bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4024: Understanding the role of mitochondria in the progression from normal fibroblasts to DCIS associated fibroblasts
Authors: Ko Ying-Hui , Lin Zhao , Martinez Outschoorn Ubaldo E. 
Published: August 2015

Abstract
Background and Purpose: Ductal carcinoma in situ (DCIS) is a non-invasive breast cancer which may progress to invasive breast cancer. Many studies have focused on the epithelial cell mechanisms of progression from normal breast tissue to DCIS. However, in this study we want to study mechanisms of progression in cancer associated fibroblasts from normal breast tissue to DCIS.
Loss of caveolin 1 (CAV1) in fibroblasts is a marker of reduced mitochondrial membrane potential and is a prognostic biomarker in ductal carcinoma in situ. High mitochondrial membrane potential in epithelial cells is a crucial determinant of carcinogenesis.
Methods: In the study we use MCF10DCIS as a model for DCIS. MCF10DCIS.com (MCF10DCIS) is a clonal breast cancer cell line. The MCF10A, non-tumorigenic epithelial cell line, and human immortalized but non-transformed dermal fibroblasts (BJ1) were used study the difference between non-tumorigenic epithelial cells and DCIS and interactions between these two epithelial cells and fibroblasts. In order to study the interaction between epithelial cells and fibroblasts, we set up a coculture model system comprised both epithelial cells and fibroblasts. Immunofluorescence staining and confocal microscopy were used here to visualize the specific protein expression in different cell types.
Results: (1) MCF10DCIS cells in coculture with fibroblasts have increased mitotracker red staining while as fibroblasts have decreased mitotracker staining compared to homotypic culture. However, coculture of MCF10A cells with fibroblasts did not change the mitotracker staining in either MCF10A or fibroblasts. (2) Loss of stromal CAV1 protein expression is observed in fibroblasts when fibroblasts are cocultured with MCF10DCIS cells compared to homotypic culture. CAV1 protein expression is unchanged in coculture with MCF10A cells.
Conclusion: Previous studies have demonstrated that loss of stromal CAV1 and changes in mitochondrial membrane potential are factors involved in the transition from DCIS to invasive breast cancer. In this study we show that loss of stromal CAV1 and increases in epithelial mitochondrial membrane potential may play a role in the progression to DCIS.
Citation Format: Ying-Hui Ko, Zhao Lin, Ubaldo E. Martinez Outschoorn. Understanding the role of mitochondria in the progression from normal fibroblasts to DCIS associated fibroblasts. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4024. doi:10.1158/1538-7445.AM2015-4024
Abstract 4036: Palmitoylated Ras-driven MAPK signaling, transformation and tumorigenesis, but not tumor progression, are spatially regulated by plasma membrane microdomains
Authors: Michael James , Wurtzel Jeremy G.T. , Goldfinger Lawrence E. 
Published: August 2015

Abstract
In this study, we assessed the contributions of plasma membrane (PM) microdomain targeting to the functions of H-Ras and paralogue R-Ras. Activated H-Ras is a mitogen which can drive tumorigenesis through Raf activation, while R-Ras is weakly mitogenic and does not induce the MAPK pathway in cells, despite binding Raf in vitro. These paralogues have identical effector-binding regions, but variant C-terminal targeting domains (tDs). Ras tDs are responsible for lateral distribution within PM microdomains as a result of distinct lipid modifications: activated H-Ras targets to lipid ordered/disordered (Lo/Ld) domain borders, whereas active R-Ras is sequestered in Lo domains (rafts). We hypothesize that PM segregation regulates effector interactions and downstream signaling. We used tD swap mutants, and assessed effects on MAPK pathway activation, cell proliferation, transformation, and tumorigenesis. Activated R-Ras harboring the H-Ras tD (R-Ras-tH), stably expressed in NIH3T3 fibroblasts, interacted with Raf and induced MEK and ERK phosphorylation. Furthermore, R-Ras-tH stimulated cell growth, and supported anchorage-independent growth in soft agar, indicating induction of a transformed phenotype in vitro. Conversely, stable transfectants of H-Ras harboring the R-Ras tD (H-Ras-tR) had reduced MEK and ERK phosphorylation, cell growth, and colony formation in soft agar compared with H-Ras cells. Thus, Ras access to Raf at the PM is a critical step for Ras mitogenic signaling. However, tumor progression driven by these cells in xenografts in mice did not match in vitro phenotypes. R-Ras-expressing cells produced few solid tumors, while R-Ras-tH cells produced relatively larger, but size-limited, avascular tumors with a higher frequency per xenograft. Surprisingly, both H-Ras and H-Ras-tR xenografts produced large and heavily vascularized tumors, implicating an important role of Ras in modulating tumor angiogenesis in this model. Indeed, the conditioned media (secretome) of R-Ras-expressing cells did not support endothelial cell cord formation in growth factor-reduced Matrigel, while the secretome from H-Ras cells robustly induced cord formation; these effects were independent of Ras microdomain localization. Thus, targeting Ras to the Lo/Ld border is sufficient to initiate tumor formation, however, a tD-independent angiogenic effect is required for progression of Ras-induced tumors.
Citation Format: James Michael, Jeremy G.T. Wurtzel, Lawrence E. Goldfinger. Palmitoylated Ras-driven MAPK signaling, transformation and tumorigenesis, but not tumor progression, are spatially regulated by plasma membrane microdomains. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4036. doi:10.1158/1538-7445.AM2015-4036
Abstract 4039: Inhibition of Pak4 blocks growth of breast cancer cells
Authors: Rane Chetan K. , Senapedis William , Baloglu Erkan , Shacham Sharon , Minden Audrey G. 
Published: August 2015

