bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3153: Critical roles of chemo-resistant effector and regulatory T cells in cancer immunotherapy during hemostatic proliferation
Authors: Sato Ko , Watanabe Satoshi , Saida Yu , Tanaka Tomohiro , Baba Junko , Ohtsubo Aya , Shoji Satoshi , Ishikawa Daisuke , Kondo Rie , Okajima Masaaki , Miura Satoru , Tanaka Junta , Kagamu Hiroshi , Yoshizawa Hirohisa , Narita Ichiei 
Published: August 2015

Abstract
Antitumor immunity has been well established to be augmented by cytotoxic lymphodepletion therapies. Adoptively transferred naïve and effector T cells proliferate extensively and show enhanced antitumor effects during homeostatic proliferation when they were adoptively transferred into tumor-bearing hosts that were lymphodepleted with cytotoxic agents or by whole body irradiation. Although the impact of lymphodepletion on transferred donor T cells has been well evaluated, its influence on recipient T cells is largely unknown. The current study demonstrates that both regulatory T cells (Tregs) and effector CD8+ T cells from lymphopenic recipients play critical roles in the development of antitumor immunity after lymphodepletion. Cyclophosphamide (CPA) treatment depleted lymphocytes more efficiently than other cytotoxic agents, such as fludarabine, cisplatin, etoposide, paclitaxel or gemcitabine; however, the percentage of CD4+CD25+Foxp3+ Tregs was significantly increased in CPA-treated lymphopenic mice. Depletion of these chemo-resistant Tregs following CPA treatment and transfer of naïve CD4+ T cells augmented the antitumor immunity and significantly suppressed tumor progression. Further analyses revealed that recipient CD8+ T cells were responsible for this augmentation. Using Rag2−/− mice or depletion of recipient CD8+ T cells after CPA treatment abrogated the augmentation of antitumor effects in CPA-treated reconstituted mice. The transfer of donor CD4+ T cells enhanced the proliferation of CD8+ T cells and the priming of tumor-specific CD8+ T cells originating from the lymphopenic recipients. These results highlight the importance of the recipient cells surviving cytotoxic regimens in cancer immunotherapies.
Citation Format: Ko Sato, Satoshi Watanabe, Yu Saida, Tomohiro Tanaka, Junko Baba, Aya Ohtsubo, Satoshi Shoji, Daisuke Ishikawa, Rie Kondo, Masaaki Okajima, Satoru Miura, Junta Tanaka, Hiroshi Kagamu, Hirohisa Yoshizawa, Ichiei Narita. Critical roles of chemo-resistant effector and regulatory T cells in cancer immunotherapy during hemostatic proliferation. [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 3153. doi:10.1158/1538-7445.AM2015-3153
Abstract 3154: Identification of CD4+ T cell epitopes specific for the breast cancer associated antigen NY-BR-1
Authors: Gardyan Adriane , Osen Wolfram , Agawal Maria , Zörnig Inka , Ruggiero Eliana , Schmidt Manfred , Schneeweiss Andreas , Jäger Dirk , Eichmüller Stefan B. 
Published: August 2015

