bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P4-14-18: Phase I study of single-agent pyrotinib, a novel irreversible pan-ErbB receptor tyrosine kinase inhibitor, in patients with ErbB2+ metastatic breast cancer
Authors: Xu B , Ma F , Chen S , Li Q , Yang F , Zhang Y , Chen X , Zhong D , Zhang G
Published: May 2016

Pyrotinib is a novel small molecule irreversible pan-ErbB receptor tyrosine kinase inhibitor. This phase I study was designed to evaluate the safety, pharmacokinetics and antitumor activity of pyrotinib in Chinese patients with ErbB2+ metastatic breast cancer (n=38). The majority of patients were previously treated with multiple lines (≤5) of anti-tumor regimens including trastuzumab. The dose-escalation study was conducted at 80, 160, 240, 320, 400 and 480 mg levels in patients receiving oral pyrotinib with food once daily on a 28-day cycle. Dose-limiting toxicity (DLT) was grade 3 diarrhea, which occurred in two out of two patients treated with 480mg pyrotinib during the dose-escalation. Therefore, the maximum tolerated dose (MTD) of pyrotinib was determined to be 400 mg per day. In general, pyrotinib was safe and well tolerated when administered to patients. The majority of adverse events which occurred in the study were grade I and II. The pharmacokinetic results showed linear characteristics along the range of doses tested, and supports a once daily dosing regimen. The overall objective response rate (ORR) was 52.8% and disease control rate (DCR) was 80.6% in 36 patients (19 PR and 10 SD), with the highest ORR of 55.6% (5/9) and 87.5% (7/8) observed in 320 mg and 400 mg cohorts, respectively. The median duration of response was 32.3 weeks. In addition, 3 (8.3%) subjects achieved stable disease for at least 24 weeks. Kaplan-Meier median progression-free survival (PFS) was 35.3 weeks (95%CI: 23.6∼39.9 weeks) among all subjects treated with 80-400 mg, and the median PFS for 320 mg and 400 mg cohorts was 39.9 weeks (95%CI: 31.1∼47.4 weeks). Based on tolerability, safety, PK and efficacy data of 6 dose-cohorts, 320 mg and 400mg of pyrotinib once daily could be considered a recommended regimen for Phase II studies. The promising antitumor activity of pyrotinib observed in patients with ErbB2+ metastatic breast cancer, who had previous treatment with anthracyclines, taxanes, and trastuzumab, warrants its further study (ClinicalTrials.gov number, NCT01937689).

Citation Format: Xu B, Ma F, Chen S, Li Q, Yang F, Zhang Y, Chen X, Zhong D, Zhang G. Phase I study of single-agent pyrotinib, a novel irreversible pan-ErbB receptor tyrosine kinase inhibitor, in patients with ErbB2+ metastatic breast cancer. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P4-14-18.
Abstract A23: Fe en Acción/Faith in Action: Promotion of cancer screening among churchgoing Latinas
Authors: Elder John P. , Arredondo Elva M. , Haughton Jessica , Perez Lilian G. , Martínez María Elena
Published: May 2016

Objectives: To present outcomes of a controlled two-group clustered randomized community trial promoting breast, cervical, skin and colorectal cancer screening (CS) among churchgoing Latinas through promotoras (lay health advisors).

Design: Sixteen churches were randomly assigned either to the CS or a physical activity (PA) condition. Sixteen promotoras at the 8 CS churches intervened at the individual- (knowledge and beliefs), interpersonal- (social support and sharing personal experiences), organizational- (space at church for program activities), and environmental- (access to health care) levels to affect changes in target behaviors. Approximately 27 women per church were recruited to participate in the study.

Measurements: The study's primary outcome was being current on recommended mammography, clinical breast exams, sigmoidoscopy and colonoscopy, fecal occult blood test, and cervical cancer tests based on self-report at baseline and 12 and 24 months following implementation of intervention activities. Participants were 436 Latinas (aged 18-65 years; 91% Mexican immigrants) who met activity-level criteria for the PA condition, attended church at least four times per month, and lived near their church. Enrolled participants were asked to complete measures assessing PA and cancer screening as well as their correlates at each measurement point (e.g., health insurance, perceived barriers to screening, and knowledge about each cancer).

Results: The CS intervention had a significant positive impact on self-reported uptake of mammography and breast exams. Pap smear tests were already reported at high baseline rates and did not change as a function of the intervention. Colorectal cancer screening rates were low and did not improve with the intervention.

Summary: Promotoras working in their own churches can have a significant impact on promoting breast cancer screening among Latinas. The use of colon cancer screening remains a difficult challenge, however, with improved methods still being needed.