Abstract
The p21-activated kinase family of proteins promotes cell survival and plays an important role in cell proliferation, cell cycle regulation and cell shape determination. There are six mammalian PAK proteins which can be subdivided into two groups by sequence homology and mode of activation- Group A Paks consisting of Pak 1, 2 and 3 and Group B Paks consisting of Pak 4, 5 and 6. There is a growing list of evidence that PAK proteins are overexpressed in many cancer types, including colon, lung, and breast cancer. PAK4 is frequently overproduced in breast cancer, including Her2 positive and triple negative breast cancers, while it is expressed at low levels in normal mammary tissue. Our preliminary evidence showed that untransformed mouse mammary epithelial cells have low levels of PAK4 that become elevated when these cells are transformed by Her2 overexpression. Furthermore, PAK4 overexpressed in mammary epithelial cells leads to oncogenic transformation and tumorigenesis in mice. We hypothesize that PAK4 plays an important role in promoting mammary tumorigenesis, and that it can serve as an effective drug target for breast cancer treatment. To test this hypothesis, we have analyzed two orally available PAK4 Allosteric Modulators (PAM; KPT-8752 and KPT-9274), which reduce the steady state PAK4 protein level in cancer cells. We observed that when we treat breast cancer cells with KPT-8752, we severely block the cell growth. KPT-8752, is effective both in Her2 positive and triple negative breast cancer cells. Our results indicate that Pak4 is an important regulator of breast cancer cell growth. Experiments studying the effects of PAMs (KPT8752 and KPT9274) on tumorigenesis in animal models of breast cancer are ongoing.
Citation Format: Chetan K. Rane, William Senapedis, Erkan Baloglu, Sharon Shacham, Audrey G. Minden. Inhibition of Pak4 blocks growth of breast cancer cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4039. doi:10.1158/1538-7445.AM2015-4039
Abstract 4042: Kruppel-like factor 5 promotes breast cancer proliferation, migration and invasion partially through upregulating the transcription of TNFα-induced protein 2
Authors: Jia Lin , Liu Rong , Chen Ceshi 
Published: August 2015

