bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract PD8-06: Fluorescence guided breast conserving surgery
Authors: Sharma CK , Ranjan P , Qaiser D , Dhar A , Srivastava A , Kataria K
Published: May 2019

Background. Palpation guided breast conserving surgery for breast cancer is associated with tumor involved margins in up to 41% of cases. Ultrasound guided breast conserving surgery and frozen section biopsy result in a significant reduction in margin positivity. However, intra-operative USG and Frozen section biopsy are not widely available. We aimed to find a simple and effective technique for intra-operative margin assessment during breast conserving surgery.

Hypothesis. Intravenously injected Fluorescein reaches tumor tissue in high concentration due to increased tumor blood flow and increased capillary permeability which can be detected by blue light (494 nm).

Objective. To find diagnostic accuracy of fluorescence guided identification of tumor margins during breast conserving surgery.

Methods. A total of 60 patients with T1–T2 invasive breast cancer who underwent breast conserving surgery at AIIMS, New Delhi between March, 2016 to Feb, 2018 were included. Each patient received 2 ml of intra-venous 20% Fluorescein sodium just before skin incision. Breast conserving surgery was performed under USG guidance. Specimen was bisected and examined under blue light. Fluorescent tumor margins were identified and six biopsies taken from non-fluorescent area 5mm outside from fluorescent tumor margins at 2,4,6,8,10 and 12 o'clock position. The involvement of USG guided margin was compared with Fluorescence guided margins on histopathology.

Results. Mean age of the patient was 51.2 (SD=6.4) years. Fourteen patients (20.3 %) had T1 tumors while 46 patients(69.7%) had T2 tumors. Axillary nodes were involved in 19 patients (20.1%) and all underwent axillary node dissection. Eight patients (13.3%) received NACT. Fluorescent margins in two (3.3%) out of 60 patients were involved by tumor while USG guided margins in all the 60 patients were free. The specificity of fluorescence in identifying uninvolved tumor margins during breast conserving surgery is 96.7%.

No patient developed adverse drug reactions.

Conclusion. The fluorescence identifies tumor free margins with 96.7% accuracy. It is simple, effective and affordable method of identifying margin positivity in breast conservation surgery.

Citation Format: Sharma CK, Ranjan P, Qaiser D, Dhar A, Srivastava A, Kataria K. Fluorescence guided breast conserving surgery [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr PD8-06.
Abstract PL1: Breast Tumor Evolution and Intratumor Heterogeneity - Insights from Single Cell Genomics
Authors: Navin N
Published: May 2019

Breast tumors evolve from single cells in the ducts and acquire mutations leading to divergent lineages and intratumor heterogeneity in invasive cancers. To understand the genomic evolution of breast cancers during complex evolutionary processes including initiation, invasion and therapeutic resistance we have developed single cell sequencing (SCS) methods. I will discuss our efforts in applying SCS methods to study invasion in 10 premalignant breast cancer patients with ductal-carcinoma-in-situ (DCIS), which led to a discovery of early punctuated copy number evolution in the ducts and multi-clonal invasion. I will also discuss our work in applying both single cell DNA and RNA sequencing methods to study chemoresistance evolution in 20 triple-negative breast cancer patients using longitudinal samples, which identified an adaptive genomic evolution and acquired transcriptional reprogramming during resistance. Finally I will present unpublished work on our efforts to develop a 'Human Breast Cell Atlas' of cell types and cell states (transcriptional programs) of normal cell types, and recent work on delineating copy number evolution in BRCA-positive breast cancer patients.

Citation Format: Navin N. Breast Tumor Evolution and Intratumor Heterogeneity - Insights from Single Cell Genomics [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr PL1.
Abstract A08: CCL2-induced chemokine cascade promotes breast cancer metastasis via retention of metastasis-associated macrophages
Authors: Kitamura Takanori , Pollard Jeffrey W.
Published: May 2019

This abstract is being presented as a short talk in the scientific program. A full abstract is printed in the Proffered Abstracts section (PR17) of the Conference Proceedings.

Citation Format: Takanori Kitamura, Jeffrey W. Pollard. CCL2-induced chemokine cascade promotes breast cancer metastasis via retention of metastasis-associated macrophages. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr A08.
Abstract A14: A cholesterol metabolite acts on the host to increase breast cancer metastasis
Authors: Baek Amy Eun Joo , Nelson Erik Russell
Published: May 2019

