessary to understand the functional significance of these mutations on the development of cancer. Complete and larger datasets that include the race and ethnicity information from diverse group of patients as well as tumor molecular subtyping are also needed for the assessment of the mechanism of health disparity in BC and PC in minority population in the United States. Citation Format: Jennifer Torres, Hariprasad Thangavel, Xiaoyong Fu, Rachel Schiff, Meghana V. Trivedi. FOXA1 genetic alterations in whites versus blacks or African Americans in breast and prostate cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3508.

Radiofrequency ablation (RFA) is an important standard therapy for cardiac arrhythmias, but direct monitoring of tissue treatment is currently lacking. We demonstrate an RFA catheter integrated with polarization sensitive optical coherence tomography (PSOCT) for directly monitoring the RFA process in real time. The integrated RFA/OCT catheter was modified from a standard clinical RFA catheter and includes a miniature forward-viewing cone-scanning OCT probe. The PSOCT system was validated with a quarter-wave plate while the RFA function of the integrated catheter was validated by comparing lesion sizes with those made with an unmodified RFA catheter. Additionally, the integrated catheter guided catheter-tissue apposition and monitored RFA lesion formation in cardiac tissue in real time. The results show that catheter-tissue contact can be characterized by observing the features of the blood and tissue in the acquired OCT images and that RFA lesion formation can be confirmed by monitoring the change in phase retardance in the acquired PSOCT images. This system demonstrates the feasibility of an integrated RFA/OCT catheter to deliver RF energy and image the cardiac wall simultaneously and justifies further research into use of this technology to aid RFA therapy for cardiac arrhythmias.

Estrogen receptor-positive (ER+) breast cancer is the most common subtype of breast cancer. Endocrine therapy targeting the ER pathway is effective, yet endocrine resistance is prevalent and remains a clinical challenge. The mechanisms underlying endocrine resistance are multifaceted and are likely to continue evolving over time in response to various endocrine regimens. The expression of ER in most endocrine-resistant tumors underscores ER’s continuing role, although altered, often via crosstalk with hyperactive growth factor receptor and intracellular kinase signaling pathways. These interactions can alter ER’s sensitivity to various endocrine agents and lead to the activation of distinct transcriptional programs that provide proliferation and survival signaling to escape endocrine therapy. Additional molecular determinants inflicting ER transcriptional reprogramming to promote endocrine resistance include alterations of ER coregulators and pioneer factors, and genetic aberrations of ER itself. Recent advances in our understanding of the mechanisms of endocrine resistance, mostly provided by large-scale sequencing studies, further establish the roles of epigenetic alterations, the DNA damage response, the tumor microenvironment, and the immune response in promoting the endocrine-resistant ER+ disease. Progress has been made in translating several of these findings into effective new targeted therapies, such as inhibitors targeting the key signaling node mTOR and the cyclin-dependent kinases CDK4 and CDK6. However, considerable challenges remain in (1) developing new tailored treatment strategies with enhanced efficacy and reduced toxicity, (2) improving the patient selection approaches for these new treatments, and (3) advancing our understanding of how to harness the recent developments in immunotherapy to support other therapeutic strategies to prevent or overcome endocrine resistance and disease progression. It is our hope that continuing translational research will unveil more converging targets and pathways associated with altered ER transcriptional reprogramming, which can be therapeutically exploited to prevent and/or reverse endocrine resistance.

