bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3937: Diversity index as a novel predictor of tumor progression in breast cancer
Authors: Chung Yul Ri , Kim Hyun Jeong , Kim Young A , Chang Mee Soo , Hwang Ki-Tae , Park So Yeon 
Published: July 2017

Abstract
Intratumoral genetic heterogeneity leads to tumor progression and therapeutic resistance. However, due to the difficulty associated with its assessment, application of intratumoral genetic heterogeneity as a prognostic or predictive marker is limited. We investigated the significance of Shannon diversity index as a tool for measuring genetic heterogeneity by performing fluorescence in situ hybridization of c-Myc gene in invasive breast cancers, and correlated the Shannon diversity index for c-Myc copy number variation with clinicopathologic features of tumor including patient survival. Shannon index for c-Myc copy number variation strongly correlated with average c-Myc copy number and was significantly higher in tumors with c-Myc genetic or regional heterogeneity than in those without c-Myc amplification. High Shannon index was associated with high histologic grade, lymphovascular invasion, p53 overexpression, high Ki-67 proliferation index, hormone receptor negativity, and HER2 amplification. In survival analyses, c-Myc amplification or c-Myc copy number gain was not associated with patient survival. However, a high level of Shannon index was associated with poor disease-free survival. In subgroup analyses, it was found to be an adverse prognostic factor in hormone receptor-positive group, but not in hormone receptor-negative group. In a validation set, high Shannon diversity index for c-Myc copy number variation was also found to be associated with poor survival of the patients. We further investigated the correlation with clinicopathologic features and predictive power of Shannon index using another gene, fibroblast growth factor receptor 1 (FGFR1) and observed that high Shannon index for FGFR1 gene copy number variation was an independent prognostic factor for poor clinical outcome in a whole group and in hormone receptor-positive subgroup. In conclusion, this study demonstrated that higher diversity index was associated with adverse pathologic parameters of breast cancer and poor clinical outcome suggesting that Shannon diversity index, which represents intratumoral genetic heterogeneity, can be used as a potential biomarker for tumor progression and prognostication in patients with breast cancer.
Citation Format: Yul Ri Chung, Hyun Jeong Kim, Young A Kim, Mee Soo Chang, Ki-Tae Hwang, So Yeon Park. Diversity index as a novel predictor of tumor progression in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3937. doi:10.1158/1538-7445.AM2017-3937
Abstract 3954: Single-cell misregulation of NRF2 in basal-like breast cancer
Authors: Pereira Elizabeth , Janes Kevin 
Published: July 2017

Abstract
Cancer is driven by a complex and heterogeneous combination of genes and pathways, many of which are not explicitly oncogenic on their own. These so-called non-oncogenes are required for maintenance of the tumorigenic state, but they are not mutated themselves. One way to identify “non-oncogene” pathways is to evaluate how proliferating cells adjust to variable and evolving environments. By examining heterogeneous transcriptional regulatory states of single cells in a 3D culture model of breast epithelial organization, we uncovered NRF2 as a candidate transcription factor upstream of an important regulatory state for morphogenesis. NRF2 is a known oncogene in lung cancer, but mutations in the transcription factor are uncommon in breast cancer. To examine the functional impact of NRF2 signaling, we engineered several triple-negative breast cancer cell lines with inducible short hairpin RNA to knock down NRF2. In claudin-low MDA-MB-231 cells, knockdown of NRF2 caused dramatic reduction in stellate invasion compared with shGFP control. In basal-like HCC-1806 cells, knockdown of NRF2 caused a reduction in “ruptured” tumorspheres, in which cells disseminate from their originating site. In a non-transformed breast epithelial cell line, overexpression of a non-degradable form of NRF2 caused greater variation in the size of acini, while knockdown of NRF2 narrowed the size distribution of acini. Therefore, knockdown of NRF2 substantially normalizes the size and shape of both non-transformed and cancerous 3D structures. We also analyzed expression levels of the genes in the provisional NRF2-regulated cluster after NRF2 knockdown or overexpression. Upon NRF2 knockdown, the p53-regulated transcripts CDKN1A (p21) and RPS27L were strongly induced, suggesting that p53 may be activated after NRF2 loss. Our results suggest a functional importance for NRF2 in multiple breast-epithelial and TNBC contexts, and raise the possibility of interplay between NRF2 and p53. A detailed understanding of NRF2 regulation and dynamics could enable new strategies to disrupt this non-oncogene addiction and re-sensitize cancer cells to their environment and to chemotherapy.
Citation Format: Elizabeth Pereira, Kevin Janes. Single-cell misregulation of NRF2 in basal-like breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3954. doi:10.1158/1538-7445.AM2017-3954
Abstract 3960: Tumor cell drug resistance induced by the programmed death ligand 1 (PD-L1) immune checkpoint is associated with autophagy
Authors: Minassian Lori M. , MacDonald-Goodfellow Shannyn K. , Truesdell Peter , Sanwalka Daniel , Craig Andrew W. , Koti Madhuri , Siemens D Robert , Graham Charles H. 
Published: July 2017