With a woman's lifetime risk of developing invasive breast cancer at one in eight, breast cancer remains the most common cancer and the second leading cause of cancer death in women. Despite the success of frontline endocrine therapies, relapse with resistant metastatic disease is common, and triple negative disease remains without a targeted therapy. Importantly, breast cancer associated mortality is most often due to the metastatic spread of the disease. Therefore, there continues to be a need for studies that may lead to the development of new chemopreventive strategies and/or lifestyle changes that reduce breast cancer metastasis. In this regard, both obesity and elevated cholesterol are risk factors for metastatic relapse, while patients receiving cholesterol-lowering drugs such as HMG-CoA reductase inhibitors (statins) display an increased relapse-free survival time. Therefore, clinical observations strongly implicate cholesterol in the metastatic progression of, although the underlying mechanisms remain unknown. Thus, we initiated a series of studies to evaluate the mechanisms by which cholesterol promotes breast cancer metastasis. Using murine models of mammary cancer, we find that elevated cholesterol does indeed increase metastasis. Further, statins attenuated the metastatic effects of a high fat diet, implicating cholesterol as a primary mediator of this pathology. Importantly, the pro-metastatic effects of cholesterol required the presence of CYP27A1, the enzyme responsible for its conversion to 27-hydroxycholesterol (27HC). We have recently shown that 27HC is an endogenous ligand for the both the estrogen receptors (ERs) and liver X receptors (LXRs). Interestingly, however, the metastatic effects of 27HC were also prevalent in ER-negative models. While LXR activation did have a modest effect on epithelial to mesenchymal transition (EMT), we find that the robust pro-metastatic effects of 27HC are mediated in a cancer cell-extrinsic manner. Specifically, we find that 27HC alters myeloid cell recruitment and function at distal metastatic sites, and thereby facilitates cancer cell colonization and growth by modulating the host environment. In summary, our data strongly suggest that by altering the host immune system, 27HC is the down-stream mediator of the effects of obesity and cholesterol on breast cancer metastasis. These results provide additional support for development of specific CYP27A1 inhibitors and immune-modulatory therapies for the prevention and treatment of breast cancer metastasis.

This work was supported by an NIH-NCI grant to ERN (4R00CA172357-03).

Citation Format: Amy Eun Joo Baek, Erik Russell Nelson. A cholesterol metabolite acts on the host to increase breast cancer metastasis. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr A14.
Abstract A19: Pulmonary microthrombosis enhances tumorigenesis via myeloid hypoxia-inducible factors
Authors: Evans Colin E. , Palazon Asis , Sim Jingwei , Tyrakis Petros A. , Bendahl Par-Ola , Belting Mattias , Rundqvist Helene , Branco Cristina , Johnson Randall S.
Published: May 2019

Thrombosis causes local blood flow restriction and tissue hypoxia, and both of these are associated with tumor cell metastasis. To better understand the regulation of thrombosis-induced metastasis, we created a model incorporating elements of both processes. Controlled induction of pulmonary microthrombosis caused an increase in expression of first hypoxia-inducible factor (HIF)1α, and subsequently HIF2α. Induction of thrombosis before the introduction of tumor cells to venous circulation had no effect on pulmonary tumor number or size; but thrombosis at the time of tumor cell seeding increased number and size of tumors in the lung. Thrombosis on the day after seeding of tumor cells caused an even greater increase in tumor number and size, and this effect persisted until even when thrombosis was induced five days after the introduction of tumor cells into the blood. Experiments on myeloid HIF1α or HIF2α knockout mice demonstrated that loss of either HIF1α or HIF2α eliminated the advantage given to pulmonary tumorigenesis by thrombotic insult. In primary human tumours, markers of thrombosis were positively correlated with expression of the 2 HIFα isoforms. These data demonstrate the importance of microthrombosis in a novel model of metastasis and the essential role of the myeloid cell-specific HIFα response in mediating this process.

Citation Format: Colin E. Evans, Asis Palazon, Jingwei Sim, Petros A. Tyrakis, Par-Ola Bendahl, Mattias Belting, Helene Rundqvist, Cristina Branco, Randall S. Johnson. Pulmonary microthrombosis enhances tumorigenesis via myeloid hypoxia-inducible factors. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr A19.
Abstract A20: Mammary tumor-derived CCL2 enhances pro-metastatic systemic inflammation through upregulation of macrophage-derived IL1beta
Authors: Kersten Kelly , Coffelt Seth B. , Verstegen Niels J. , Vrijland Kim , Ciampricotti Metamia , Doornebal Chris W. , Hau Cheei-Sing , Doshi Parul , Visser Karin E. de
Published: May 2019

Metastatic breast cancer remains a major cause of cancer-related death among women. Metastasis is regulated by extensive crosstalk between cancer cells and immune cells. Besides the onset of a local inflammatory microenvironment, tumors frequently induce a systemic inflammatory state characterized by the release of various cytokines, chemokines and growth factors to mobilize myeloid cells that support metastasis, emphasizing the notion that cancer should be regarded as a systemic disease. Utilizing the K14cre;Cdh1F/F;Trp53F/F (KEP) conditional mouse model of metastatic breast cancer, we have recently unraveled a novel mechanistic link between mammary tumor-induced IL17-producing gamma delta T cells, systemic expansion of immunosuppressive neutrophils and metastasis formation. We identified tumor-derived IL1beta as one of the drivers of this cascade by inducing IL17 release from gamma delta T cells. However, it remains unclear how other inflammatory mediators are involved in this systemic inflammatory cascade. In the current study, we identified the pro-inflammatory cytokine CCL2 as a key regulator of the gamma delta T cell IL17 neutrophil axis. CCL2 blockade resulted in reduced IL17-production by gamma delta T cells, decreased systemic neutrophil accumulation and enhanced CD8+ T cell activity. We show that activation of this cascade relies on CCL2-mediated release of IL1beta from CCR2+ tumor-associated macrophages. These results indicate that CCL2, via IL1beta, acts as a key player in regulating the pro-metastatic gamma delta T cell IL17 neutrophil systemic inflammatory cascade and might be an interesting candidate for therapeutic targeting.