Abstract
Questions: Whether CD4+ T cell epitopes specific for the breast cancer associated antigen NY-BR-1 can be identified using HLA-transgenic (HLAtg) mice.
Introduction: Adoptive transfer of TCR-transduced autologous tumor antigen-specific T cells provides an innovative strategy for cancer immunotherapy. The differentiation antigen NY-BR-1 is over-expressed in approx. 60% of all invasive mammary carcinomas, thus representing a potential target for cancer immunotherapy. We thus screened NY-BR-1 for the presence of CD4+ T cell epitopes.
Methods: Splenocytes from DNA immunized HLA-DRB1*0301 and HLA-DRB1*0401 tg mice were screened ex vivo by IFNγ ELISpot assay against a NY-BR-1-derived peptide library. In silico predicted candidate epitopes present among recognized library peptides were used to establish CD4+ T cell lines as a read tool to prove HLA-DR-restriction and natural processing of these epitopes by human cells. NY-BR-1-specific CD4+ T cell reactivity among PBMC of breast cancer patients was tested after long term in vitro restimulation with synthetic 15mers by intracellular cytokine staining.
Results: Three HLA-DRB1*0301-restricted and four HLA-DRB1*0401-restricted NY-BR-1-specific CD4+ T cell epitopes were identified in HLAtg mice. HLA-DR-restriction was confirmed upon specific recognition of peptide-loaded T2/DR3 and T2/DR4 target cells by epitope-specific CD4+ T cell lines established from HLAtg mice. These T cell lines specifically recognized human dendritic cells loaded with cell lysates from Ad5-NY-BR-1 infected melanoma cells, showing natural processing of the epitopes. Moreover, upon deep sequencing of TCRs from these murine CD4+ T cell lines two NY-BR-1-specific TCRs were identified which could be considered for the generation of autologous TCR-transduced T cells lines. Finally, CD4+ T cells reactive against the NY-BR-1 specific epitopes were also detected among PBMCs of HLA-DR matched breast cancer patients.
Conclusion: NY-BR-1-specific, HLA-DRB1*0301 and HLA-DRB1*0401 restricted CD4+ T cell epitopes could be identified using HLAtg mice and CD4+ T cells reactive against these epitopes are present among PBMCs of breast cancer patients. Finally, murine HLA-restricted CD4+ T cell lines might serve as source of NY-BR-1-specific TCRs for adoptive immunotherapy approaches.
Citation Format: Adriane Gardyan, Wolfram Osen, Maria Agawal, Inka Zörnig, Eliana Ruggiero, Manfred Schmidt, Andreas Schneeweiss, Dirk Jäger, Stefan B. Eichmüller. Identification of CD4+ T cell epitopes specific for the breast cancer associated antigen NY-BR-1. [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 3154. doi:10.1158/1538-7445.AM2015-3154
Abstract 3156: Construction and validation of an activating and inhibitory chimeric antigen receptor (CAR) system
Authors: Chicaybam Leonardo , Bonamino Martin H. 
Published: August 2015

Abstract
Introduction: Chimeric antigen receptors (CARs) consists a promising type of adoptive immunotherapy. CARs consist of an antigen recognition unit (scFv), a transmembrane (TM) region and an intracellular activation domain. However, on target/off tumor responses due to target recognition in normal cells limits the widespread application of this therapy. In B-cell precursor acute lymphoblastic leukemia, lymphocytes expressing anti-CD19 CARs (19BBz) are being used in the clinical setting. However, the expression of CD19 as a pan-B marker can lead to undesired side effects such as depletion of mature B cells. As mature B cells express both CD19 and CD20 antigens, we propose the creation of an inhibitory anti-CD20 CAR, that when used in combination with 19BBz would be able to discriminate between leukemic blasts and normal B cells. Methods and results: Three inhibitory CARs were constructed containing signaling domains of CTLA-4, PD-1 or BTLA. Jurkat T cells expressing the plasmid pGL4.30 - expressing luciferase controlled by a NFAT responsive promoter - were generated. K562 cell line was used as target cells and modified to express CD19 (K5-19), CD20 (K5-20) or CD19 and CD20 (K5-19/20). In coculture experiments, Jurkat cells expressing 19BBz showed induction of luciferase activity when cultured with K5-19 or K5-19/20. However, Jurkat expressing both activation and inhibitory CAR showed inhibition of luciferase activity when incubated with K5-19/20 while maintained high activity when cultured with K5-19 cells. Furthermore, all three inhibitory CARs were able to inhibit the expression of the activation marker CD69 induced by 19BBz. In primary human T lymphocytes, the 20PD1 CAR did not inhibit proliferation or cytotoxicity induced by 19BBz. Surprisingly, T cells expressing 19BBz and 20PD1 showed preferential lysis of NALM6 CD19+ CD20+ cells, suggesting that the addition of PD1 domain may increase T-cell function in specific contexts. This effect was partially reversed by using a mutant PD1 domain where the tyrosines were replaced by phenylalanine and was completely reversed by using an anti-CD20 CAR truncated at the TM region, suggesting that this phenomenon is dependent of signaling. Conclusions: Additional experiments are needed to evaluate the activity of 20PD1 in primary human T cells. Experiments with inhibitory CARs 20LAG3 and 20PD1LAG3 are ongoing and can serve as an alternative to CAR 20PD1 in the construction of a conditional response system.
Citation Format: Leonardo Chicaybam, Martin H. Bonamino. Construction and validation of an activating and inhibitory chimeric antigen receptor (CAR) system. [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 3156. doi:10.1158/1538-7445.AM2015-3156
Abstract 3168: A role for tumor-derived CCL2 in directing leukocyte infiltration and stromal heterogeneity in pancreatic ductal adenocarcinoma
Authors: Tooker Graham M. , Gladney Whitney L. , Beatty Gregory 
Published: August 2015