Citation Format: John P. Elder, Elva M. Arredondo, Jessica Haughton, Lilian G. Perez, María Elena Martínez. Fe en Acción/Faith in Action: Promotion of cancer screening among churchgoing Latinas. [abstract]. In: Proceedings of the Eighth AACR Conference on The Science of Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; Nov 13-16, 2015; Atlanta, GA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2016;25(3 Suppl):Abstract nr A23.
Abstract A44: Using CBPR to extend prostate cancer education, counseling, and screening opportunities to urban-dwelling African Americans
Authors: Ross Levi , Johnson Jarrett , Smallwood Stacy W. , Luque John (Juan) , Tedders Stuart H. , Airhihenbuwa Collins O. ,   MAN UP Prostate Cancer Advocates
Published: May 2016

Community-based participatory research (CBPR) is becoming one of the dominant approaches for bringing evidence- and consensus-based cancer prevention and control practices to medically underserved communities. There are many examples of how CBPR has been useful for generating culturally specific solutions for different health issues that affect African-Americans. However, few examples exist in the literature on how the CBPR approach can be applied to address prostate cancer. This presentation will describe a collaborative process for linking inner-city, African-American men to free prostate cancer education, physician counseling, and screening opportunities (prostate-specific antigen (PSA) testing and digital rectal examination (DRE)). The site of this community-based participatory project was the city of Buffalo, located in Erie County, New York. A total of 55 men from 16 zip codes attended the three hour community event. All 55 attendees received free prostate cancer education, and 35 (63%) chose to undergo screening. Of the 35 men who were screened, the majority (29/35= 83 %) were African-American. Event planners were pleased that nearly three fourths of those screened (26/35=74 %) were first-time screeners. All event attendees (55/55=100%), even those who elected not to screen, had an informed screening discussion with one of the attending urologists. All participants were mailed their screening results within 2 weeks, along with personalized instructions for following up with their primary care physician.The collaborative, community-academic process that will be described includes the following: (1) planning and conducting a community needs assessment to contextualize local prostate cancer issues, (2) organizing town and gown event planning, and (3) manipulating aspects of the built environment to build an infrastructure within the community to address disparities in screening opportunities. This presentation will conclude with a description of lessons learned that can help others develop and implement similar activities in other communities.

Citation Format: Levi Ross, Jarrett Johnson, Stacy W. Smallwood, John (Juan) Luque, Stuart H. Tedders, Collins O. Airhihenbuwa, MAN UP Prostate Cancer Advocates. Using CBPR to extend prostate cancer education, counseling, and screening opportunities to urban-dwelling African Americans. [abstract]. In: Proceedings of the Eighth AACR Conference on The Science of Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; Nov 13-16, 2015; Atlanta, GA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2016;25(3 Suppl):Abstract nr A44.
Abstract B90: Predictors of HPV vaccine initiation and completion among Hispanic mothers of 11-17 year old daughters living along the Texas-Mexico border
Authors: Morales-Campos Daisy Y. , Parra-Medina Deborah A.
Published: May 2016

Background: Cervical cancer incidence and mortality are higher for Hispanic women along the Texas-Mexico border than for other female population groups. Incidence could be reduced if teenaged Hispanic girls received the HPV vaccine before they became sexually active. However, few Hispanic girls compared to U.S. girls receive all three HPV vaccine doses (31% vs 36%), which prevent cervical cancer. Mothers are crucial to the success of HPV vaccine uptake efforts, but few studies have examined predictors of vaccine initiation and completion. The purpose of this study is to assess predictors (socio-demographics, HPV and vaccine knowledge, and vaccine self-efficacy) of vaccine initiation and completion.

Methods: We utilized baseline data from an outreach and education program utilizing promotoras and peer educators to deliver health education to mothers and daughters to increase HPV knowledge and promote HPV immunization. Our analyses utilized data from mothers of never vaccinated girls (n=371) to examine the association between the predictors and vaccine initiation and completion. To control for potential confounders, we conducted multivariable logistic regression analyses.

Results: Findings showed both health insurance (Initiators AOR: 0.27; 95% CI= .09, .79; Completers AOR: 0.29; 95% CI= .10, .83) and program status (Initiators AOR: 3.04; 95% CI= 1.76, 5.26; Completers AOR: 2.21; 95% CI= 1.27, 3.58) were associated with HPV vaccine initiation and completion among mothers. Results also showed preferred language (AOR: 0.15; 95% CI= .04, .53) and health status (AOR: 0.52; 95% CI= .29, .92) were associated with series initiation.

Conclusions: Findings suggest mothers who spoke English, had health insurance, and reported having good health were at lower odds of vaccine initiation. Also, mothers with health insurance also had lower odds of series completion. Mothers who participated in the program compared to the brochure only group were more likely to initiate and complete the vaccine series. These findings assist in identifying appropriate intervention strategies to enhance vaccine initiation in border communities at risk for cervical cancer.

Citation Format: Daisy Y. Morales-Campos, Deborah A. Parra-Medina. Predictors of HPV vaccine initiation and completion among Hispanic mothers of 11-17 year old daughters living along the Texas-Mexico border. [abstract]. In: Proceedings of the Eighth AACR Conference on The Science of Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; Nov 13-16, 2015; Atlanta, GA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2016;25(3 Suppl):Abstract nr B90.
Abstract C28: Racial and ethnic disparities in mammography utilization among women with intellectual disabilities: An exploratory mixed methods approach
Authors: Arana Evelyn
Published: May 2016

Background: Racial and ethnic disparities in mammography utilization among the general population persist to this day. Women with intellectual disabilities have much lower rates of mammography receipt compared to women without ID and are diagnosed with breast cancer at more advanced stages. Among women with ID, the diverse perceptions and experiences among different racial and ethnic groups surrounding mammography utilization are unknown. The purpose of this study is to conduct a mixed methods study to explore racial and ethnic differences in mammography utilization and experience among women with ID who are Black/African American, Hispanic, and non-Hispanic White.