Citation Format: Kelly Kersten, Seth B. Coffelt, Niels J. Verstegen, Kim Vrijland, Metamia Ciampricotti, Chris W. Doornebal, Cheei-Sing Hau, Parul Doshi, Karin E. de Visser. Mammary tumor-derived CCL2 enhances pro-metastatic systemic inflammation through upregulation of macrophage-derived IL1beta. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr A20.
Abstract A22: The effects of Lipocalin-2 in the early stages of colorectal cancer metastasis
Authors: Hughes Daniel , Davis Celestia , Zhang Yu , Bonaparte John , Pena Maria
Published: May 2019

Colorectal cancer (CRC) is the second leading cause of cancer deaths in the United States. When diagnosed at an early, localized stage, the five year survival rate is approximately 90%; however, when patients are diagnosed at the metastatic stage, this drops to 12%. Metastasis, the spread of cancer cells from the primary tumor to a distant organ, is the primary cause of death in cancer patients. In CRC, metastasis is typically found in the liver, asymptomatic, and often detected at the final stage when therapeutic options are few. The genetic and cellular mechanisms driving the early stages of metastasis are still poorly understood. Recent studies have shown that prior to the arrival of cancer cells at the secondary organ, molecular signals from the tumor direct the recruitment of bone marrow derived cells to create a pre-metastatic niche where cancer cells can attach and develop into a metastatic lesion. Identifying and understanding these molecules can lead to the development of methods for early diagnosis or targets for intervention.

Using an orthotopic model of CRC liver metastasis, we performed microarray analyses of the liver microenvironment in tumor bearing mice before and after the arrival of metastatic cells. We found that Lipocalin-2 (LCN2, 24p3, NGAL) was highly expressed in the liver and in circulation of mice bearing highly metastatic cells. LCN2 is a 25 kD secreted glycoprotein that binds small lipophilic molecules. Its expression is upregulated in a number of primary tumors of epithelial origin, but its role in metastasis remains unclear. It has contrasting roles in metastasis in breast cancer models and has been associated with poor prognosis in CRC patients; however its role in the tumor microenvironment has not been studied. Our studies thus far show that overexpression of Lcn2 in mouse colon cancer cells either had little impact on tumor growth and invasiveness, or it enhanced invasion in the presence of macrophages; however, it resulted in an overall reduction of circulating iron levels in sera from tumor bearing mice. In-vitro, iron also seems to have an effect in conjunction with Lcn2 on the invasiveness of the tumor cells. The results from this study will allow us to better describe the role of LCN2 in tumor and stromal cells in the early stages of liver metastasis.

Citation Format: Daniel Hughes, Celestia Davis, Yu Zhang, John Bonaparte, Maria Pena. The effects of Lipocalin-2 in the early stages of colorectal cancer metastasis. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr A22.
Abstract A32: Targeting c-Met by Pterostilbene (PTER) suppresses breast cancer brain metastasis
Authors: Xing Fei , Sharma Sambad , Liu Yin , Wu Kerui , Watabe Kounosuke
Published: May 2019

At the late stage of breast cancer, most patients develop metastatic lesions and the brain is one of the major metastatic sites. Brain metastasis profoundly affects the cognitive and sensory function as well as morbidity of patients, and the one year survival rate among these patients remains less than 20%. The result of our newly developed gene set enrichment analysis (GSEA)-based pathway screening indicates that c-Met pathway is highly activated among the patients who developed early brain metastasis. To identify a novel target of c-Met pathway in brain metastasis, we performed the microarray analysis in two brain metastatic cell lines with the expression of doxycycline-inducible sh-cMet. Our results indicate that a group of inflammatory cytokines including IL1β, IL8 and CXCL1 were significantly induced by c-Met activation. Furthermore, we found that IL1β was able to induce the secretion of HGF (hepatocyte growth factor) from tumor associated astrocytes (TAA) which in turn activates c-Met pathway in cancer cells. Our results of endothelial cell tube formation assays also strongly suggest that c-Met-induced IL8 and CXCL1, promote tumor angiogenesis which is essential for the metastatic growth of cancer cells. Natural compounds (NC) have been extensively studied for their anti-tumor effects. However, much less studies have been done on NC for the treatment/prevention of brain tumors mainly due to the obstacle of Blood-brain-barrier (BBB) permeability. To identify the NCs that can be used for treating brain metastasis, we performed pathway analysis which only focused on BBB-permeable NC. The results of GSEA indicate that Resveratrol-targeting genes were significantly enriched among those patients who developed brain metastasis compared to metastasis-free patients. Furthermore, we found that the one of derivatives of Resveratrol, Pterostilbene (PTER), showed significantly more potent activity than Resveratrol and suppressed brain metastasis in vitro and in vivo by targeting the c-Met oncogene. These findings suggest that TAA-mediated c-Met activation plays a key role in brain metastasis and that PTER is a potential therapeutic agent to treat brain metastasis by suppressing the c-Met expression.