Abstract
Tumor heterogeneity within solid tumors is a recognized barrier to the efficacy of standard therapies. In pancreatic ductal adenocarcinoma (PDAC) the microenvironment is characterized by varying degrees of cytokine/chemokine production, stromal extracellular matrix, and immune cell infiltrates (Hezel et al. 2006). To investigate stromal phenotypical differences in PDAC, we have used the KrasG12D-; Trp53R172H; Pdx-1 Cre (KPC) genetically engineered mouse model which recapitulates tumor heterogeneity seen in human PDAC (Hingorani et al. 2005). Gross histological analysis of KPC tumors allowed us to stratify tumors into 2 broad groups: stromal and non-stromal. We found that stromal tumors were comprised of a significant macrophage infiltrate, whereas non-stromal tumors were generally void of macrophages. We modeled this stromal heterogeneity using tumor cell lines derived from KPC tumors. We found that, when implanted under the skin of wild-type mice, some cell lines produced stromal tumors rich in macrophages, while others formed non-stromal tumors with few macrophages within the microenvironment. Supernatant cultured from these KPC tumor cell lines in vitro revealed elevated CCL2 production, specifically in cell lines from stromal tumors. As CCL2 is a known chemo-attractant for monocytes, we hypothesized that tumor-derived CCL2 contributes to monocyte recruitment and stromal heterogeneity within the tumor microenvironment. To test this hypothesis, we have used an in vitro trans-well migration assay. With this assay, we found that more bone marrow derived cells migrate toward supernatant from stromal tumors relative to non-stromal tumors. This finding was lost in the presence of a CCL2 inhibitor. Ongoing studies are examining a role for tumor-derived CCL2 in regulating tumor-associated stroma in PDAC. Our findings suggest the importance of tumor-immune cell interactions in defining stromal heterogeneity and may offer insight into mechanisms that regulate the stromal compartment in human PDAC.
Citation Format: Graham M. Tooker, Whitney L. Gladney, Gregory Beatty. A role for tumor-derived CCL2 in directing leukocyte infiltration and stromal heterogeneity in pancreatic ductal adenocarcinoma. [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 3168. doi:10.1158/1538-7445.AM2015-3168
Abstract 3186: Role of Interleukin 1 signaling in tumor elicited inflammation and colon cancer
Authors: Dmitrieva Oxana , Possoco David , Francescone Ralph A. , Hou Vivianty , Vendramini-Costa Debora B. , Grivennikov Sergey 
Published: August 2015

Abstract
Colorectal cancer (CRC) is the 2nd most common cause of cancer deaths in the United States and other developed countries, despite important advances in detection, surgery and chemotherapy. It becomes increasingly clear that not only cell autonomous events in cancer cells are required for tumor growth and progression. Growing CRC tumors possess the ability to recruit inflammatory cells and upregulate inflammatory cytokines - a phenomenon recently introduced as “Tumor elicited inflammation”(TEI). The mechanisms of TEI induction and tumor promotion are not yet well defined. IL23/IL17 pathway is one of the established regulators of TEI. While in mice the ablation of IL-23 signaling results in a uniform loss of IL-17 response and in a decrease in tumor growth; in humans IL23/IL-23R gene expression signature is not always consistent with elevated IL-17 signature, implicating that there should be other regulators of IL-17 and TEI induction in tumors. IL-1 pathway is important for the induction of T helper IL-17 producing cells (Th17) and for the production of IL-17 by innate lymphoid cells (ILC), both in mice and in humans. IL-1 pathway is also implicated in host defense, immunity and tumorigenesis, with some of its functional characteristics similar to IL-23 pathway. In addition, IL-1, much alike IL-23, can be also induced by various microbial derived and host derived ‘danger signals’ readily available within the tumor microenvironment. Our own data suggests that components of IL-1 pathway are upregulated in CRC tumors. Therefore, we hypothesized, that IL-1 pathway might be an important regulator of TEI and CRC tumorigenesis. Using genetic tools and animal models, we examined if IL-1 has the potential role in different stage of CRC and whether it serves as an important regulator of TEI and CRC tumor cells proliferation and survival.
Citation Format: Oxana Dmitrieva, David Possoco, Ralph A. Francescone, Vivianty Hou, Debora B. Vendramini-Costa, Sergey Grivennikov. Role of Interleukin 1 signaling in tumor elicited inflammation and colon cancer. [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 3186. doi:10.1158/1538-7445.AM2015-3186
Abstract 3202: Single cell analysis of ascites macrophages in ovarian cancer
Authors: He Huanhuan Mahsa , Yu Feiqiao Brian , Quake Stephen R. , Dorigo Oliver 
Published: August 2015