Abstract
The interaction between the Programmed Death Ligand 1 (PD-L1) immune checkpoint on the tumor cell surface with the Programmed Death-1 (PD-1) receptor on cytotoxic T lymphocytes (CTLs) leads to CTL inactivation, thereby promoting tumor cell escape from adaptive immunity. We previously demonstrated that signaling by PD-L1/PD-1 is bidirectional and leads to activation of oncogenic pathways as well as drug resistance in tumor cells. We also have preliminary evidence that Immunity Related GTPase M, an important mediator of autophagy, is up-regulated by PD-1/PD-L1 reverse signaling. Autophagy is a well-established mechanism of drug resistance in cancer cells. This led us to hypothesize that PD-1/PD-L1 signaling induces drug resistance in tumor cells by up-regulating autophagy. The MEK/ERK and the PI3K/Akt signaling pathways are known to increase and decrease autophagy, respectively. Breast cancer cells exposed to rPD-1 showed a time dependent increase in extracellular signal–regulated kinase (ERK) activation and a decrease in protein kinase B (Akt) activation. Conversion of microtubule-associated protein light chain 3 (LC3)-I to LC3-II is a requirement for autophagosome formation and is a robust marker of autophagy. Exposure of human breast cancer cells to recombinant PD-1 (rPD-1) showed a time-dependent increase in LC-3 II. We are currently conducting additional studies to confirm that the activation of PD-L1 signaling in tumor cells up-regulates autophagy. These results provide evidence that PD-1/PD-L1 reverse signaling activates autophagy as a potential mechanism of cancer cell chemoresistance. (Supported by a grant from the Canadian Institutes of Health Research.)
Citation Format: Lori M. Minassian, Shannyn K. MacDonald-Goodfellow, Peter Truesdell, Daniel Sanwalka, Andrew W. Craig, Madhuri Koti, D Robert Siemens, Charles H. Graham. Tumor cell drug resistance induced by the programmed death ligand 1 (PD-L1) immune checkpoint is associated with autophagy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3960. doi:10.1158/1538-7445.AM2017-3960
Abstract 3972: Ex vivo profiling of PD-1 blockade using organotypic tumor spheroids
Authors: Barbie David A. , Jenkins Russell , Wong Kwok-Kin 
Published: July 2017