Specific Aims:

1. Collect mammography medical records and demographic information from Black/African American, Hispanic, and non-Hispanic White women with ID to determine general trends in mammography utilization from 2010-2013.

2. Determine the barriers and facilitators contributing to mammography utilization among different racial and ethnic groups of this population via in-depth interviews.

3. Explore how culture may affect the social interactions that in turn may affect mammography utilization among this population.

This approach is innovative as it combines quantitative and qualitative methodologies to data collection among a priority population. The proposed research is significant because it will increase awareness on the different racial and ethnic perspectives regarding mammography utilization among women with ID. In the long-term, the results from this research may improve the delivery of appropriate healthcare to this population to achieve a reduction in mammography disparities. In addition, expected results from this research can be considered when planning breast cancer prevention interventions within this population and may be used to enhance the quality of healthcare for women with ID. This research is funded by AHRQ GRANT11714050. Preliminary findings are expected for November 2015.

Citation Format: Evelyn Arana. Racial and ethnic disparities in mammography utilization among women with intellectual disabilities: An exploratory mixed methods approach. [abstract]. In: Proceedings of the Eighth AACR Conference on The Science of Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; Nov 13-16, 2015; Atlanta, GA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2016;25(3 Suppl):Abstract nr C28.
Abstract A10: Oncogenomic analysis of IGF-IR driven mammary tumors identifies a potential cell of origin and mechanisms of tumor recurrence
Authors: Jones Robert A. , Campbell Craig I. , Watson Katrina L. , Moorehead Roger A.
Published: May 2016

Genetically engineered mice are valuable tools for understanding the genetic events that contribute to breast cancer development and progression. We have previously generated a doxycycline (Dox) inducible model of breast cancer, MTB-IGFIR, in which overexpression of the human type-I IGF receptor (IGF-IR) leads to the rapid induction of ER-mammary tumors with mixed luminal and basal features. Following Dox withdrawal and subsequent transgene downregulation, a subset of tumors escape IGF-IR dependence and spontaneously recur with evidence of an epithelial-mesenchymal transition (EMT). To gain insight into the molecular pathways that drive primary and recurrent tumor growth in MTB-IGFIR mice, we carried out gene expression profiling and DNA copy number analysis. Gene-set enrichment analysis (GSEA) revealed expression of an Akt oncogenic signature in primary tumors, consistent with downstream signaling mediated by IGF-IR. Interestingly, primary tumors also expressed a luminal progenitor gene signature suggesting a potential cell of origin. Accordingly, flow cytometric analysis of Dox-pulsed mice showed that high expression of the IGF-IR transgene was confined to EpCAM+/high Sca-1− cells, a putative ER- alveolar progenitor population. Recurrent tumors were found to express a mammary stem cell (MaSC) gene signature, which is consistent with their more mesenchymal phenotype as well as increased cell cycle pathway activity. While aCGH analysis identified few genomic changes in primary tumors, recurrent tumors exhibited gains and losses in known oncogenes and tumor suppressors including MET receptor, YAP1 and p16Ink4a/p19ARF. Integration of mRNA and copy number data confirmed altered expression of these genes at the transcriptional level. Taken together, these results suggest IGF-IR drives tumor initiation in an ER- luminal progenitor cell population in part through activation of the Akt pathway and that multiple genetic alterations contribute to the acquisition of oncogene independence and tumor relapse.

Citation Format: Robert A. Jones, Craig I. Campbell, Katrina L. Watson, Roger A. Moorehead. Oncogenomic analysis of IGF-IR driven mammary tumors identifies a potential cell of origin and mechanisms of tumor recurrence. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research; Oct 17-20, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(2_Suppl):Abstract nr A10.
Abstract A51: Investigation of the DEK oncogene as a blood biomarker for breast cancer
Authors: Sendilnathan Arun , Charif Mahmoud , Shaughnessy Elizabeth , Lewis Jaime D. , Radhakrishnan Neetu , Lower Elyse , Lanigan Catherine , Kumar Harriet , Wise-Draper Trisha M. , Vinnedge Lisa Privette
Published: May 2016

Breast cancer (BC) is one of the leading causes of death for women in the United States. This is due, in part, because women wait too long to get a diagnosis or the disease returns, sometimes years later, due to ineffective treatment. Therefore, it is critically important to identify biomarkers that can better inform clinicians while providing a screening test more amenable to patients than mammograms. We are investigating the feasibility of using DEK protein levels in patient plasma as a biomarker for disease stage and prognosis. DEK is a chromatin-organizing protein that functions in DNA replication and repair, transcription, and mRNA splicing. The DEK oncoprotein is highly upregulated in many types of cancer including breast, head and neck, and melanoma. Using human cell culture and murine models, our previous studies have shown that DEK is functionally important for the promotion of cellular growth, invasion/metastasis, drug resistance, and breast cancer stem cell maintenance. Work by the Markovitz laboratory indicated that DEK is secreted by macrophages as a pro-inflammatory signaling molecule and can be internalized by neighboring cancer cells to promote typical DEK functions. This led to our hypothesis that DEK may be present in the plasma of cancer patients. In fact, the detection of DEK protein in urine is currently being explored as a diagnostic biomarker for bladder cancer. To test this hypothesis, we collected peripheral blood from patients with newly diagnosed, untreated BC (irrespective of the clinical stage) and age- and ethnicity-matched normal healthy controls. Plasma was separated from the samples and subjected to DEK specific ELISA (Cusabio, Wuhan, China). Preliminary results demonstrate that DEK is indeed present in the plasma of BC patients and we are currently comparing these levels to normal healthy controls. Further analyses are ongoing to determine whether DEK levels correlate with pertinent clinical and pathological variables including age, tumor stage, and ER/PR/HER-2 status. In addition, we are examining if DEK levels can predict response to various treatment modalities and risk of relapse. These data will be important to verify DEK plasma measurements as a clinically useful test and may give insight to future personalized and targeted treatment strategies for BC.