Citation Format: Fei Xing, Sambad Sharma, Yin Liu, Kerui Wu, Kounosuke Watabe. Targeting c-Met by Pterostilbene (PTER) suppresses breast cancer brain metastasis. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr A32.
Abstract A35: Axl overexpression in a prostate cancer mouse model delays metastasis and may prolong dormancy of disseminated tumor cells
Authors: Axelrod Haley D. , Valkenburg Kenneth C. , Pienta Kenneth J.
Published: May 2019

The most common site of metastasis in prostate cancer (PCa) patients is the bone marrow, where disseminated tumor cells (DTCs) preferentially home to the hematopoietic stem cell niche and can undergo a state of dormancy. These PCa cells may lie dormant in the bone marrow for years after prostatectomy, but may eventually re-enter the cell cycle resulting in lethal metastatic disease. Using mouse models of PCa metastasis, our group has previously shown that Gas6/Axl signaling is associated with dormant bone marrow DTCs. In this study we investigated the requirement of Axl in initiating and maintaining the dormant phase. We hypothesized that overexpression of Axl in PCa cells would prolong dormancy. Overexpression of Axl in the bone-homing AR-positive Axl-negative human PCa cell line C42B slows proliferation in vitro. Subcutaneous injection of C42B-Axl cells in a xenograft mouse model resulted in delayed onset of tumor growth relative to WT controls. C42B-WT tumors reached an average volume of 200mm3 at 6.6 weeks, while C42B-Axl tumors reached the same volume at 11 weeks. Regions of the primary tumor with a high percentage of Axl+ cells correlated with a low number of Ki67+ cells, indicating that Axl may inhibit proliferation even in the primary tumor. Surprisingly, C42B-Axl tumors were more vascular as observed by eye, and regions of increased vasculature correlated with a high percentage of Axl+ cancer cells. Metastatic outgrowth in soft tissues reached a lethal burden ~12 weeks following C42B-WT tumor removal, while mice bearing C42B-Axl tumors did not have detectable metastases by eye at the same timepoint post-tumor removal and were apparently healthy. Metastatic outgrowth in the bone was detected by the presence of osteoblastic and osteolytic activity at 5-12 weeks post-tumor removal in the femurs and tibiae of mice bearing C42B-WT tumors. At 8-12 weeks after C42B-Axl tumor removal, however, there was no evidence of metastatic bone disease. While these experiments are still ongoing, we have shown that Axl expression in the primary tumor delayed metastatic burden in both soft tissues and in bone in a mouse model of PCa metastasis. In ongoing studies, we will compare DTC number and time to metastasis in the bone marrow of C42B-WT and C42B-Axl tumor-bearing mice.

Citation Format: Haley D. Axelrod, Kenneth C. Valkenburg, Kenneth J. Pienta. Axl overexpression in a prostate cancer mouse model delays metastasis and may prolong dormancy of disseminated tumor cells. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr A35.
Abstract A42: The role of M1 and M2 macrophage in the cycling phase of breast cancer
Authors: Walker Nykia D. , Elias Michael , Greco Steve , Rameshwar Pranela
Published: May 2019

Breast cancer malignancy is the second leading cause of death among women. The majority of deaths associated with cancer, from solid tumors are the result of metastasis and not from the primary tumor site Clinical evidence indicates that breast cancer (BC) cells (BCC) can exist in dormancy within the bone marrow (BM), for >10 years. Methods by which BCCs adapt dormancy as well as reversed dormancy into metastatic cells are highly significant for the development of new and safe therapies. This research focuses on the BM because it is the preferred organ for dormant BCCs. Furthermore, in several cases, the BM has been identified as a source of BCCs during clinical resurgence. A subset of BCCs with tumor initiating property (cancer stem cells, CSCs) has been identified as those responsible for dormancy and chemoresistance. We will take advantage of our working hierarchy of BCCs to determine how M2 macrophage (MΦ) facilitates dormancy or clinical resurgence into metastatic cells. The mechanisms by which dormancy occurs followed by later resurgence into metastatic role for the BM microenvironment to facilitate CSC dormancy. CSCs can form gap junctional intercellular communication (GJIC) with stroma, by release of miRNA, which induces cycling quiescence. MΦ are classified into two subsets, either M1 or M2 type. M1 are mostly pro-inflammatory (classically activated to kill microorganisms and tumor cells), while M2 are non-inflammatory (alternatively activated to promote tissue repair). M2 MΦ is one cell type that comprises the stromal compartment inside the BM hematopoietic microenvironment. We studied how activated MSCs can influence M1 or M2 polarization. Our supporting studies indicate that M2 MΦ can arrest BCCs in G1/G0, while M1 MΦ can induce cell cycle progression. Activated MSCs transitioned M2 into M1 phenotype whereas inactivated MSCs maintains M2 MΦ phenotype. Here we present how the transitional changes in M2 to M1 MΦ affect the behavior of the CSCs within the BM stromal compartment.