Purpose: To investigate the direct effect and therapeutic consequences of epidermal growth factor receptor 2 (HER2)-targeting therapy on expression of estrogen receptor (ER) and Bcl2 in preclinical models and clinical tumor samples. Experimental design: Archived xenograft tumors from two preclinical models (UACC812 and MCF7/HER2-18) treated with ER and HER2-targeting therapies and also HER2+ clinical breast cancer specimens collected in a lapatinib neoadjuvant trial (baseline and week 2 posttreatment) were used. Expression levels of ER and Bcl2 were evaluated by immunohistochemistry and Western blot analysis. The effects of Bcl2 and ER inhibition, by ABT-737 and fulvestrant, respectively, were tested in parental versus lapatinib-resistant UACC812 cells in vitro. Results: Expression of ER and Bcl2 was significantly increased in xenograft tumors with acquired resistance to anti-HER2 therapy compared with untreated tumors in both preclinical models (UACC812: ER P = 0.0014; Bcl2 P < 0.001 and MCF7/HER2-18: ER P = 0.0007; Bcl2 P = 0.0306). In the neoadjuvant clinical study, lapatinib treatment for 2 weeks was associated with parallel upregulation of ER and Bcl2 (Spearman coefficient: 0.70; P = 0.0002). Importantly, 18% of tumors originally ER-negative (ER−) converted to ER+ upon anti-HER2 therapy. In ER−/HER2+ MCF7/HER2-18 xenografts, ER reexpression was primarily observed in tumors responding to potent combination of anti-HER2 drugs. Estrogen deprivation added to this anti-HER2 regimen significantly delayed tumor progression (P = 0.018). In the UACC812 cells, fulvestrant, but not ABT-737, was able to completely inhibit anti–HER2-resistant growth (P < 0.0001). Conclusions: HER2 inhibition can enhance or restore ER expression with parallel Bcl2 upregulation, representing an ER-dependent survival mechanism potentially leading to anti-HER2 resistance. Clin Cancer Res; 21(17); 3995–4003. ©2015 AACR.

Background: The CDK4/6 inhibitors (i) palbociclib (Palbo), ribociclib, and abemaciclib remarkably improved the outcome of patients with metastatic ER+/HER2- breast cancer (BC) and are now under clinical investigation in early BC. Despite high efficacy, de novo and acquired resistance to CDK4/6i is common. Elucidating the molecular basis for sensitivity and resistance to CDK4/6i is crucial to identify predictive biomarkers and novel therapeutic targets to improve patient outcome. Materials and Methods: MCF7, T47D and ZR75-1 parental (P) BC cells and their derivatives made resistant to tamoxifen, estrogen deprivation (EDR), or fulvestrant were used. The P and EDR models of MCF7 and T47D cells were chronically exposed to increasing concentrations of Palbo to generate derivatives with acquired resistance to Palbo (PalboR). The transcriptomic profiles of P, endocrine-resistant (EndoR) and PalboR models were determined by RNA-seq. IC50s were determined by exposing MCF7, T47D, and ZR75-1 P and EndoR lines (n=12) to increasing concentrations of Palbo. Cell growth was assessed by methylene blue staining, and changes in the mRNA and protein levels of key cell cycle molecules were assessed by RT-PCR and Western blot, respectively. Gene expression data from the Cancer Dependency Map (DepMap), baseline tumors from the NeoPalAna (NCT01723774) and neoMONARCH (NCT02441946) neoadjuvant trials, as well as the TCGA and METABRIC datasets were interrogated for correlations of gene signatures and patient outcome (by KMPlot). Results: Palbo treatment resulted in a dose-dependent inhibition of the growth of P and EndoR BC cell lines, with varying degree of sensitivity among the models. GSEA analysis comparing the least sensitive (IC50>350nM) vs. the most sensitive (IC50 Citation Format: Carmine De Angelis, Xiaoyong Fu, Maria Letizia Cataldo, Agostina Nardone, Valerie M. Jansen, Jamunarani Veeraraghavan, Sarmistha Nanda, Lanfang Qin, Vidyalakshmi Sethunath, Resel Pereira, Matteo Benelli, Ilenia Migliaccio, Luca Malorni, Joshua Donaldson, Pier Selenica, David N. Brown, Britta Weigelt, Jorge S. Reis-Filho, Ben H. Park, Sara A. Hurvitz, Dennis J. Slamon, Mothaffar F. Rimawi, Rinath Jeselsohn, Kent Osborne, Rachel Schiff. High levels of interferon-response gene signatures are associated with de novo and acquired resistance to CDK4/6 inhibitors in ER+ breast cancer [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr GS2-01.