Citation Format: Arun Sendilnathan, Mahmoud Charif, Elizabeth Shaughnessy, Jaime D. Lewis, Neetu Radhakrishnan, Elyse Lower, Catherine Lanigan, Harriet Kumar, Trisha M. Wise-Draper, Lisa Privette Vinnedge. Investigation of the DEK oncogene as a blood biomarker for breast cancer. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research; Oct 17-20, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(2_Suppl):Abstract nr A51.
Abstract B23: Citral reduces ALDH1A3 activity in breast cancer: Potential applications in targeting breast cancer stem cells
Authors: Thomas Margaret L. , Coyle Krysta M. , Cruickshank Brianne , Giacomantonio Michael , Wallace Melissa , Giacomantonio Carman , Marcato Paola
Published: May 2016

Breast tumors contain a subpopulation of cells known as cancer stem cells (CSCs) which are highly tumorigenic, resistant to typical chemo- and radiotherapy, and express high levels of aldehyde dehydrogenase (ALDH) isoforms ALDH1A3 and ALDH1A1. These enzymes initiate retinoic acid (RA) signaling and potentially contribute to detoxification of chemotherapy. A compound that inhibits these enzymes could be used as an adjuvant therapy to target breast CSCs and/or re-sensitize this population to chemotherapy. In particular, there are no currently characterized inhibitors of the ALDH1A3 isoform, so twelve general ALDH inhibitors were tested for their effectiveness in targeting ALDH1A3 activity. Ability to directly inhibit ALDH activity was quantified with the Aldefluor assay on a patient tumor xenograft and on breast cancer cell lines MDA-MB-231, MDA-MB-468, and SKBR3. Inhibition of RA signaling was quantified by measuring RA-inducible gene expression after 24 hours treatment with ALDH inhibitor. To investigate the anti-cancer properties of these compounds, apoptosis was quantified with the annexin V assay after 24 hours treatment. To determine if increased ALDH1A3 expression is involved in chemoresistance, MDA-MB-231 cells overexpressing ALDH1A3 and MDA-MB-468 cells with ALDH1A3 shRNA knockdown were treated for 72 hours with doxorubicin, 4-hydroperoxycyclophosphamide, or paclitaxel. Increased ALDH1A3 expression did not affect the amount of chemotherapy-induced apoptosis or cell proliferation rates. Citral significantly reduced ALDH1A3-mediated expression of RA-inducible genes and significantly reduced the ALDH activity of the patient-derived xenograft as well as ALDH1A3-overexpression MDA-MB-231 cells. Citral induced apoptosis in an ALDH1A3-dependant manner in breast cancer cell lines, and its ability to reduce MDA-MB-231 in vivo growth was investigated by implanting MDA-MB-231 cells in female NOD/SCID mice and injecting with 0.1mg/kg citral. Tumor size was not affected by citral injections, however recent data suggests that micelle-encapsulated citral has the potential to reduce tumor bulk. Should citral prove to be an effective breast cancer and ALDH1A3 inhibitor in future tumor xenograft experiments, this compound could potentially be used as adjuvant therapy for breast cancer patients.

Citation Format: Margaret L. Thomas, Krysta M. Coyle, Brianne Cruickshank, Michael Giacomantonio, Melissa Wallace, Carman Giacomantonio, Paola Marcato. Citral reduces ALDH1A3 activity in breast cancer: Potential applications in targeting breast cancer stem cells. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research; Oct 17-20, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(2_Suppl):Abstract nr B23.
Abstract B41: HGFL-Ron signaling enhances breast cancer stem cell populations
Authors: Ruiz-Torres Sasha J. , Waltz Susan E.
Published: May 2016