Abstract
The Krupple-like factor 5 (KLF5) transcription factor is highly expressed in high-grade and basal-like TNBC breast cancers. Our previous studies reported that KLF5 promotes breast cancer cell proliferation and survival through target genes, such as FGF-BP and mPGES1. However, the mechanism by which KLF5 promotes cell migration and invasion is still not completely understood. In this study, we demonstrated that TNFAIP2, a TNFα-induced gene, is a KLF5 direct downstream target gene. The expression of the TNFAIP2 is highly correlated with the expression of the KLF5 protein in breast epithelial cell lines at both mRNA and protein levels. Manipulation of KLF5 expression positively alters TNFAIP2 mRNA and protein expression levels in multiple breast cell lines. KLF5 binds to TNFAIP2 gene promoter and activates its transcription. Functionally, TNFAIP2 promotes cell proliferation and xenograft growth in NOD-SCID mice. Importantly, knockdown of either KLF5 or TNFAIP2 significantly reduces cancer cell migration and invasion. Overexpression of TNFAIP2 rescues KLF5 knockdown-induced migration suppression in HCC1937 cells. TNFAIP2 interacts with two small GTPase Cdc42 and Rac1, increases their activities, and changes actin skeleton and cell morphyology. Finally, KLF5 and TNFAIP2 promote cell migration and invasion through activating Cdc42 and Rac1. These findings collectively suggest that TNFAIP2 is a KLF5 direct target gene and both KLF5 and TNFAIP2 promotes TNBC cell migration and invasion through Cdc42 and Rac1.
Citation Format: Lin Jia, Rong Liu, Ceshi Chen. Kruppel-like factor 5 promotes breast cancer proliferation, migration and invasion partially through upregulating the transcription of TNFα-induced protein 2. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4042. doi:10.1158/1538-7445.AM2015-4042
Abstract 4056: Recognition of tumor cells by Dectin-1 orchestrates innate immune cells for anti-tumor responses
Authors: IKUSHIMA Hiroaki , TANIGUCHI Tadatsugu 
Published: August 2015

Abstract
The eradication of cancer cells requires communication between cells which constitute the complex immune system. Natural killer (NK) cells are essential tumor-killing effector cells of the innate arm of the immune system; however, little is known about whether or how other immune cells recognize tumor cells to assist NK cells. In addition, although signal-transducing pattern recognition receptors are known to play important roles in innate immune responses against invading pathogens through recognition of pathogen-associated molecular patterns (PAMPs), it is still enigmatic whether and how these receptors contribute to anti-tumor immune responses. In this study, we show that the innate immune pattern recognition receptor Dectin-1 expressed on dendritic cells (DCs) and macrophages is critical to NK-mediated killing of tumor cells. We also show that tumor cell-mediated Dectin-1 signaling is instigated by the receptor recognition of N-glycan structures on tumor cells, which we term tumor-associated molecular patterns (TAMPs). As such, the TAMP-Dectin-1 signaling causes activation of the Interferon Regulatory Factor 5 (IRF5) transcription factor and subsequent induction of genes required for the full-blown killing activity of NK cells. The importance of these events is underscored by the observation of massive tumor metastasis in mice genetically deficient in either Dectin-1 or IRF5. Thus, these results reveal a hitherto unrecognized facet of pattern recognition receptors in the orchestration of anti-tumor innate immune responses; recognition of TAMPs. This study is the first demonstration that an innate immune pattern recognition receptor potentiates anti-tumor immune responses, offering new insight into anti-tumor activity of the innate immune system with implications for anti-tumor immunotherapy.
Citation Format: Hiroaki IKUSHIMA, Tadatsugu TANIGUCHI. Recognition of tumor cells by Dectin-1 orchestrates innate immune cells for anti-tumor responses. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4056. doi:10.1158/1538-7445.AM2015-4056
Abstract 4058: Hypoxia-induced soluble CD137 in malignant cells blocks CD137L-costimulation as an immune escape mechanism
Authors: Labiano Sara , Palazón Asís , Quetglas José I. , Bolaños Elixabet , Azpilicueta Arantza , Morales-Kastresana Aizea , Rodriguez Alfonso , Rodriguez-Ruiz Maria , Gurpide Alfonso , Aznar M Angela , Jure-Kunkel Maria , Melero Ignacio 
Published: August 2015