Abstract
Despite the impressive activity of PD-1/PD-L1 immune checkpoint blockade in melanoma, lung, and other cancers, the majority of patients are intrinsically resistant to this therapy. Furthermore, despite years of effort few robust biomarkers of response exist, and tumor PD-L1 expression and mutational load are only partially predictive of activity. Nearly all attempts at biomarker development have relied on fixed tumor tissue or studies in plasma. Meanwhile, emerging functional approaches to study patient derived samples, such as organoid development and circulating tumor cell culture, lack features of the native tumor immune microenvironment and thus are limited in the ability to assess ex vivo anti-PD-1 response. We have developed a novel approach to study short-term ex vivo response to immune checkpoint blockade based upon short term culture of murine- and patient-derived organotypic tumor spheroids (MDOTS/PDOTS) in a 3-dimensional microfluidic system. Spheroids derived from fresh mouse and human tumor samples through partial collagenase digestion and filtration retain autologous tumor infiltrating immune cells, including PD-1 positive CD8 T lymphocytes. Using murine syngeneic mouse cancer models with known responsiveness or resistance to PD-1 blockade, we demonstrate the ability to recapitulate tumor killing ex vivo or lack thereof. Multiplexed cytokine profiling of conditioned media in this system further enables characterization of the cytokine and chemokine response to PD-1 and/or CTLA-4 blockade. We thus analyzed the secreted cytokine/chemokine response to PD-1 inhibition across a large panel of PDOTS, and observed pronounced induction of CCL19/CXCL13, which associated with % PD-1 positive CD8 T cells. This strategy further uncovered production of a class of myeloid attracting chemokines and cytokines in major proportion of samples, which clustered with the presence of plasmacytoid dendritic cells and myeloid derived suppressor cells. Importantly, the presence of this maladaptive cytokine/chemokine signature correlated with lack of response in the subset of patients that had been treated with PD-1 blockade. Together, these data demonstrate the ability to profile the functional response to tumor PD-1 blockade ex vivo and unveil a novel strategy to advance precision immuno-oncology.
Citation Format: David A. Barbie, Russell Jenkins, Kwok-Kin Wong. Ex vivo profiling of PD-1 blockade using organotypic tumor spheroids [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3972. doi:10.1158/1538-7445.AM2017-3972
Abstract 3977: Neutrophils fertilize the pre-metastatic omental niche in ovarian cancer
Authors: LEE WonJae , Naora Honami 
Published: July 2017

Abstract
More than 60% of ovarian cancer patients present with peritoneal carcinomatosis that almost always involves the omentum, a fat pad that suspends from the stomach. However, the mechanisms that promote the tropism of ovarian cancer cells for the omentum are poorly understood. Because the omentum is the major peritoneal conduit and reservoir for immune cells, we hypothesized that immune cells control the propensity of ovarian cancer cells to implant on the omentum. In this study, we evaluated changes in immune cell populations in the omentum and in other visceral fat pads in the peritoneal cavity in mouse xenograft models of ovarian cancer. Intriguingly, the proportion of neutrophils was observed to dramatically increase in the omentum prior to tumor cell implantation at this site. On the other hand, the proportion of neutrophils did not substantially increase in other visceral fat pads. Furthermore, the proportion of macrophages did not significantly increase in any of the fat tissues prior to tumor cell implantation. Collectively, these findings raise the possibility that early recruitment of neutrophils to the pre-metastatic omental niche promotes a microenvironment that is conducive to ovarian cancer cell implantation at this site.
Citation Format: WonJae LEE, Honami Naora. Neutrophils fertilize the pre-metastatic omental niche in ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3977. doi:10.1158/1538-7445.AM2017-3977
Abstract 3981: The Shc1 scaffold protein simultaneously balances Stat1 and Stat3 activity in breast cancer to promote immune suppression and resistance to immunotherapy
Authors: Ahn Ryuhjin , Jacquet Kévin , Fabian Marc , Huang Sidong , Bisson Nicolas , Ursini-Siegel Josie 
Published: July 2017

Abstract
Receptor and cytoplasmic tyrosine kinases are key signal integrators in poor outcome breast cancers that are central to the establishment of an immunosuppressive microenvironment. Immunotherapies represent an emerging approach within the armament of anti-cancer agents. Although the efficacy of tyrosine kinase inhibitors relies, in part, on their ability to augment adaptive immunity, the increased heterogeneity and functional redundancy of the tyrosine kinome in poor outcome breast cancers represents a significant hurdle to achieving durable responses to immunotherapies. We previously identified the Shc1 (ShcA) scaffold protein, a central regulator of tyrosine kinase signaling, as essential for promoting immune suppression. We recently showed that the ShcA pathway simultaneously activates STAT3 immunosuppressive signals and impairs STAT1-driven immune surveillance in breast cancer cells. Impaired phosphorylation of select tyrosine residues on ShcA potently and selectively reduces STAT3 activation in breast tumors, profoundly sensitizing them to immune checkpoint inhibitors and tumor vaccines. Meanwhile, impaired phosphorylation of other select tyrosine residues on ShcA potently increased antigen presentation and sensitivity to tumor vaccines in preclinical mouse models. Based on these results, we have set out to elucidate protein interactors dependent on distinct ShcA phospho-tyrosines to regulate STAT1 and STAT3 signaling axis that aid tumor driven immune suppression. We combined affinity-purification mass spectrometry (AP/MS) and proximity-dependent biotin identification (BioID) followed by mass spectrometry (BioID/MS). Known interactors of ShcA as well novel interactors have been identified. Promising candidates that could be critical in immune suppression downstream of ShcA have been validated to be true interactors by co-IP and BioID assays. They have been screened for immune regulation in vitro for further modulation in vivo. Currently, inhibitors of phospho-tyrosine motifs of ShcA do not exists. Therefore, the development of pharmacological inhibitors to prevent phospho-tyrosine ShcA dependent STAT3 signaling or relieve suppression of STAT1 signaling may be an attractive method to strategically sensitize breast tumors to multiple immunotherapies.
Citation Format: Ryuhjin Ahn, Kévin Jacquet, Marc Fabian, Sidong Huang, Nicolas Bisson, Josie Ursini-Siegel. The Shc1 scaffold protein simultaneously balances Stat1 and Stat3 activity in breast cancer to promote immune suppression and resistance to immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3981. doi:10.1158/1538-7445.AM2017-3981
Abstract 3995: Identification of receptor tyrosine kinases (RTKs) expression in tumor-infiltrating lymphocytes (TILs) and peripheral blood mononuclear cells (PBMC)
Authors: Khochenkov Dmitry , Volkova Maria , Tsimafeyeu Ilya 
Published: July 2017