The Ron receptor tyrosine kinase is overexpressed in a high fraction of human breast cancers (BCs). Ron and its ligand, the Hepatocyte Growth Factor-Like protein (HGFL), play important roles during BC initiation, progression, and therapeutic resistance, with overexpression of this signaling cascade associated with increased BC metastasis and poor prognosis in humans. Reports demonstrate that a small subpopulation of cancer cells, called the breast cancer stem cells (BCSCs), are essential for the development and maintenance of BC, leading to tumor formation and recurrence. Despite the role of HGFL-Ron signaling as a key regulator of BC, nothing is known about this signaling pathway in BCSC populations. To investigate the function of HGFL-Ron signaling in BCSCs, the expression levels of Ron and HGFL were analyzed in BCSCs. mRNA analyses show increased levels of Ron and HGFL in BCSC-enriched Mammospheres (MSs) compared to parental BC epithelial cells. Next, the association between Ron signaling and the BCSC population was examined. Aldefluor assays of several Ron replete and depleted human and murine BC epithelial cell lines show that Ron overexpression increases the BCSC population. In vivo, flow cytometry analyses of breast tumors for BCSC markers show that Ron signaling is associated with increases in the BCSC population. MS formation assays using several Ron replete and depleted human and murine BC cell lines also demonstrate that cells expressing high levels of Ron have increased MS formation. Ex vivo studies of sorted BCSCs further support that Ron signaling enhances the MS formation ability of BCSCs. Interestingly, our data suggest that both genetic ablation and chemical inhibition of Ron signaling is sufficient to decrease BCSC MS formation ability. Mammosphere culture over several passages also suggest that Ron signaling BCSCs have higher self-renewal capabilities than Ron deficient BCSCs, highlighting the role of HGFL-Ron signaling in regulating BCSC function. Recent preliminary studies correlate Ron signaling in BCSCs with increased β-catenin and NF-κB pathways, suggesting these as the potential molecular mechanisms through which Ron signaling promotes BCSC function in BC. Overall, our data provide (i) the first evidence demonstrating both HGFL and Ron as critical factors regulating the BCSC population and (ii) that this regulation might occur in a β-catenin and NF-κB pathway dependent manner. These studies suggest both Ron and HGFL as potential therapeutic targets to eradicate BCSCs and improve the outcome of BC patients.

Citation Format: Sasha J. Ruiz-Torres, Susan E. Waltz. HGFL-Ron signaling enhances breast cancer stem cell populations. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research; Oct 17-20, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(2_Suppl):Abstract nr B41.
Abstract B48: Intratumor ploidy heterogeneity as a chemoresistance mechanism in triple-negative breastcancer
Authors: Gyanchandani Rekha , Lee Adrian
Published: May 2016

Background: Triple negative breast cancers (TNBC) are aggressive, heterogeneous tumors, and currently lack targeted therapies. Chemotherapy is the mainstay treatment, but tumor recurrence is a major cause of mortality and morbidity. Chemotherapy may select for resistant subclones that have a survival advantage over the bulk population of cancer cells and cause tumor relapse. DNA aneuploidy (abnormal number of chromosomes) is a common genetic feature in breast cancer and has been frequently associated with poor prognosis. Although, aneuploid or polyploid cancer cells can contribute to intratumor ploidy heterogeneity, their functional role in tumorigenesis and chemoresistance is largely undefined. Traditionally, polyploid cells were thought to arise due to defects in cell division and were considered to be senescent. However, recent reports have shown that polyploid cells represent a viable and proliferating subpopulation within a tumor and may have a direct role in regulating chemoresistance.

Methods: In this study, we isolated and functionally characterized the diploid and polyploid sub-populations derived from TNBC cell lines. DNA ploidy was assessed in Hoechst 33342 stained cells using flow cytometry. Cells with greater than 4N DNA content were considered as polyploid. Next, we analyzed cell proliferation in HCC1937 cells using CFSE and DNA labeling. Live-cell imaging was performed on flow-sorted diploid and polyploid cells using IncuCyte Zoom to monitor cell proliferation in the absence and presence of mitotic inhibitor (docetaxel).

Results: DNA ploidy analysis identified presence of polyploid cells in HCC1395 and HCC1937 cell lines at a frequency of 6.7% and 5.8% respectively. Polyploid cells have higher forward-scatter than diploid cells suggesting that they are larger in size. In addition, nuclear staining with Hoechst 33342 showed that giant polyploid cells contain enlarged and/or multiple nuclei. Time-lapse microscopy in polyploid cells revealed mitotic structures suggestive of multipolar cell division. Carboxyﬂuorescein diacetate, succinimidyl ester (CFSE) labeling showed that polyploid cycle slower than the predominant diploid population. Further, the IncuCyte cell proliferation assay also confirmed that sorted polyploid cells divide slower than the diploid cells and displayed resistance to docetaxel.

Concusions: These findings indicate that there are functional differences between diploid and polyploid sub-populations. In our future studies, we will examine the isolated diploid and polyploid cells for differences in their tumorigenic potential and role in tumor recurrence.

Citation Format: Rekha Gyanchandani, Adrian Lee. Intratumor ploidy heterogeneity as a chemoresistance mechanism in triple-negative breastcancer. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research; Oct 17-20, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(2_Suppl):Abstract nr B48.
Abstract B56: Prolactin promotes breast cancer to bone metastasis and breast cancer cell-mediated osteoclast differentiation
Authors: Forsyth Amanda , Sutherland Ashley , Cong Yingying , Grant Laurel , Juan Tzu-Hua , Lee Jae K. , Klimowicz Alexander , Petrillo Stephanie K. , Hu Jinghui , Chan Angela , Boutillon Florence , Goffin Vincent , Egan Cay , Tang Patricia A. , Cai Li , Morris Don , Magliocco Anthony , Shemanko Carrie S.
Published: May 2016