Abstract
Hypoxia is a common feature in solid tumors that has been implicated in immune-evasion. In this study, we show that exposure of mouse and human tumor cell lines to hypoxic conditions (1% O2) promotes CD137 transcription. Previous studies from our group have shown that hypoxia up-regulates the co-stimulatory receptor CD137 on activated T lymphocytes and on vascular endothelial cells. However, our results show that the CD137 transcript upregulated under hypoxia in tumor cell lines is predominantly an alternatively spliced form that encodes for a soluble variant, which lacks the transmembrane domain. Accordingly, soluble CD137 (sCD137) is detectable by ELISA in the supernatant of hypoxia-exposed cell lines and in the serum of tumor-bearing mice. sCD137, as secreted by tumor cells, is able to bind to CD137-Ligand (CD137L). Our studies on primed T lymphocytes in co-culture with stable transfectants for CD137L demonstrate that tumor-secreted sCD137 prevents co-stimulation of T lymphocytes. Such an effect results from preventing the interaction of CD137L with the transmembrane forms of CD137 expressed on T lymphocytes undergoing activation. This mechanism is interpreted as a molecular strategy deployed by tumors to repress lymphocyte co-stimulation via CD137/CD137L.
Citation Format: Sara Labiano, Asís Palazón, José I. Quetglas, Elixabet Bolaños, Arantza Azpilicueta, Aizea Morales-Kastresana, Alfonso Rodriguez, Maria Rodriguez-Ruiz, Alfonso Gurpide, M Angela Aznar, Maria Jure-Kunkel, Ignacio Melero. Hypoxia-induced soluble CD137 in malignant cells blocks CD137L-costimulation as an immune escape mechanism. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4058. doi:10.1158/1538-7445.AM2015-4058
Abstract 4059: Crosstalk between CD8+ T and NK cells: fine-tuning of antitumor immune response
Authors: Uzhachenko Roman V. , Goodwin J Shawn , Costa Lino , Terekhov Alexander , Thounaojam Menaka C. , Hofmeister William H. , Shanker Anil 
Published: August 2015

Abstract
Local tumor antigen-specific T cell-NK cell collaboration is indispensable for the elimination of tumor cells, including antigen-deficient tumor escape variants before metastasis. While mechanistic details are available for the innate instruction of the T cell responses, little is known for the adaptive control of NK cell activity. We observed in a mouse model of mastocytoma expressing a self tumor antigen P1A that effector CD8+ T cells provided a necessary “help” to dormant NK cells in eliciting their antitumor effector function. Bioluminescence imaging of mastocytoma tumors following adoptive transfer of P1A-specific T cells in RAG-/- and RAG-/-γc-/- mice showed that NK cell anti-tumor activity requires cytolytic T cells, whereas T cells can function independent of NK cells. In 2D and 3D co-culture systems, we observed that PMA/ionomycin-stimulated CD8+ T cells form multiple contacts with naïve NK lymphocytes. Data show that NK cells interacting with activated CD8+ T cells show an up-regulation of CD25 and CD69 expression mediated by intercellular contacts, and activation of NKG2D receptors and Stat2, Stat6, Jak1, Jak3, Tyk2, and PTEN signaling molecules with a decrease in the phosphorylation of Stat1, PKB/Akt, SAPK/JNK, p38. On the other hand, interacting NK cells down-regulate CD25 molecule expression on CD8+ T cells and promote differentiation of central memory CD44+CD62L+ T cells. CD8+ T cells display an elevation in the phosphorylation of Stat1 and down-regulation of Stat5 with stimulated PKB/Akt, Lck, mTOR, and p42/p44. Moreover, significant changes in the cytosolic and mitochondrial Ca2+, production of mitochondrial ROS, mitochondrial membrane potential, mitochondrial permeability transition pore, and synthesis of nitric oxide and non-protein thiols (mostly, reduced glutathion) were observed in a reciprocal T cell-NK cell interaction. These results highlight the importance of mitochondrial activity in the re-modeling of activation signaling and memory differentiation of interacting CD8 T cells and NK cells. These results will help refine cancer immunotherapeutic strategies.
Citation Format: Roman V. Uzhachenko, J Shawn Goodwin, Lino Costa, Alexander Terekhov, Menaka C. Thounaojam, William H. Hofmeister, Anil Shanker. Crosstalk between CD8+ T and NK cells: fine-tuning of antitumor immune response. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4059. doi:10.1158/1538-7445.AM2015-4059
Abstract 4080: Role of nuclear PKM2 in transcriptional regulation leading to EMT
Authors: Konno Masamitsu , Nishida Naohiro , Kawamoto Koichi , Koseki Jun , Doki Yuichiro , Mori Masaki , Ishii Hideshi 
Published: August 2015