Citation Format: Nykia D. Walker, Michael Elias, Steve Greco, Pranela Rameshwar. The role of M1 and M2 macrophage in the cycling phase of breast cancer. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr A42.
Abstract A44: Metastatic latency and immune evasion mediated by autocrine WNT inhibition
Authors: Malladi Srinivas , Macalinao Danilo G. , Jin Xin , He Lan , Massague Joan
Published: May 2019

This abstract is being presented as a short talk in the scientific program. A full abstract is printed in the Proffered Abstracts section (PR06) of the Conference Proceedings.

Citation Format: Srinivas Malladi, Danilo G. Macalinao, Xin Jin, Lan He, Joan Massague. Metastatic latency and immune evasion mediated by autocrine WNT inhibition. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr A44.
Abstract A52: Dissection of cancer cells extravasation through human vascularized 3D microfluidic model: The major role of talin-1
Authors: Gilardi Mara , Bersini Simone , Kamm Roger Dale , Moretti Matteo , Vanoni Marco
Published: May 2019

This abstract is being presented as a short talk in the scientific program. A full abstract is printed in the Proffered Abstracts section (PR07) of the Conference Proceedings.

Citation Format: Mara Gilardi, Simone Bersini, Roger Dale Kamm, Matteo Moretti, Marco Vanoni. Dissection of cancer cells extravasation through human vascularized 3D microfluidic model: The major role of talin-1. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr A52.
Abstract A54: Intravital microscopy of prostate cancer lesions in bone: Kinetics of osteolysis and therapy response
Authors: Dondossola Eleonora , Alexander Stephanie , Alexander Steve , Holzapfel Boris , Logothetis Christopher , Hutmacher Dietmar , Friedl Peter
Published: May 2019

This abstract is being presented as a short talk in the scientific program. A full abstract is printed in the Proffered Abstracts section (PR04) of the Conference Proceedings.

Citation Format: Eleonora Dondossola, Stephanie Alexander, Steve Alexander, Boris Holzapfel, Christopher Logothetis, Dietmar Hutmacher, Peter Friedl. Intravital microscopy of prostate cancer lesions in bone: Kinetics of osteolysis and therapy response. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr A54.
Abstract A56: Identification of an ErbB2+ early disseminating cancer cell subpopulation with metastatic potential
Authors: Harper Kathryn , Sosa Maria Soledad , Hosseini Hedayatollah , Valderas Alvaro Avivar , Nagi Chandandaneep , Davis Roger , Klein Christoph , Entenberg David , Condeelis John , Farias Eduardo , Aguirre-Ghiso Julio
Published: May 2019

Recent clinical evidence suggests that cells are capable of disseminating from pre-invasive breast cancer lesions. It has been proposed that these early disseminating cancer cells (eDCCs) lodge into secondary sites, enter a state of dormancy, escape conventional therapy, and potentially evolve through different pathways from the primary tumor to form metastases. However, the mechanisms that endow these early cancer cells to disseminate and their behavior in target organs remain unknown. Here we show that ErbB2+ early cancer cells, while non-tumorigenic are highly efficient in disseminating. While a majority of these eDCCs remain dormant in target organs they are still endowed with a metastatic capacity. We identified an ErbB2HIGH/p38LOW/E-cadherinLOW/PRHI population of early cancer cells, which undergo a Wnt-dependent EMT and disseminate. Based on this signature we propose that these early cancer cells tap into motility programs similar to what is seen during branching morphogenesis (PRHI/WntHI/p38LOW) in order to disseminate. Using intra-vital imaging of transgenic ErbB2-T-CFP pre-malignant lesions we imaged this process at high resolution revealing that ErbB2HIGH/p38LOW/E-cadherinLOW cells display activate cancer cell motility, invadopodia formation and intravasation capacity. This correlated with CK8/18+/HER2+ circulating cancer cells as well as eDCCs in the lung and bone marrow of mice, a process that was enhanced following 2 weeks of systemic p38α/β inhibition. We conclude that during stages previously considered non-invasive, some HER2+ cancer cells can aberrantly activate an invasive morphogenetic program causing early dissemination that is normally restricted by active p38 signaling. These eDCCs carry latent metastatic initiating capacity and may contribute to relapse, altering our understanding of metastasis onset and how it might be targeted effectively.

Citation Format: Kathryn Harper, Maria Soledad Sosa, Hedayatollah Hosseini, Alvaro Avivar Valderas, Chandandaneep Nagi, Roger Davis, Christoph Klein, David Entenberg, John Condeelis, Eduardo Farias, Julio Aguirre-Ghiso. Identification of an ErbB2+ early disseminating cancer cell subpopulation with metastatic potential. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr A56.
Abstract A64: Development of a cloud-based histology database for collaborative cancer research
Authors: Cheng Ke
Published: May 2019

Background: Cancer research is highly dependent on histopathology analysis for diagnosis, prognosis, and treatment. However, the majority of valuable histology data is stored in private archives on glass slides and local hard drives, and not shared like bioinformatics data. Lack of centrally-located data leads to repetitive, redundant research and makes it difficult for cancer researchers to work collaboratively by limiting widespread access and image sharing. Development of a virtual slide database on a scalable IT infrastructure will improve cancer histology data viewing and searching resulting in enhanced global scientific collaboration, allowing cancer to be fought cooperatively instead of individually.