The hormones prolactin (PRL), estrogen and progesterone have long been studied for their role in the primary breast tumor but not yet in modulating the secondary tumor microenvironment of the bone. Metastasis to the bone is a debilitating aspect of many cancers, including breast cancer, where it is a preferred site of metastasis that results in bone loss. Breast cancer cells release osteolytic factors that induce the breakdown of bone, which releases growth factors and calcium that create a vicious cycle of metastatic tumor growth. Using quantitative immunohistochemistry (AQUA) (n=134), we determined that high PRL-receptor expression in the primary tumor was associated with a shorter time to bone metastasis (PRLR AQUA Max Hazard ratio=1.04, 95% Hazard Ratio confidence limits 1.00-1.07, p=0.03/multivariable Cox proportional hazards model), indicating their treatment failure may be related to the PRL-receptor. We also identified the PRL-receptor on rare samples of matched primary and bone metastases. In an analysis of advanced breast cancer patients, we also detected the PRL-receptor in circulating tumor cells of the blood. PRL treatment of breast cancer cells induced osteoclast differentiation and bone lysis via presumed secreted factors, and interestingly these effects were abrogated by a PRL-receptor-antagonist (delta1-9-G129R-hPRL). We identified sonic hedgehog as part of the molecular mechanism by which PRL and the PRL-receptor induce breast cancer cells to directly promote the differentiation of osteoclast cells capable of bone resorption. This molecular mechanism identifies key potential therapeutic targets to ameliorate the devastating effects of breast cancer to bone metastasis and potential predictive biomarkers.

Citation Format: Amanda Forsyth, Ashley Sutherland, Yingying Cong, Laurel Grant, Tzu-Hua Juan, Jae K. Lee, Alexander Klimowicz, Stephanie K. Petrillo, Jinghui Hu, Angela Chan, Florence Boutillon, Vincent Goffin, Cay Egan, Patricia A. Tang, Li Cai, Don Morris, Anthony Magliocco, Carrie S. Shemanko. Prolactin promotes breast cancer to bone metastasis and breast cancer cell-mediated osteoclast differentiation. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research; Oct 17-20, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(2_Suppl):Abstract nr B56.
Abstract B59: Age and estrogen-dependent inflammation in breast adenocarcinoma and normal breast tissue
Authors: Quigley David A. , Tahiri Andliena , Lüders Torben , Riis Margit H. , Balmain Allan , Børresen-Dale Anne-Lise , Bukholm Ida R. , Kristensen Vessela
Published: May 2016

Chronic inflammation promotes breast tumor growth and invasion by accelerating angiogenesis and tissue remodeling in the tumor microenvironment. There is a complex relationship between inflammation and estrogen, which drives the growth of 70 percent of breast tumors. Low levels of estrogen exposure stimulate macrophages and other inflammatory cell populations, but very high levels are immune suppressive. Breast tumor incidence is increased by obesity and age, which interact to influence inflammatory cell populations in normal breast tissue. To characterize the impact of these factors on tumors and the tumor microenvironment, we measured gene expression profiles in 195 breast adenocarcinomas and 195 matched adjacent normal breast tissue samples. Age and Body Mass Index (BMI) were independently associated with inflammation in normal tissue but not tumors. We developed a novel statistic, the Rewiring Coefficient, to quantify the rewiring of gene co-expression networks at the level of individual genes. Estrogen Receptor (ER)-negative tumors had elevated macrophage expression compared to matched normal tissue, but ER-positive tumors showed an unexpected decrease in macrophage expression. We found an inverse relationship between the increase in tumor estrogen pathway expression compared to adjacent normal tissue and that tumor's macrophage score and validated this finding in 126 matched breast tumor-normal pairs from the previously published METABRIC cohort. Our data support an immune suppressive effect of high doses of estrogen signaling in breast tumor microenvironment, suggesting that this effect contributes to the greater presence of prognostic and therapeutically relevant immune cells in ER-negative tumors.

Citation Format: David A. Quigley, Andliena Tahiri, Torben Lüders, Margit H. Riis, Allan Balmain, Anne-Lise Børresen-Dale, Ida R. Bukholm, Vessela Kristensen. Age and estrogen-dependent inflammation in breast adenocarcinoma and normal breast tissue. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research; Oct 17-20, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(2_Suppl):Abstract nr B59.
Abstract IA10: Sox10 regulates stem/progenitor and mesenchymal cell states in mammary epithelial cells
Authors: Dravis Christopher , Wahl Geoffrey M.
Published: May 2016

To discover mechanisms that mediate the initiation and progression of aggressive and stem-like breast cancers, we characterized signaling networks that are present in the mammary stem cells responsible for fetal and adult mammary development. These analyses identified a signaling axis between FGF signaling and the transcription factor, Sox10. Here we report that Sox10 is specifically expressed in mammary cells that exhibit the highest levels of stem/progenitor activity. This includes fetal and adult mammary cells in vivo and mammary organoids in vitro. Sox10 is functionally relevant, as its deletion reduces stem/progenitor competence, while its overexpression increases stem/progenitor activity. Intriguingly, we also discover that Sox10 overexpression elicits epithelial-to-mesenchymal transition (EMT) in mammary organoids in an FGF signaling-dependent manner. Further, modulation of Sox10 levels can induce sequential EMT, migratory, and clonogenic behaviors that strikingly resemble proposed mechanisms of metastasis. Consistent with these findings, we find that Sox10 is preferentially expressed in the most stem- and EMT-like triple negative breast cancer subtypes, and that Sox10+ tumor cells utilize FGF signaling for growth and invasive behaviors, which suggest that Sox10 may reprise these same functions during tumorigenesis. Collectively, these results demonstrate a signaling mechanism through which stem and mesenchymal-like states are acquired in mammary cells, and indicate possible therapeutic targets to counter these functions in breast cancers for which targeted therapies are currently unavailable.