Background: The HER2 pathway can be inhibited by potent targeting agents such as lapatinib (L), trastuzumab (T), or their combination (LT), but acquired and de novo resistance still occur. Resistance to these drugs remains a major hurdle in the management of HER2+ breast cancer. Consequently, elucidation of mechanisms of acquired therapeutic resistance to HER2-directed therapies is of critical importance. Methods: To obtain clues to the mechanisms for resistance we developed a panel of HER2+ breast cancer cell lines resistant to L, T, or LT. Parental cells and resistant derivatives of the HER2+ BT474 cell line were characterized by RNA-seq. Genes that were overexpressed in resistant compared to parental cells were confirmed by RT-PCR, Western blotting, and immunohistochemistry (IHC). Cell growth and cell signaling were assessed in parental and resistant cell lines after down-regulation (by siRNA) or overexpression (via an inducible cDNA) of IRS4 in the presence or absence of treatment. The effect of IRS4 overexpression on L resistance was assessed in a BT474 xenograft model. The proteins that interact with IRS4 were identified by co-immunoprecipitation with IRS4 followed by separation of the associated proteins by SDS-PAGE and microsequencing by mass spectrometry. Results: RNA-seq analysis revealed that IRS4 was the most up-regulated gene in BT474 L or LT resistant derivatives in which HER2 signaling is effectively inhibited, but not T alone, where HER2 signaling is reactivated. Western blotting and IHC validated this result and identified membrane localization of IRS4. Knockdown of IRS4 in L- or LT-resistant cells reversed resistance and restored growth inhibition. IRS4 knockdown also inhibited downstream signaling, with a reduction in pAKT but not in pMAPK. Induction of the cell cycle regulator p27 and down-regulation of survivin were observed after IRS4 knockdown. Overexpression of IRS4 cDNA in parental BT474 and SKBR3 cells led to resistance to L/LT, increased pAkt, and decreased the apoptotic marker cleaved PARP in the presence of L or the LT combination. The BT474 xenograft model showed that IRS4 overexpression in the absence of treatment had no effect on tumor growth but it significantly reduced the inhibitory effect of lapatinib (p=0.002). A group of proteins that interact with IRS4 in BT474 L-resistant cells were identified by mass spectrometry. The roles of these proteins in IRS4-mediated resistance to lapatinb-containing regimens are under investigation. Conclusion: IRS4 overexpression is a critical factor in causing resistance to lapatinib-containing regimens in BT474 cells. Investigation of IRS4 and its signaling partners in HER2+ human tumors resistant to lapatinib will be important to determine if this mechanism is also operative in patients. Citation Format: Lanfang Qin, Maria B Hahn, Xiaoyong Fu, Martin J Shea, Mario Giuliano, Sarmistha Nanda, Xiaowei Xu, Huizhong Hu, Sung Yun Jung, Laura M Heiser, Nicholas Wang, Joe W Gray, Susan G Hilsenbeck, Chad Creighton, Chad A Shaw, Gary C Chamness, Dean P Edwards, Sabrina Herrera, Carolina Gutierrez, C Kent Osborne, Rachel Schiff. Overexpression of insulin receptor substrate 4 can mediate acquired resistance to lapatinib-containing regimens in HER2+ breast cancer cells [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P3-05-13.

Green supplier development has become necessary as organizations increasingly compete on environmental supply chain capabilities. However, formal modeling tools to aid managers in evaluating green supplier development programs (GSDPs) are virtually non-existent. This paper introduces a formal structured managerial approach for organizations to help evaluate the influence relationships amongst GSDPs. Utilizing GSDP categorizations we acquire multi-functional managerial inputs within a telecommunication systems provider to evaluate the GSDPs. The managerial inputs are evaluated using a formalized grey-based DEMATEL methodology. Managerial and research implications of the proposed study and application, limitations and future research directions are also detailed.