Design: The main repository for WizBase is a relational database powered by Amazon Web Services with online viewing, collaboration and long-term archiving of WSIs. We will also create ontology parameters and tagging approach using 3 types metadata: Type I, classification parameters, specimen information and experimental details captured at order submission; Type II, ontology terms and annotation added upon visualization of the slide by HistoWiz and slide owners; and Type III, crowd-sourced tags and collaborative comments not requiring formality or standardization.

Results: We have developed a robust IT infrastructure allowing for viewing, tagging and searching of histology images. HistoWiz viewer is the first Cloud based viewer that allows users to instantly view their histology slides on any mobile device without the need to download any files, software or plugins. Furthermore, we have establish ontology parameters (such as metastasis, necrosis, neovascularization) for the classification and tagging of digital pathology cancer images, and slide metadata are captured during online order submission. Finally, through defined field and free text tagging of the slide metadata, users can search for slides meeting specific criteria. Searches based on similarity to a particular subject slide metadata allows ranking slide relevancy. The crowdsourced database currently has >10,000 histology slides and is growing at a rate of 300% per year.

Conclusion: WizBaseTM is the first centralized Cloudbased WSI database for cancer histopathology with an image tagging web application to facilitate histology-driven data mining. WizBaseTM can be regarded as the scientific hybrid of a microscopy product with the viewing power of Google Earth combined with the search and crowdsourcing of Flickr.

Citation Format: Ke Cheng. Development of a cloud-based histology database for collaborative cancer research. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr A64.
Abstract B03: Formation of lymph node metastases is not angiogenesis dependent
Authors: Jones Dennis , Jeong Han-Sin , Liao Shan , Wattson Daniel A. , Cui Cheryl H. , Duda Dan G. , Willett Christopher G. , Jain Rakesh K. , Padera Timothy P.
Published: May 2019

Background: To date, antiangiogenic therapy has failed to improve overall survival in cancer patients when used in the adjuvant setting (local-regional disease with no detectable systemic metastasis). The presence of lymph node metastases worsens prognosis, however their reliance on angiogenesis for growth has not been reported.

Methods: Here, we introduce a novel chronic lymph node window (CLNW) model to facilitate new discoveries in the growth and spread of lymph node metastases. We use the CLNW in multiple models of spontaneous lymphatic metastases in mice to study the vasculature of metastatic lymph nodes. We further test our results in patient samples. Finally, we test the ability of antiangiogenic therapy to inhibit metastatic growth in the CLNW.

Results: We reveal the surprising lack of sprouting angiogenesis during metastatic growth, despite the presence of hypoxia in some lesions. Treatment with two different antiangiogenic therapies showed no effect on the growth or vascular density of lymph node metastases. We confirmed these findings in clinical specimens, including the lack of reduction in blood vessel density in lymph node metastases in patients treated with bevacizumab.

Conclusion: We provide pre-clinical and clinical evidence that sprouting angiogenesis does not occur during the growth of lymph node metastases, and thus reveal a new mechanism of treatment resistance to antiangiogenic therapy in adjuvant settings. The targets of clinically approved angiogenesis inhibitors are not active during early cancer progression in the lymph node, suggesting that inhibitors of sprouting angiogenesis as a class will not be effective in treating lymph node metastases.

Citation Format: Dennis Jones, Han-Sin Jeong, Shan Liao, Daniel A. Wattson, Cheryl H. Cui, Dan G. Duda, Christopher G. Willett, Rakesh K. Jain, Timothy P. Padera. Formation of lymph node metastases is not angiogenesis dependent. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr B03.
Abstract B08: Keap1–Nrf2 interaction suppresses cell motility in lung adenocarcinomas by targeting the S100P protein
Authors: Hua Kuo-Tai , Kuo Min-Liang
Published: May 2019

Purpose: Kelch-like ECH-associated protein 1 (Keap1) is an E3 ligase participated in the cellular defense response against oxidative stress through nuclear factor erythroid-2–related factor 2 (Nrf2). However, the role of Keap1 in regulating cancer motility is still controversial. We investigated the contribution of the Keap1–Nrf2 axis in the progression of non-small cell lung cancer (NSCLC).

Experimental Design: The expression of Keap1 and Nrf2 was examined via immunohistochemistry, real-time PCR, and Western blot analysis in a cohort of NSCLC tissues and cells. A series of in vivo and in vitro assays was performed to elucidate the contribution of the Keap1–Nrf2 axis in lung cancer mobility and progression.

Results: Keap1 expression was decreased in specimens from NSCLC patients with lymph node metastasis compared with patients without metastasis. Higher Keap1 expression levels were correlated with the survival ofNSCLC patients. Moreover,manipulation of Keap1 expression affected cell migration/invasion abilities. Depletion of Nrf2 relieved the migration promotion imposed by Keap1 suppression. Mechanistic investigations found that S100Pwas downregulated in both Keap1-overexpressing and Nrf2-knockdown NSCLC cells. Overexpression of Keap1 and knockdown of Nrf2 both suppressed S100P expression in NSCLC cells. Knockdown of S100P inhibited cell migration in highly invasive NSCLC cells and also relieved the migration promotion imposed by Keap1 suppression in weakly invasive NSCLC cells.