Citation Format: Christopher Dravis, Geoffrey M. Wahl. Sox10 regulates stem/progenitor and mesenchymal cell states in mammary epithelial cells. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research; Oct 17-20, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(2_Suppl):Abstract nr IA10.
Abstract IA16: Clinical relevance of host immunity in breast cancer
Authors: Loi Sherene
Published: May 2016

The clinical relevance of the host immune system in breast cancer has long been unclear. Recently an appreciation of the biological heterogeneity of breast cancer has prompted investigation of the immune system's role in the different molecular breast cancer subtypes. A robust association between a more favourable prognosis and the presence of high levels of lymphocytic infiltration has been found in early-stage triple negative and HER2-positive breast cancer. These infiltrates seem to represent favorable anti-tumor immune responses, suggesting that activation of host immunity will be important for improving survival outcomes. The biological influences underlying the immune response are incompletely understood.My talk will review recent data supporting the clinical relevance of host anti-tumor immunity as represented by lymphocytic infiltration, the composition of the infiltrate in breast cancer and how this biomarker could be used in the clinical setting. Lymphocytic infiltration may be important as a biomarker in breast cancer prognosis: efforts to standardize the methodology for evaluating TILs in clinical breast cancer specimens in routine histopathological practise are ongoing. I will also discuss the rationale for enhancing immunity in breast cancer, including data on the efficacy of T-cell checkpoint inhibition..

References:

1. The evaluation of tumor-infiltrating lymphocytes (TILs) in breast cancer: recommendations by an International TILs Working Group 2014. Ann Oncol. 2015 Feb;26(2):259-71. doi: 10.1093/annonc/mdu450. Epub 2014 Sep 11.

2. Host antitumor immunity plays a role in the survival of patients with newly diagnosed triple-negative breast cancer. J Clin Oncol. 2014 Sep 20;32(27):2936-8.

3. Tumor-Infiltrating Lymphocytes and Associations With Pathological Complete Response and Event-Free Survival in HER2-Positive Early-Stage Breast Cancer Treated With Lapatinib and Trastuzumab: A Secondary Analysis of the NeoALTTO Trial. JAMA Oncol. 2015 Jul 1;1(4):448-54. doi: 10.1001/jamaoncol.2015.0830.

Citation Format: Sherene Loi. Clinical relevance of host immunity in breast cancer. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research; Oct 17-20, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(2_Suppl):Abstract nr IA16.
Abstract A33: A role of SET/I2PP2A in HNSCC angiogenesis and tumor-promoting microenvironment
Authors: Sobral Lays , Santos Leandro F. , Curti Carlos , Leopoldino Andreia M.
Published: May 2016

This study addressed a potential role of SET/I2PP2A protein on angiogenesis and immune response in head and neck squamous cell carcinoma (HNSCC). A HNSCC cell line with stable SET knockdown (HN12shSET/HN12shControl), previously established in our laboratory, was used to perform (1) qPCR array analysis of genes related to angiogenesis (Qiagen), and (2) tube-formation assay using Human Umbilical Vein Endothelial cells (HUVEC) cultured with HN12shSET cells conditioned medium. Moreover, several chemokines and cytokines were analyzed by Multi-Analyte ELISA array (Qiagen), and the specific levels of TGF-B1 and RANTES were validated by a Single-Analyte ELISA array (Qiagen) in cells exposed or not to TNF-alpha; stimulus. The cellular distribution of NF-kB, p-NF-kB and p-STAT3 was assessed by immunofluorescence, cell fractionation and western blotting assays. Our results showed higher levels of genes that induce angiogenesis, in particular VEGFC, in HN12shSET cells; these cells presented, in addition, higher TGF-B1 and lower RANTES/CCL-5 secretion levels compared to controls, as well as dowregulation of CCL-2 expression. Moreover, the stimulation of HUVEC tube-formation by the HN12shSET cells conditioned medium was higher (1.7 fold) compared to control, and p-NF-kB and p-STAT3 were found predominantly in the nucleus of HN12shSET cells; this suggests a negative balance between SET and regulation of these transcription factors. Therefore, SET knockdown in HN12 cells stimulates angiogenesis through VEGFC and TGF-B1 secretion, which could contribute to create a tumor-promoting microenvironment. These results support a SET role in HNSCC development and progression

Citation Format: Lays Sobral, Leandro F. Santos, Carlos Curti, Andreia M. Leopoldino. A role of SET/I2PP2A in HNSCC angiogenesis and tumor-promoting microenvironment. [abstract]. In: Proceedings of the AACR Special Conference: Developmental Biology and Cancer; Nov 30-Dec 3, 2015; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(4_Suppl):Abstract nr A33.
Abstract B12: March E3 ubiquitin ligase is required for head formation by mediating Dishevelled degradation during Xenopus development
Authors: Lee Hyeyoon , Cheong Seong-Moon , Han Jin-Kwan
Published: May 2016