Multiple lines of preclinical and clinical evidence suggest a continued, albeit altered, role of estrogen receptor (ER) in endocrine resistance, a major hurdle in the clinical management of ER+ breast cancer. In the recent years, several studies that explored the biology of ER in recurrent metastatic disease in patients and in endocrine-resistant preclinical models have uncovered various mechanisms underlying altered ER activity and endocrine resistance. Some of these mechanisms are observed to emerge under the pressure of chronic endocrine therapy, through selective clonal expansion, resulting in acquired resistance. Recurrent gain-of-function mutations in the ligand-binding and other domains of the ER encoding gene ESR1 have been identified in 20-50% of the patients with endocrine-resistant metastatic breast cancers. Tumor biopsies have long been useful in identifying various mutations, but circulating tumor DNA in patients with advanced disease is now also emerging as a clinically valuable resource enabling the noninvasive detection of ESR1 mutations. Most of the ESR1 mutations confer constitutive ligand-independent ER activity, causing resistance to estrogen deprivation (i.e., aromatase inhibitors) and reduced sensitivity to the selective ER modulator (SERM) tamoxifen and the selective ER downregulator (SERD) fulvestrant. Less frequent ESR1-related genomic alterations, including gene amplifications, rearrangements, and gain-of-function gene fusions identified mostly in patients with recurrent metastatic disease, represent additional mechanisms of acquired endocrine resistance. While the therapeutic value of newly developed orally bioavailable SERMs/SERDs such as GDC-0810 and GDC-0927 for patients with tumors harboring ESR1 mutations is yet to be clinically proven, we anticipate that ongoing studies will address this open question. Additionally, the recent clinically potent CDK4/6 inhibitor palbociclib, in combination with fulvestrant, has shown promising therapeutic efficacy against tumors harboring ESR1 mutations. Acquired amplification of the aromatase-encoding gene CYP19A1, recently reported in ~20% of aromatase inhibitor-treated relapsed patients, has also been linked to endocrine resistance due to increased aromatase activity and intratumoral estrogen levels. Besides genomic alterations in ER, crosstalk between growth factor receptor (GFR) signaling pathways and ER can also mediate resistance by altering the activity of ER and its co-regulators, and as a result, the ER-dependent transcriptional program. In particular, the bidirectional crosstalk between ER and the HER family and other GFRs, including in ER+/HER2- tumors that harbor HER2 activating mutations and FGFR1 amplification in 10-15% of patients with ER+ breast cancer, has been implicated in endocrine resistance. Combined treatment with endocrine therapy plus inhibitors against pertinent GFRs or their downstream signaling components to circumvent resistance is currently under preclinical and clinical evaluation. Other factors that contribute to endocrine resistance are changes in ER-interacting/co-regulator proteins such as the ER co-activator SRC3, transcription factor complex AP-1, and the pioneer factor FOXA1. Most recently, a new smal- molecule inhibitor targeting SRC3 has been shown to inhibit the levels and/or transcriptional activity of the SRC family of coactivators and the growth of endocrine-naïve and resistant tumors in vivo. Clinical findings and preclinical data from in vivo mouse xenograft models have shown that the endocrine-resistant tumors harbor increased AP-1 activity, which mediates a shift in the ER transcriptional program and promotes endocrine resistance and metastatic capabilities. Numerous preclinical findings have long suggested the crucial role of the ER pioneer factor FOXA1 in cancer development and progression. In breast cancer, we first reported that high levels of FOXA1, by gene amplification and/or overexpression, promote acquired endocrine resistance across multiple endocrine-resistant cell and xenograft mouse models. In the clinical setting, recent sequencing studies have reported FOXA1 genomic aberrations, including gene amplification, recurrent mutations, and overexpression via activating mutations in the promoter region, in about 10% of ER+ primary tumors, and enrichment in metastatic tumors. Further, in the metastatic setting, FOXA1 genetic aberrations and ESR1 mutations associated with aromatase inhibitor resistance appear to be mutually exclusive, suggesting the involvement of different mechanisms in high FOXA1-induced endocrine resistance and metastasis. Our new studies in acquired endocrine-resistant preclinical models suggest that FOXA1 gene amplification and/or overexpression results in a global enhancer and transcriptional reprogramming that activates key downstream transcription factors, including the hypoxia-induced factor HIF2α, and the prometastatic secretome. Our findings thus support the notion that, at least in a subset of patients with ER+ breast cancer, high levels or activity of FOXA1 are drivers of endocrine resistance and metastasis, presenting new therapeutic targets and novel therapeutic approaches along its reprogrammed transcriptional axis at both the regulatory and downstream effector levels. These findings underscore the need for a more careful assessment of the genomic, epigenomic, and signaling landscape of recurrent lesions in order to devise more tailored therapies to effectively circumvent and/or prevent endocrine resistance and metastasis. In addition, a more complete suppression of the levels and activity of ER, its co-activators, and alternative survival pathways may prove to be a better therapeutic strategy in effectively combating endocrine resistance. Effective therapeutic strategies may include novel epigenomic approaches to inhibit gene expression and activity of ER and its co-activators, more potent SERDs, small-molecule inhibitors of SRC3 and other ER co-activators, signal transduction inhibitors that modulate levels and activity of ER and its co-activators, a new HIF2α small-molecule inhibitor currently under clinical evaluation in renal cell carcinoma, and development of drugs that specifically target the pioneer factor FOXA1. Citation Format: Rachel Schiff, Jamunarani Veeraraghavan, Xiaoyong Fu. Endocrine resistance in metastatic breast cancer: Mechanisms and new therapeutic strategies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr SY01-01.