Abstract
Macrophages compose a major part of the tumor microenvironment (TME) in several types of cancer, including ovarian cancer [1]. Studies have shown that a high percentage of tumor associated macrophages (TAMs) correlates with a poor prognosis in ovarian cancer [2], due to the crucial role of TAMs in promoting tumor growth and angiogenesis [3,4]. Yet not much investigation has been executed to reveal the function of macrophages in ovarian cancer ascites, a hallmark of ovarian cancer.
Previous gene expression analysis in TAMs often used bulk samples [5,6]. However, due to the complexity of the peritoneal environment, this approach had the effect of “dampening” rare and potentially informative signals from a heterogeneous population such as ascites macrophages. Therefore, we resorted to single-cell expression analysis to dissect the heterogeneity of ascites macrophages.
Recent breakthroughs in single cell RNA-seq techniques have made it more cost effective to process a large number of samples at single cell resolutions. Here we used Fluidigm's Single-Cell Auto Prep System, a state-of-art technology for automate single cell cDNA synthesis [7]. We sequenced single ascites macrophages from ID8 murine ovarian cancer model and single alternatively activated bone marrow derived macrophages (BMDM-M2). When comparing the single cell expression profiles of ascites macrophages to BMDM using principle component analysis (PCA), we observed that the ascites macrophages scattered broadly and could be further divided into two groups, whereas BMDM were tightly clustered into one group. Furthermore, gene oncology (GO) analysis of highly expressed genes in each group revealed that ascites macrophages had a distinct gene expression profile from BMDM, highlighting the metabolic pathways, especially catabolic process. We are currently working on validating these findings in human patients with ovarian carcinoma. In summary, this work sheds light on the understanding of the role of ascites macrophages in ovarian cancer.
1. Qian BZ, Pollard JW. Cell. 2010;141:39-51.
2. He YF, Zhang MY, Wu X, et al. PLoS One. 2013;8:e79769.
3. Mantovani A, Germano G, Marchesi F, et al. Eur J Immunol 2011;41:2522-25.
4. Murray PJ, Wynn TA. Nat Rev Immunol, 2011;11:723-37.
5. The Cancer Genome Atlas Research Network. Nature. 2014;511:543-50.
6. The Cancer Genome Atlas Research Network. Nature. 2014;513:202-9.
7. Wu AR, Neff NF, Kalisky T, et al. Nat Methods. 2014;11:41-6.
Citation Format: Huanhuan Mahsa He, Feiqiao Brian Yu, Stephen R. Quake, Oliver Dorigo. Single cell analysis of ascites macrophages in ovarian cancer. [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 3202. doi:10.1158/1538-7445.AM2015-3202
Abstract 3253: Metastasis suppression by non-KP54 kisspeptins
Authors: Hampton Kelsey R. , Pounds Keke M. , Tremblath Andrew P. , Welch Danny R. 
Published: August 2015

Abstract
The KISS1 metastasis suppressor blocks metastasis by inhibiting colonization at secondary sites. KISS1 activity requires secretion and extracellular processing by furin into kisspeptins. Kisspeptin-54 (KP54; G68 to F121) binds a G-protein coupled receptor, KISS1R, and the mechanism of metastasis suppression has been assumed to be through the receptor. However, KISS1 is cleaved into kisspeptins outside of the cell, and KISS1R expression is not required for metastasis suppression. Therefore, an alternate hypothesis is that a non-KISS1R binding kisspeptin(s) is responsible for metastasis suppression. To test this hypothesis, we engineered all ten of the possible kisspeptins predicted from known dibasic cleavage sites (M1-Q145; M1-R56; M1-R67; M1-R124; R56-R67; G68-F121; E55-F121; E55-Q145; G68-Q145; E125-Q145) downstream of the KISS1 secretion signal peptide (designated KMP1 to KMP10, respectively) and expressed each individually in B16-F10 melanoma cells. Expression was confirmed by immunoblot and/or RT-QPCR analysis. Following intravenous injection, experimental lung metastases were quantified. KP54 expression suppressed metastasis. Some KMP lacking the KISS1R ligand suppressed metastasis as well as KP54 and full-length KISS1 (p&lt;0.05). Specifically, the N-terminal kisspeptin (M1 - R56) dramatically suppressed metastasis. This suggests that the mechanism of action for metastasis suppression by KISS1 does not necessarily require activation of KISS1R by the peptide ligand but also leaves the possibility for additional receptors. Alternatively, KISS1/KMP interactions with other intracellular proteins may be required for metastasis suppressor activity. Support: CA134981, Natl Fndn Cancer Res, Komen SAC11037, KS Bioscience Auth, P30-CA168524, Steiner Family Fund.
Citation Format: Kelsey R. Hampton, Keke M. Pounds, Andrew P. Tremblath, Danny R. Welch. Metastasis suppression by non-KP54 kisspeptins. [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 3253. doi:10.1158/1538-7445.AM2015-3253
Abstract 3255: The role of Hdj2 protein in metastatic progression of experimental intracranial glioma
Authors: Meshalkina Darya A. , Shevtsov Maxim A. , Margulis Boris A. , Guzhova Irina V. 
Published: August 2015