Abstract
Aim of this study is to discover expression levels of RTKs (VEGFR1, VEGFR2, PDGFRa, PDGFRb) in CD45+ population of PMBC and TILs isolated from patients with clear cell renal cell cancer (RCC). Here we present preliminary data.
Species of tumors and blood were prospective collected from 9 patients with RCC after radical nephrectomy or tumor resection from July to October 2016. TILs isolation was performed under standard protocol with slight modification. Briefly, tumor was shredded with scissors on small pieces of 2-3 mm and twice washed in RPMI 1640 media by centrifugation for 10 min at 1500 rpm. After centrifugation, pellet was diluted RPMI 1640 with enzyme mix (Collagenase and Dispase II) on Petri dish and placed on shaker-thermostat andincubate sample for 12h at 37C with continuous shaking. After incubation, cell suspension was passed thought a cell strainer, mesh size 70 μm, placed on a 50 mL tube and washed twice washed in RPMI 1640 media by centrifugation.
For TILs isolation cell suspension was placed on Ficoll-Paque Plus and isolated by centrifugation for 25 min at 1500 rpm. Cell pellet was resuspended in RPMI 1640 media. PMBC was isolated from whole blood using the Ficoll-Paque method. Isolated TIL and PMBC were prepared for flow cytometry at typical protocol with or without permeabilization.
Cells were stained with anti-CD45 FITC-conjugated antibodies (Novus), primary anti-VEGFR1, anti-VEGFR2, anti-PDGFRa, anti-PDGFRb (Abcam) mouse antibodies and Alexa-568 conjuated secondary anti-mouse antibodies (Invitrogen) and analyzed on BD Canto II flow cytometer.
Expression of RTKs on CD45+ cell population were evaluated. Among PMBC 86±9% of cells was CD45-positive and expressed RTKs (10±1% cells were positive for VEGFR1 and VEGFR2, and 6±2% cells were positive for PDGRFa and PDGFRb). TILs contained 65±18% of CD45-positive cells and did not express RTKs on the cell surface (less than 1% of cells), but intracellular staining showed presence of RTKs: 8±2% of cells were VEGFR1 or VEGFR2-positive, 20±8% of cells were PDGFRa or PDGFRb-positive.
Preliminary data demonstrate extracellular expression of RTKs on PBMC and intracellular expression of RTKs in TILs. The study will be continued.
Citation Format: Dmitry Khochenkov, Maria Volkova, Ilya Tsimafeyeu. Identification of receptor tyrosine kinases (RTKs) expression in tumor-infiltrating lymphocytes (TILs) and peripheral blood mononuclear cells (PBMC) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3995. doi:10.1158/1538-7445.AM2017-3995
Abstract 4003: Acral lentiginous melanoma and infiltrating lymphocytes in a Latin American population
Authors: Castaneda Carlos , Torres-Cabala Carlos , Casavilca Sandro , Castillo Miluska , Villegas Valeria , Sanchez Joselyn , Flores Claudio , Gomez Henry , Vidaurre Tatiana 
Published: July 2017