ZrO2 films were prepared by electron beam evaporation with glancing angle deposition (GLAD) technique. The as-deposition and annealed ZrO2 films are all amorphous, different from that deposited at normal incidence. Due to the shadowing effect, a highly orientated structure composed of slanted columns formed, and the obtained films became the mixture of slanted columns and voids. The relationship among the effective refractive index, packing density and flux incident angle was investigated. The refractive index and packing density of ZrO2 films decrease with the increase of the incident angle. The in-plane birefringence of GLAD ZrO2 films was calculated. At the packing density of 0.576, the maximum birefringence is 0.037. (c) 2006 Elsevier B.V. All rights reserved.

The glancing angle deposition (GLAD) technique was used to deposit ZnS films by electron beam evaporation method. The cross sectional scanning electron microscopy (SEM) image illustrated a highly orientated microstructure composed of slanted column. The atomic force microscopy (APM) analysis indicated that incident flux angle had significant effects on the nodule size and surface roughness. Under identical nominal thickness, the actual thickness of the GLAD films is related to the incident flux angle. The refractive index and in-plane birefringence of the GLAD ZnS films were discussed, and the maximum bireffingence Delta n = 0.036 was obtained at incident flux angle of alpha = 80 degrees. Therefore, the glancing angle deposition technique is a promising way to create a columnar structure with enhanced birefringent property. (c) 2005 Elsevier B.V. All rights reserved.

Based on scalar diffraction theory, we investigated far-field intensity distribution (FFID) of beam generated by Gaussian mirror resonator. We found usable analytical expressions of diffracted field with respect to variation of diffraction parameters. Particular attention was paid to the parameters such as mirror spot size and radius of the Gaussian mirror, which determine the FFID. All analyses were limited to TEM00 fundamental mode. (c) 2004 Elsevier B.V. All rights reserved.

An all-fiber optic catheter-based polarization-sensitive optical coherence tomography system is demonstrated. A novel multiplexing method was used to illuminate the sample, splitting the light from a 58.5 kHz Fourier-domain mode-locked laser such that two different polarization states, alternated in time, are generated by two semiconductor optical amplifiers. A 2.3 mm forward-view cone-scanning catheter probe was designed, fabricated, and used to acquire sample scattering intensity and phase retardation images. The system was first verified with a quarter-wave plate and then by obtaining intensity and phase retardation images of high-birefringence plastic, human skin in vivo, and untreated and thermally ablated porcine myocardium ex vivo. The system can potentially in vivo image of the cardiac wall to aid radio-frequency ablation therapy for cardiac arrhythmias.

In this letter, a guided-mode resonance (GMR) filter with the same material for both the grating layer and the waveguide layer is designed, and its optical properties are investigated. The GMR filter owns almost 100% reflection at the resonance wavelength of 800 nm with the full-width at half-maximum (FWHM) of 20 nm, and its sideband reflection from 700 to 1000 nm is less than 5%. As the resonance wavelength is influenced by more than one parameter during the fabrication process, GMR filter with the same resonance wavelength can be obtained by adjusting other parameters or even one parameter to deviate from the design value.