Conclusions: Our findings suggest that Keap1 functions as a suppressor of tumor metastasis by targeting the Nrf2/S100P pathway in NSCLC cells. In addition, overexpression of Keap1 may be a novel NSCLC treatment strategy and/or useful biomarker for predicting NSCLC progression.

Citation Format: Kuo-Tai Hua, Min-Liang Kuo. Keap1–Nrf2 interaction suppresses cell motility in lung adenocarcinomas by targeting the S100P protein. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr B08.
Abstract B10: Transcriptome analysis of mouse PDA subtypes
Authors: Balli David , Aiello Nicole , Norgard Robert , Li Jingyang , Sahmoud Amine , Stanger Ben
Published: May 2019

Patients with Pancreatic Adenocarcinoma (PDA) often present with advanced stage disease including widespread metastasis and have dismal 5-year survival rates. The molecular mechanism promoting PDA progression and metastasis remain poorly understood. We have previously demonstrated that tumor cells undergo Epithelial-to-Mesenchymal Transition (EMT) in mouse models of PDA [Rhim et al., 2012]. EMT is a developmental process where cells loss epithelial characteristics and transdifferentiate into a mesenchymal-like state that is highly associated with increased invasion and metastasis. To better understand molecular mechanisms of PDA development and metastatic disease, we subjected tumors cells from a lineage labeled mouse model of PDA to RNA sequencing to identify global transcriptional changes. Interestingly, we discovered that there are at least two distinct subtypes of PDA based on transcriptome profiling. Comparison of gene expression profiles from mouse PDA subtypes to established subtypes from human PDA datasets [Collisson et al., 2011] revealed that mouse PDA is transcriptionally similar to the human disease. Differences in transcriptional profiling between PDA subtypes may dictate response to chemotherapeutics and metastatic competency.

Citation Format: David Balli, Nicole Aiello, Robert Norgard, Jingyang Li, Amine Sahmoud, Ben Stanger. Transcriptome analysis of mouse PDA subtypes. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr B10.
Abstract B12: Polymorphisms in the Arntl2 promoter affect metastatic susceptibility in breast cancer
Authors: Ha Ngoc-Han , Hunter Kent W.
Published: May 2019

Metastasis accounts for the majority of cancer-related deaths. Although exact mechanisms are unknown, studies in our laboratory have demonstrated that germline polymorphisms could alter tumorigenesis and metastasis. Our lab developed a mouse genetics approach to investigate the effect of inherited single nucleotide polymorphisms (SNPs) on metastasis by crossing inbred mice of various backgrounds and subsequent mapping of quantitative trait loci (QTL) responsible for the phenotype. Using this approach, we crossed a highly metastatic (FVB) and low metastatic (MOLF) mouse strain and identified Arntl2, a circadian rhythm transcription factor, as a gene with differential expression that predicted distant metastasis-free survival in breast cancer patients. shRNA-mediated reduction in Arntl2 expression suppressed lung metastases, establishing Arntl2 as a metastasis modifier. Initial analysis revealed no polymorphisms in coding regions that could account for the differential expression in FVB and MOLF. Therefore, we performed whole genome sequencing of MOLF to identify SNP differences between MOLF and FVB. Overlapping this analysis with DNAse hypersensitivity sites in mouse mammary tumor cell lines revealed 13 polymorphisms in the predicted promoter region of Arntl2. FVB and MOLF promoters exhibited differential promoter activity in vitro, suggesting that these SNPs could explain the difference in Arntl2 expression between these two strains. In order to validate the causative role of the SNPs in vivo, metastatic cell lines were engineered using CRISPR technology to specifically replace the FVB with the MOLF promoter. Substitution of this promoter reduced Arntl2 transcript levels and subsequently decreased pulmonary metastases, as expected. Since Arntl2 is a transcription factor, current studies are focused on identifying Arntl2-regulated genes to investigate the pathways that are involved in modulating metastasis. This study will have important implications regarding the function of circadian rhythm in cancer progression and provide a potential molecular mechanism to explain the increased risk of breast cancers in nightshift workers. Furthermore, to the best of our knowledge this would be the first example of genome editing to validate polymorphisms in transcriptional control elements associated with inherited susceptibility for metastasis.

Citation Format: Ngoc-Han Ha, Kent W. Hunter. Polymorphisms in the Arntl2 promoter affect metastatic susceptibility in breast cancer. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr B12.
Abstract B15: RSK2 signals through stathmin to promote microtubule dynamics and tumor metastasis
Authors: Alesi Gina , Jin Lingtao , Li Dan , Magliocca Kelly , Kang Yibin , Chen Zhuo G. , Shin Dong M. , Khuri Fadlo R. , Kang Sumin
Published: May 2019