Abstract
Pyruvate kinase M2 (PKM2) maintains aerobic glycolysis in cancer cells and stimulates cell proliferation by controlling Warburg effect(Nature, 2008). In addition, PKM2 translocates to nucleus in response to EGFR signaling and up-regulates Cyclin D1 expression by controlling its transcription (Nature, 2011). PKM2 confers malignant potential by controlling both metabolism and transcriptional regulation. In this study, we investigated the correlation between PKM2 and invasion and metastasis of colorectal cancer in terms of nuclear function, which has yet to be investigated. Tumor invasion is initiated with the change of epithelial-mesenchymal transition (EMT) in cancer cell, and we induced EMT by adding TGF-β1 and EGF to SW 480. After EGF induction, in parallel with E-cadherin down-regulation and Vimentin up-regulation, the expression of PKM2 was increased both in transcriptional and translational level and PKM2 translocated into nucleus. PKM2 knock-down by siRNA suppressed E-cadherin decrease and invasion capacity in invasion assay. We created sh pyruvate kinase SW 480 with mouse PKM1 or PKM2 was overexpressed both of which have DNA sequence resistant to sh pyruvate kinase. PKM2 overexpression stimulated EMT induction by enhancing E-cadherin decrease. TGIF2 was identified which interacted with PKM2 in nucleus in response to EMT induction, which was considered to have a key role in controlling EMT induction. We performed immunohistochemical staining with specific antibody to PKM2 using clinical samples of colorectal cancers. Clinicopathological analysis showed that PKM2 positivity significantly correlated with lymph node metastasis and distant organ metastasis. In conclusion, our data suggested that PKM2 nuclear function had a crucial role in controlling invasion and metastasis.
Citation Format: Masamitsu Konno, Naohiro Nishida, Koichi Kawamoto, Jun Koseki, Yuichiro Doki, Masaki Mori, Hideshi Ishii. Role of nuclear PKM2 in transcriptional regulation leading to EMT. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4080. doi:10.1158/1538-7445.AM2015-4080
Abstract 4082: PDLIM2 : A cytoskeleton to nuclear courier protein for the IGF-1, Wnt and TGF beta signalling pathways in Epithelial to Mesenchymal Transition
Authors: Bustamante Garrido Milan , Cox Orla T. , O'Flanagan Ciara , Buckley Deirdre A. , Kiely Patrick A. , O'Connor Rosemary 
Published: August 2015

Abstract
PDLIM2 is a PDZ-LIM domain protein that regulates the stability and activity of transcription factor families (including NFκB, STATs and beta catenin). PDLIM2 expression is repressed in certain cancers but it is also highly expressed in Triple Negative Basal Breast cancers that are characterized by poor survival. Suppression of PDLIM2 reverses the EMT phenotype, inhibits polarized cell migration, and disrupts formation of polarized epithelial acini in 3D cell cultures. PDLIM2 shuttles from the cytoskeleton to the nucleus, but what mediates this nuclear translocation or activity in transcription factor regulation is unknown.
The aim of this study was to identify the mechanisms governing PDLIM2 subcellular localization and nuclear translocation. We found that IGF-1or TGF-β promotes PDLIM2 accumulation in the nucleus. Similarly, WNT3a stimulation enhances PDLIM2 accumulation in the nucleus while inhibition of WNT activity results in PDLIM2 stabilization in the cytoplasm. Cytoplasmic to nuclear translocation is associated with reduced phosphorylation on several serine residues in PDLIM2. The de-phosphorylation and subsequent nuclear translocation of PDLIM2 can be prevented by inhibiting the protein phosphatase PP1. In contrast, PDLIM2 phosphorylation can be enhanced by activation of protein kinase C, which is dependent on the presence of the focal adhesion scaffolding protein RACK1 in a complex with PDLIM2.
Overall, the data indicate that PDLIM2 cytoplasmic to nuclear translocation in response to IGF-1, WNT or TGF beta signalling is mediated by serine phosphorylation and de-phosphorylation by cytoskeleton-associated kinases and phosphatases. Thus PDLIM2 acts as a “cytoskeleton to nucleus” courier protein for these signalling pathways to promote cancer cell migration and EMT.
Citation Format: Milan Bustamante Garrido, Orla T. Cox, Ciara O'Flanagan, Deirdre A. Buckley, Patrick A. Kiely, Rosemary O'Connor. PDLIM2 : A cytoskeleton to nuclear courier protein for the IGF-1, Wnt and TGF beta signalling pathways in Epithelial to Mesenchymal Transition. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4082. doi:10.1158/1538-7445.AM2015-4082