Abstract
Background: Acral lentiginous melanoma (ALM) is unusual in Caucasian but is the most common subtype in Hispanic populations, and has been associated to poor prognosis. Herein we compare clinicopathological features associated to ALM in a Latin-American population and prospectively evaluate its association with tumor infiltrating lymphocytes (TIL).
Patients and Methods: All patients with melanoma presenting from 2005 to 2012 at Instituto Nacional de Enfermedades Neoplasicas in Lima, Peru, were retrospectively identified. Analysis of association between clinicopathological features and ALM was performed. Survival analysis compared the outcome of ALM to whole group and extremity NALM. We performed prospective evaluation of TIL following Azimi et al recently presented classification.
Results: A total of 537 ALM and 287 NALM cases were identified. Older age (p= 0.02), thicker Breslow (p= 0.008), positive lymph node (p= 0.051) and ulceration (p&lt;0.001) were found to be more frequent in ALM. Acral had worse OS compared with both whole group (p= 0.04) and extremity NALM (p &lt;0.001). Stage I-II patients had a median OS of 5.3 (95%CI: 4.3-6.2) for ALM and 9.2 (95% CI: 5.0 - 7.0) for extremity NALM (p=0.016). TIL was prospectively evaluated in 490 cases, and Grade 0-1, II and III of TIL represented 44.7%, 33.6% and 27.1%, respectively. Higher TIL grade was associated to less Breslow thickness (p= 0.021), less mitosis index and NAM. Higher TIL grade was also associated to better OS (p&lt; 0.001)
Conclusion: ALM is highly prevalent in Latin-America and carry poor outcome. Lower TIL levels was associated to poor outcome and ALM.
Citation Format: Carlos Castaneda, Carlos Torres-Cabala, Sandro Casavilca, Miluska Castillo, Valeria Villegas, Joselyn Sanchez, Claudio Flores, Henry Gomez, Tatiana Vidaurre. Acral lentiginous melanoma and infiltrating lymphocytes in a Latin American population [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4003. doi:10.1158/1538-7445.AM2017-4003
Abstract 4014: Blood-brain-barrier spheroids: Next-generation screening platform for brain-penetrating agents
Authors: Cho Choi-Fong 
Published: July 2017

Abstract
The inability of most systemically delivered therapeutics to cross the blood-brain-barrier (BBB) is considered a major barrier to effective brain cancer treatment. In malignant glioma the tumor neovasculature is known to be leaky, however invasive cells remain protected behind an intact BBB, and continue to thrive within the brain, ultimately leading to tumor regrowth and patient death. This leaves us with an urgent unmet need for the development of next-generation therapeutics with improved brain delivery. For the first time, we describe here the utility of 3D multicellular BBB spheroids made of human brain endothelial cells (ECs), pericytes and astrocytes as a screening tool for brain-penetrating agents. We show that the outer surface of the spheroids, composed primarily of ECs and pericytes, form a tight barrier which is permeabilized in the presence of VEGF. The barrier is characterized by the presence of intact tight junctions and efflux-pump activity (i.e., P-glycoprotein). Furthermore, we have used this model to successfully demonstrate the transport of angiopep-2 (a well-known brain delivery vector) and its conjugates (containing cargoes of various sizes such as peptide, protein and affibody), thereby displaying the versatility of this model to screen and study a wide range of therapeutic agents. We demonstrate that this model is superior to the conventional transwell model in maintaining essential BBB characteristics (i.e., tight/adherens junctions and P-glycoprotein expression) and as a drug-screening tool. We have utilized the spheroid model to screen a panel of cell-penetrating peptides (CPPs) to identify several candidates with high brain-penetration potential. We then verified the ability of the top 4 candidate CPPs to cross the BBB in mice. This high-throughput model can lead to better design and analysis of first-in-class glioma therapeutics, and improve prediction of drug penetration in a living model, paving the way for breakthrough discoveries in brain cancer.
Citation Format: Choi-Fong Cho. Blood-brain-barrier spheroids: Next-generation screening platform for brain-penetrating agents [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4014. doi:10.1158/1538-7445.AM2017-4014