An internal Brewster guided-mode resonance (GMR) filter is designed. For this kind of GMR filter, the Brewster reflection occurs at the interface of grating/waveguide layers rather than at the interface of air/grating layers. At Brewster angle of 60\circ the GMR filter owns almost 100% reflection at the resonance wavelength of 800 nm with the full-width at half-maximum (FWHM) of 0.2 nm. Its angle response changing with the fabrication deviation is also discussed.

The reflection and transmittance of s- and p-polarized lights should be treated separately at nonzero angle incidence. By choosing two special film materials whose refractive indices meet the Brewster condition in a given incident angle, the reflection of p-polarized light can be eliminated. When the two materials are fabricated into multilayer stacks, the reflection of s-polarized light can be largely increased while the reflection of p-polarized light keeps low. According to this principle, a film polarizer with bandwidth of 198 nm or even more is designed.

Introduction: The human epidermal growth factor receptor 2 (HER2)-targeted therapies trastuzumab (T) and lapatinib (L) show high efficacy in patients with HER2-positive breast cancer, but resistance is prevalent. Here we investigate resistance mechanisms to each drug alone, or to their combination using a large panel of HER2-positive cell lines made resistant to these drugs. Methods: Response to L + T treatment was characterized in a panel of 13 HER2-positive cell lines to identify lines that were de novo resistant. Acquired resistant lines were then established by long-term exposure to increasing drug concentrations. Levels and activity of HER2 and estrogen receptor (ER) pathways were determined by qRT-PCR, immunohistochemistry, and immunoblotting assays. Cell growth, proliferation, and apoptosis in parental cells and resistant derivatives were assessed in response to inhibition of HER or ER pathways, either pharmacologically (L, T, L + T, or fulvestrant) or by using siRNAs. Efficacy of combined endocrine and anti-HER2 therapies was studied in vivo using UACC-812 xenografts. Results: ER or its downstream products increased in four out of the five ER+/HER2+ lines, and was evident in one of the two intrinsically resistant lines. In UACC-812 and BT474 parental and resistant derivatives, HER2 inhibition by T reactivated HER network activity to promote resistance. T-resistant lines remained sensitive to HER2 inhibition by either L or HER2 siRNA. With more complete HER2 blockade, resistance to L-containing regimens required the activation of a redundant survival pathway, ER, which was up-regulated and promoted survival via various Bcl2 family members. These L-and L + T-resistant lines were responsive to fulvestrant and to ER siRNA. However, after prolonged treatment with L, but not L + T, BT474 cells switched from depending on ER as a survival pathway, to relying again on the HER network (increased HER2, HER3, and receptor ligands) to overcome L's effects. The combination of endocrine and L + T HER2-targeted therapies achieved complete tumor regression and prevented development of resistance in UACC-812 xenografts. Conclusions: Combined L + T treatment provides a more complete and stable inhibition of the HER network. With sustained HER2 inhibition, ER functions as a key escape/survival pathway in ER-positive/HER2-positive cells. Complete blockade of the HER network, together with ER inhibition, may provide optimal therapy in selected patients.