Wnt signaling controls numerous biological events including vertebrate development, adult tissue homeostasis and organogenesis. In addition, dysregulation of Wnt itself or its downstream signaling components causes human birth defects and cancer. Dishevelled (Dvl) is a crucial scaffolding protein that exerts activation of Wnt signaling pathway. Recently, several Dvl-interacting proteins have been suggested to mediate the stability of Dvl. However, the molecular mechanisms by which Dvl undergoes degradation remain to be elucidated. Here we identify March, a RING domain-containing ubiquitin ligase, targeting Dvl protein and show that it regulates canonical Wnt signaling in Xenopus. During Xenopus embryogenesis, March transcripts were ubiquitously expressed. We intriguingly found that March depletion by microinjection of morpholino oligonucleotide caused defects in the anterior head formation, including loss of the forebrain, cement gland and eye structures. We also found that these defective phenotypes were caused by the Wnt antagonistic function of March in the head organizer region. Knockdown of March suppressed expression of head organizer genes Gsc, Chd and Otx2 at the dorsal marginal region of the gastrula embryo. Wnt reporter assay, western blot analysis measuring phosphorylation of LRP6 and activation of β-catenin, RT-PCR and axis duplication assay in Xenopus revealed that March inhibits canonical Wnt signaling pathway. Furthermore, we identified that March protein associates with Dvl and mediates its poly-ubiquitination for lysosomal degradation in Xenopus embryos. Taken together, these results suggest that March retains cytosolic pools of Dvl protein and leads subsequent limitation of Wnt signaling for vertebrate head development.

Citation Format: Hyeyoon Lee, Seong-Moon Cheong, Jin-Kwan Han. March E3 ubiquitin ligase is required for head formation by mediating Dishevelled degradation during Xenopus development. [abstract]. In: Proceedings of the AACR Special Conference: Developmental Biology and Cancer; Nov 30-Dec 3, 2015; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(4_Suppl):Abstract nr B12.
Abstract B26: Centriole amplification during mammalian multiciliated cell development reveals a novel centrosome asymmetry
Authors: Jord Adel Al , Spassky Nathalie , Meunier Alice
Published: May 2016

The centrosome comprises a mother and a daughter centriole, both regarded as equivalent in their ability to form new centrioles. Their symmetric duplication during the cell cycle is crucial for proper genetic partitioning between the daughter cells. In multiciliated cells, hundreds of centrioles are thought to appear de novo, i.e. independently from the centrosome, to nucleate the same number of motile cilia and propel physiological fluids. The origin of these centrioles remains unknown. Here we provide the cellular mechanism driving large-scale centriole production in multiciliated cells and identify the daughter centriole of the centrosome as the nucleation point for 95% of the new centrioles. Using live imaging combined with correlative super-resolution light and electron microscopy microscopy, we show that centriole amplification proceeds through a recurrent mechanism whereby the daughter centriole concomitantly generates the new centrioles and their cytoplasmic shuttles known as deuterosomes. By revealing this novel centrosome cycle, our work challenges the postulate that large-scale centriole production is a de novo mechanism in mammalian cells. In addition, it uncovers a fundamental asymmetry between mother and daughter centrioles in the control of centriole biogenesis and number. These results may have critical implications in the understanding of the etiology of cilia-related diseases, brain defects, and tumor formation.

Citation Format: Adel Al Jord, Nathalie Spassky, Alice Meunier. Centriole amplification during mammalian multiciliated cell development reveals a novel centrosome asymmetry. [abstract]. In: Proceedings of the AACR Special Conference: Developmental Biology and Cancer; Nov 30-Dec 3, 2015; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(4_Suppl):Abstract nr B26.
Abstract B29: Snail regulates extracellular matrix-mediated VEGFR3 expression in developmental and tumor angiogenesis
Authors: Park Jeong Ae , Lee Ho-Young , Kwon Young-Guen
Published: May 2016

Snail family is a zinc finger transcription factor and involved in epithelial branching morphogenesis and EMT by altering the ability of polarity, motility, and adhesion. The expression of Snail family in developing and tumor vessels has been suggested, however, the precise expression pattern and cellular function of Snail remain to be unclear. We identified Snail as an angiogenic regulator through inducing VEGFR3 under extracellular matrix (ECM)-meditated signalings. Snail was upregulated in the postnatally developing retinal endothelial cells (ECs) and in the vessels lining ductal breast tumors. In vitro endothelial sprouting assay indicated that knockdown or ectopic expression of Snail impaired EC sprouting on ECM. Moreover, Snail colocalized with VEGFR3 and upregulated VEGFR3 mRNA by cooperating with early growth response protein-1 in ECs. In the developing vessel model, intraperitoneal injection with stable Snail siRNA in postnatal mice demonstrated that Snail loss attenuated the formation of the vascular plexus not only by impairing vertical sprouting vessels but also by downregulating VEGFR3. Furthermore, in MMTV-PyMT breast tumors, Snail was strongly expressed in endothelial vessels around ductal breast tumors along with VEGFR3 expression. These data demonstrate that Snail contributes to vascular morphogenesis by upregulating VEGFR3. ECM-Snail-VEGFR3 axis may play crucial roles in disorganized vascular network in solid tumors.

Citation Format: Jeong Ae Park, Ho-Young Lee, Young-Guen Kwon. Snail regulates extracellular matrix-mediated VEGFR3 expression in developmental and tumor angiogenesis. [abstract]. In: Proceedings of the AACR Special Conference: Developmental Biology and Cancer; Nov 30-Dec 3, 2015; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(4_Suppl):Abstract nr B29.