Metastasis is responsible for over 90% of cancer related deaths. Complex signaling in cancer cells orchestrates the progression from a primary to a metastatic cancer. However, the mechanisms of these cellular changes remain elusive. We previously demonstrated that p90 ribosomal S6 kinase 2 (RSK2) promotes tumor metastasis. Here, we investigated the role of RSK2 in the regulation of microtubule dynamics and its potential implication in cancer cell invasion and tumor metastasis. Stable knockdown of RSK2 disrupted microtubule stability and decreased phosphorylation of stathmin, a microtubule destabilizing protein, at serine 16 in metastatic human cancer cells. We found that RSK2 directly binds and phosphorylates stathmin at the leading edge of cancer cells. Phosphorylation of stathmin by RSK2 reduced stathmin-mediated microtubule depolymerization. Moreover, overexpression of phospho-mimetic mutant stathmin S16D significantly rescued the decreased invasive and metastatic potential mediated by RSK2 knockdown in vitro and in vivo. Furthermore, stathmin phosphorylation positively correlated with RSK2 expression and metastatic cancer progression in primary patient tumor samples. Our finding demonstrates that RSK2 directly phosphorylates stathmin and regulates microtubule polymerization to provide a pro-invasive and pro-metastatic advantage to cancer cells. Therefore, the RSK2-stathmin pathway represents a promising therapeutic target and a prognostic marker for metastatic human cancers.

Citation Format: Gina Alesi, Lingtao Jin, Dan Li, Kelly Magliocca, Yibin Kang, Zhuo G. Chen, Dong M. Shin, Fadlo R. Khuri, Sumin Kang. RSK2 signals through stathmin to promote microtubule dynamics and tumor metastasis. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr B15.
Abstract B23: Elucidating metastatic signaling networks in TNBC by investigating RKIP-regulated kinome
Authors: Yesilkanal Ali E. , Frankenberger Casey , Rabe Daniel C. , Johnson Gary L. , Rosner Marsha R.
Published: May 2019

Metastatic progression of tumors is the major cause of death in patients with triple negative breast cancer (TNBC), the most aggressive subtype of breast cancer. However, since metastasis is a multi-step process, unraveling its complexity is a major challenge. One effective way of tackling this question is to study natural blockers of the metastatic process, metastasis suppressors, and identify the mechanisms by which they regulate metastasis. Raf kinase inhibitory protein (RKIP), a protein that regulates kinase activity, is a suppressor of TNBC metastasis. Although RKIP inhibits the activity of key kinases such as Raf-1, GRK2, NIK/IKK in cultured cells, the kinase targets of RKIP in tumors are not known. To address this question, we used a mass spectrometry approach involving inhibitor-conjugated beads to identify kinases that are down-regulated by RKIP in human TNBC xenograft tumors. Our results identified novel targets of RKIP whose kinase activity is either down-regulated or up-regulated by RKIP. We used bioinformatics analysis to build RKIP-regulated signaling networks based upon RKIP-induced changes in kinase activity and related gene expression. Elucidating RKIP function at a systems level reveals the interplay between key metastatic signaling cascades, particularly in relation to cell migration and invasion, and can potentially identify novel anti-metastatic target combinations in TNBCs.

Citation Format: Ali E. Yesilkanal, Casey Frankenberger, Daniel C. Rabe, Gary L. Johnson, Marsha R. Rosner. Elucidating metastatic signaling networks in TNBC by investigating RKIP-regulated kinome. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr B23.
Abstract B29: Dissecting the effects of Brd4 short isoform on breast cancer progression
Authors: Zhu Xinyi , Hunter Kent W.
Published: May 2019

Breast cancer is one of the most common female malignancies in the United States with metastatic disease being the main contributor to breast cancer morbidity and mortality. Our lab previously identified BRD4, a member of the bromodomain family, as a regulator of breast cancer progression. Brd4 is a chromatin-modifying factor that regulates the expression of a large set of genes. Brd4 is normally expressed as two isoforms. The long isoform (Brd4-LF) contains two bromodomains in the N-terminus, an ET domain, a serine-rich SEED domain, and a proline-rich C-terminus. The short isoform (Brd4-SF) differs from the long isoform by a deletion of the proline-rich C-terminus and a three amino acid difference on the N-terminal domain. Previous studies in our laboratory indicate that these two isoforms have opposing effects on breast cancer progression. Highly metastatic mouse mammary tumor cells overexpressing Brd4-LF suppressed tumor growth and dissemination, whereas overexpression of Brd4-SF increased metastatic outgrowth in lungs. We have shown that these two isoforms are found in different compartments within the cell nucleus, leading to altered interaction with nuclear proteins. Brd4-LF is localized in the nuclear matrix while Brd4-SF is found in the nuclear envelope where it interacted with the LINC complex and known metastasis susceptibility proteins RRP1B and SIPA1. Given that these two isoforms reside in different subnuclear locations, we hypothesize that the isoforms bind to different sets of genes. To test this hypothesis we are utilizing the chromatin-immunoprecipitation sequencing (ChIP-seq) assay to characterize the genetic interactions associated with Brd4-SF. Identification of the Brd4-SF interacting genes will provide us with mechanistic insight into the underlying causes of the different metastatic outcomes promoted by the two isoforms, highlighting potential targets for future targeted therapies.

Citation Format: Xinyi Zhu, Kent W. Hunter. Dissecting the effects of Brd4 short isoform on breast cancer progression. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Metastasis; 2015 Nov 30-Dec 3; Austin, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(7 Suppl):Abstract nr B29.