Background: Poly [ADP-ribose] polymerase (PARP) is an important mediator of DNA damage repair. Preclinical and clinical studies have indicated PARP inhibitors (i) as promising agents in DNA-repair-defective cancers, especially in the presence of BRCA1/2 alterations. Recent evidence also suggests a role for additional DNA damage-independent functions of PARP, involving transcriptional and epigenetic regulation. Particularly, PARP-1 can interact with the estrogen receptor (ER) and modulate its transcriptional activity in vascular smooth muscle cells, and PARPi can circumvent endocrine resistance in prostate cancer cells. However, the effect of PARPi on the efficacy of endocrine therapy (EndoTx) in ER+ breast cancer is unknown. Here we aimed to establish these effects using the PARPi olaparib (AZD2281, Olap) in our preclinical models. Methods: The effects of Olap (0.01-1μM) alone or in combination with EndoTx [estrogen deprivation (ED) or tamoxifen (Tam)] were tested in vitro in three ER+ parental cell lines (MCF7, ZR75-1, and T47D; BRCAwt) and their endocrine resistant (R) derivatives. Clonogenic assays were used to assess Olap-induced changes in cell growth. In vivo, ovariectomized nude mice bearing 200 mm3 tumors growing in the presence of estrogen supplementation (E2 pellets), were randomized to either continued E2 (control), ED, or ED+Tam; all ± oral Olap (100mg/kg daily). Results: In MCF7 parental cells in vitro, Olap, in a dose dependent manner, inhibited E2-stimulated growth and increased the growth inhibitory effect of EndoTx. In addition, at a clinically relevant dose (1μM), Olap significantly inhibited the growth of MCF7 EndoR cell derivatives. Similar results were also observed in the other 2 ER+ cell models. In vivo, long-term Olap-treatment in the presence of EndoTx (Tam ∼200days, and ED ∼300days) was well tolerated and no apparent drug-related toxicity was observed. Olap did not affect the growth of E2-stimulated MCF7 xenografts. In contrast, Olap enhanced sensitivity to Tam by delaying time to tumor progression (TTP; size doubling) from a median of 95 days to 155 days (p = 0.03), and numerically but not significantly reduced time to tumor regression (TTR; size halving) from 138 to 51 days (p = 0.2). Olap also numerically accelerated median time to tumor regression in the presence of ED (34 ED vs. 25 days ED+Olap; p = 0.23), but had no effect on TTP (P = 0.36). Conclusion: Our results suggest a potential role for Olap in enhancing EndoTx efficacy and circumventing resistance in the absence of BRCA mutations. The absence of any toxicity and negative interaction by adding Olap to EndoTx in the in vivo experiment supports the inclusion of ER+ breast cancer patients in clinical trials using PARPi. Further studies are warranted to better understand the biology of PARP and the efficacy of PARPi in ER+ breast cancer, especially in the presence of EndoTx and resistance. Citation Format: Agostina Nardone, Amit Goldstein, Martin J. Shea, Tamika Mithchell, Xiaoyong Fu, Carmine De Angelis, Huizhong Hu, Xiaowei Xu, Mahitha Rajendran, Mark O'Connor, Gershon Locker, Susan Hilsenbeck, Kent Osborne, Rachel Schiff. PARP inhibition effects on endocrine therapy and resistance in estrogen receptor positive (ER+) breast cancer models. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr LB-166. doi:10.1158/1538-7445.AM2015-LB-166

Glancing angle deposition (GLAD) is a novel way to produce nanostructural thin films with engineered porosity, and it is possible to make new optical components in laser systems. In this paper, ZrO 2 , SiO 2 and TiO 2 thin films were grown by electron beam evaporation with GLAD technique. Different mi crostructures were observed. The optical properties, such as transmittance and refractive index were characterized. As application of the GLAD thin films, several optical components were designed and fabricated, such as graded-index rugate filter, br oadband antireflection coating and phase retardater for visible and near infrared laser systems. Finally, laser-induced damage threshold were measured and discussed. Key Words: thin films; glancing angle deposition; nanostructure; optical component; laser-induced damage threshold 1. INTRODUCTION Recently, glancing angle deposition (GLAD) technique has attracted considerable attention for de signing the novel morphology of the nanostructured films [1, 2]. This technique employs substrate inclination and substrate rotation, which were driven by two step motors controlled by a computerized program [3, 4]. As a result of enhanced shadowing effect and limited atom diffusion during film growth, the GLAD thin films have high porosity and anisotropic growth behavior . GLAD technique has been shown to be capable of producing films with various microstructures, such as vertical columns [5], helix [6], which is hardly obtained from the normal incidence vapor deposition. Thus, many novel physical and chemical properties of thin films can also be easily engineered. In this paper, ZrO