Abstract
Glioblastoma is one of the most malignant cancer types with median survival of 15 months regardless of the application of combined radio- and chemotherapy. Expression of Hdj2 negatively correlates with the increase of tumor aggressiveness. For assessment of Hdj2 role in tumor progression, invasiveness and metastasis formation in gliomas we developed three C6-based cell lines with Hdj2 and related proteins knock-down: C6 shHsp70, C6 shHdj1 and C6 shHdj2 itself. Following intracranial injection of the modified tumor cells the animals’ survival was estimated. As compared to control C6 (25.4±3.9 days) and C6 shHdj1 (25.5±3.8) we observed a nearly 1.5-fold decrease in survival in C6 shHdj2 (16.8±3.5 days) (P&lt;0.05). On the contrary, in C6 shHsp70 group the survival increased up to 42.5±12.0 days (the increase is completely explainable by the slower growth rate of the culture). Subsequent MR imaging and histological analysis of the tumors demonstrated elevated invasiveness and metastatic activity in C6 shHdj2 group in comparison to C6 shHsp70, C6 shHdj1 and control C6. High migration activity and the ability of detached C6 shHdj2 cells to settle on the substrate was proved in wound-healing assay, colony forming assay and calcium-dependent cell aggregation assay. Immunofluorescence analyses showed loss of membrane-expressed N-cadherin and loss of intercellular contacts mediated by N-cadherin in C6 shHdj2 cells in comparison to other considered cell lines. Actin staining with rhodamine-falloidin revealed highly abundant leading edges in C6 shHdj2 culture.
Decrease of Hdj2 expression in gliomas has proved to be of high importance in tumor progression and formation of metastasis.
Citation Format: Darya A. Meshalkina, Maxim A. Shevtsov, Boris A. Margulis, Irina V. Guzhova. The role of Hdj2 protein in metastatic progression of experimental intracranial glioma. [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 3255. doi:10.1158/1538-7445.AM2015-3255
Abstract 3257: NME1 mediates a switch in expression of beta integrin subunits that correlates with prolonged patient survival
Authors: Leonard M. Kathryn , Novak Marian , Yang Xiuwei , Belkin Alexey , Kaetzel David 
Published: August 2015

Abstract
Expression of the metastasis suppressor NME1 in melanoma is associated with reduced cellular motility and invasion in vitro and metastasis in vivo, but the molecular mechanisms underlying this activity are not completely understood. Herein we report a novel mechanism through which NME1 modulates focal adhesion dynamics via regulation of integrin β1. Stable expression of NME1 significantly altered focal adhesion turnover at the cell periphery. Interestingly, over-expression of NME1 resulted in a switch in from predominantly fast recycling α4β1 integrins to slower recycling αvβ3 integrins. Contrary to its regulation of other cell surface receptors, the inhibition of integrin β1 by NME1 was found to occur at the transcriptional level rather than through dynamin-mediated endocytosis. This effect of NME1 required both its 3-5′ exonuclease and nucleoside diphosphate kinase (NDPK) activities, which are also required for its metastasis suppressor activities in vivo. Furthermore, an inverse correlation was observed between NME1 and integrin β1 mRNA in a large cohort of primary melanoma biopsies. The inverse correlation of NME1 and integrin β1 RNA was also a strong predictor of prolonged distant disease-free and overall survival in patients with the basal-like subtype of breast carcinoma. Together, these data strongly suggest NME1 suppresses metastasis of human melanoma and some types of breast cancers by inhibiting integrin β1 expression, which reduces recycling of focal adhesions and suppresses cell motility.
Citation Format: M. Kathryn Leonard, Marian Novak, Xiuwei Yang, Alexey Belkin, David Kaetzel. NME1 mediates a switch in expression of beta integrin subunits that correlates with prolonged patient survival. [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 3257. doi:10.1158/1538-7445.AM2015-3257