Free-space optical transmission can provide high-speed links for a variety of applications. However, inhomogeneities in the temperature and pressure of the atmosphere lead to variations of the refractive index along the transmission path. This atmospheric turbulence can deteriorate the quality of the image formed at the receiver, and can cause fluctuations in both the intensity and the phase of the received signal. Laser beams experience three effects under turbulence: beam wander, beam spreading and scintillation. Of the three turbulence effects, free space optical systems might be most affected by scintillation. Random interference with the wave front can cause peaks and dips, resulting in receiver saturation or signal loss. Based on Kolmogorov’s theory, the Signal-to-Noise Ratio (SNR) and Bit Error Rate (BER) from scintillation were drawn in this paper. For a FSO system, after choosing a certain beam and communication range, link performance characterized by its SNR or BER is related to the refractive index structure coefficient C n 2 and turbulence depth L t . Considering all the three effects, we find for a weak turbulence model, a FSO system should be applied within a communication range of 2.5km and above the ground at least 5m.

Background: We have recently shown that the luminal B subtype breast cancer (BC) is associated with PI3K molecular signature. Breast tumors of the luminal B subtype generally have lower ER expression levels and/or activity and are more aggressive and less sensitive to endocrine therapy when compared to the luminal A tumors. To further investigate the role of PI3K signaling in BC luminal types and endocrine resistance, we have characterized an array of luminal A and B cell lines, as well as their endocrine resistant derivatives, to examine how modulation of the PI3K pathway can influence ER activity and endocrine sensitivity. Materials and Methods: We focused on endocrine resistant derivatives of two luminal A (ZR75-1 and BT483) and two luminal B (ZR75-B and CAMA-1) cell lines. Resistant clones were developed by long-term (>6 months) endocrine treatment [estrogen deprivation (ED) or ED plus Tamoxifen (Tam)]. IC50 values of the PI3K inhibitor BEZ-235 were determined in the parental and the endocrine resistant lines, and qRT-PCR was performed to measure the effect of the inhibitor on mRNA levels of ER and its regulated genes (e.g. PR, CAV1, IGF1R and IRS1). Cells were also treated (4 days) with Tam, BEZ-235, or the combination and growth inhibition assays were conducted to assess the drugs efficacy. Inducible PTEN knockdown clones were established in MCF7L and BT483 cell lines. Endocrine sensitivity of these clones was studied in vitro and in vivo, using xenograft mouse models. Results: Ten days of treatment showed that all parental lines were sensitive to endocrine therapy. In contrast, stabilized endocrine resistant cell lines showed similar cell growth rate under Tam or ED treatment compared to control E2 treatment (p>0.05). ZR75-1 endocrine resistant derivatives acquired higher sensitivity to BEZ-235 treatment compared to the parental (p Discussion: The inverse association between PI3K and ER pathways in luminal BC cell lines validates the relevance of these models for further studies. Endocrine resistant phenotype may be partially caused by activated PI3K signaling. Our data suggest that inhibition of PI3K signaling can restore ER levels and/or activity, thereby reversing the aggressive and hormonal-refractory phenotype of the tumors, and achieving improved endocrine therapy response. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2290. doi:10.1158/1538-7445.AM2011-2290

A guided-mode resonance (GMR) filter with the same material (Ta2O5) for both the grating layer and the waveguide layer is designed and fabricated. This simple structure is easy to fabricate and can avoid the defects at the grating/waveguide interface using different materials. The spectral response measured with a Lambda 900 spectrophotometer under normal incidence for TE waves exhibits a peak reflectance exceeding 80% at the wavelength of 1040 nm with a full-width half-maximum (FWHM) linewidth of 23 nm. We evaluate the deviations of the fabricated structure from the designed parameters.

A normal-incidence nonpolarizing guided-mode resonance filter is designed. There are two waveguide layers and one grating layer in the filter. By adjusting the distance between the two waveguide layers, the same resonance wavelength for both TE and TM polarization can be achieved. An antireflection design method is also used to decrease the sideband reflection of the filter. The results show that the filter has high reflection, more than 99.9% at 500 nm, and the FWHMs of TE- and TM-polarized light are 2.16 and 0.15 nm, respectively.

The far-field intensity distribution of hollow Gaussian beams was investigated based on scalar diffraction theory. An analytical expression of the M2 factor of the beams was derived on the basis of the second-order moments. Moreover, numerical examples to illustrate our analytical results are given.