bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3733: Adenine nucleotide translocase2 mediates 18F-FDG uptake in dedifferentiated thyroid cancer
Authors: Lee Chul-Hee , Youn Hyewon , Chung Seock-Jin , Kim Ha , Park Cho Rong , Kim Mi Jeong , Park Young Joo , Cho Sun Wook , Kang Keon Wook , Chung June-Key 
Published: July 2017

Abstract
Objectives: 18F-fluorodeoxyglucose (FDG), an analogue of glucose, provides valuable functional information based on the increased glucose uptake and glycolysis in malignant tumor cells. Although both glucose transporter-1 (GLUT-1) and hexokinase2 (HK2) activity have been considered to associate with FDG uptake, the molecular mechanisms that determine FDG uptake are still largely unknown. Adenine nucleotide translocase 2 (ANT2) in the mitochondria inner membrane was reported to relate with tumor malignancy. We investigated the correlation between FDG uptake and Glut-1, HK2, and ANT2 expression in thyroid cancers throughout various spectrums of differentiation status.
Methods: N-thy-ori (normal human thyroid cells), WRO (follicular cancer), BHP10-3 and TPC-1 (papillary cancer), and FRO (anaplastic cancer) were used for this research. GLUT-1, HK2, and ANT2 expressions were measured by western blot. ANT2 siRNAs and pcDNA3.1-ANT2 vectors were used to modify ANT2 expression. FDG uptakes were measured in thyroid cells and human embryonic kidney cells (293FT) with HK2 or ANT2 transfection. For patient tissue analysis, 95 thyroid tissue-array cores were evaluated, and which are classified 36 as normal, 44 as poorly differentiated (PD), and 15 as anaplastic thyroid cancer (ATC). ANT2 expression was measured by immunostaining, scored from 1 to 5.
Results: FDG uptake in thyroid cancer cells was increased in anaplastic and poorly differentiated cells (P&lt;0.001). GLUT-1 expression was higher in both TPC-1 and FRO than other cells. Whereas, HK2 was expressed only in cancer cells. ANT2 was expressed only in FRO cells and the highest FDG accumulation was also observed in FRO. ANT2 siRNA showed decreased FDG uptake (0.55-fold) and ANT2 overexpression increased FDG uptake (1.7-fold). In 293FT cells, HK2 and ANT2 transfection increased FDG uptake (P&lt;0.01). PD tissues (mean = 41.7, SD = 19.7) and ATC (mean = 48.0, SD = 25.6) tissues from patients showed higher ANT2 expression than normal (P&lt;0.05).
Conclusion: We showed that ANT2 was expressed only in anaplastic thyroid cancer cells, and this was related to FDG uptake. Higher level of ANT2 expression was observed in dedifferentiated cancer, and this indicates that ANT2 can be used as a marker of malignancy in thyroid cancer.
Citation Format: Chul-Hee Lee, Hyewon Youn, Seock-Jin Chung, Ha Kim, Cho Rong Park, Mi Jeong Kim, Young Joo Park, Sun Wook Cho, Keon Wook Kang, June-Key Chung. Adenine nucleotide translocase2 mediates 18F-FDG uptake in dedifferentiated thyroid 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 3733. doi:10.1158/1538-7445.AM2017-3733
Abstract 3747: msa2 biotin-binding CAR combined with biotinylated tumor-specific antibodies or DNA aptamers for universal cancer immunotherapy
Authors: Lohmueller Jason J. , Ham James D. , Finn Olivera J. 
Published: July 2017

Abstract
Chimeric antigen receptor T cells (CAR-Ts) are highly promising as cancer therapeutics, but the creation of CARs for new antigens remains a laborious process. As an alternative strategy researchers are designing universal “tag-CARs” that instead of directly recognizing tumor antigens, recognize tags on tumor-specific antibodies bound to tumor cells, thereby activating T cell effector functions. Here we report the creation of a novel tag-CAR that uses the high affinity monomeric streptavidin 2 (msa2) biotin binding domain to target cancer cells labeled with biotinylated tumor-specific antibodies or DNA aptamers. We constructed lentiviral vectors encoding msa2 CARs with three different signaling architectures - CD28-zeta, CD28-OX40-zeta, and 4-1BB-zeta. After confirming that each msa2 CAR was efficiently expressed on the surface of Jurkat and primary human T cells, we found that plate-immobilized biotin was capable of specifically inducing T cell activation markers and cytokine production. We then stained a variety of cancer cell lines with various tumor-specific biotinylated antibodies or DNA aptamers targeting tumor antigens such as MUC1, CD19, CD20, and EGFR and found that the stained cells specifically activated the msa2-CAR cells and led to cytokine production. We are currently investigating the ability of msa2-CAR cells to lyse antibody and aptamer-bound cancer cells and will be assessing their ability to shrink tumors in a human tumor xenograft mouse model.
Citation Format: Jason J. Lohmueller, James D. Ham, Olivera J. Finn. msa2 biotin-binding CAR combined with biotinylated tumor-specific antibodies or DNA aptamers for universal cancer 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 3747. doi:10.1158/1538-7445.AM2017-3747
Abstract 3751: Allogeneic EGFRvIII chimeric antigen receptor T cells for treatment of glioblastoma
Authors: Wong Oi Kwan , Dusseaux Mathilde , Ma Jing-Tyan , Au Melinda , Leduc Sophie , Chou Joyce , Yu Jessica M. , Bateman Marjorie , Pertel Thomas , Lindquist Kevin C. , Smith Julianne , Sasu Barbra , Liu Shu-Hui 
Published: July 2017

Abstract
Glioblastoma multiforme (GBM) is a highly aggressive form of brain tumors with a 5-year survival rate of less than 10%. Standard-of-care combining radiation therapy with temozolomide only yields a median survival of 14.6 months and more effective therapeutic options to extend patient lives are urgently needed. EGFR variant III (EGFRvIII) is a tumor specific mutant of EGFR found in 25-30% of GBM but not expressed in healthy tissues. EGFRvIII is formed by an in-frame deletion of exons 2-7 of the wild-type EGFR which leads to removal of 267 amino acids in the extracellular domain of the receptor. The truncated receptor loses its ability to bind ligands but acquires constitutive kinase activity. The lack of normal tissue expression makes EGFRvIII an ideal target for developing CAR T therapy. Most CAR T therapies in clinical development use the patients’ own T cells for CAR T manufacturing. Our approach is to develop an “off-the-shelf” CAR-T treatment for GBM using healthy donor T cells with the TCR (T-cell receptor) disrupted to prevent graft-versus-host disease. This allogeneic approach could circumvent challenges faced by some patients due to limited availability and quality of their own T cells and rapid progress of their diseases, and has the potential to reduce the high cost associated with autologous CART therapy. Using phage panning and hybridoma approaches, we generated a series of humanized and fully human EGFRvIII antibodies with a wide range of affinities. Recombinant EGFRvIII protein binding by Biacore and FACS assays using EGFRvIII and EGFR wild-type expressing cells demonstrated that these antibodies are highly specific for EGFRvIII. The antibodies were converted to ScFvs and cloned into a CAR vector. Subsequently, EGFRvIII CARs were transduced into primary human T cells for functional studies. We developed a series of assays to evaluate CAR expression, degranulation activity and target dependent cytotoxicity. Our goal is to select CAR candidate that is safe, persistent and has potent target dependent cytotoxicity.
Citation Format: Oi Kwan Wong, Mathilde Dusseaux, Jing-Tyan Ma, Melinda Au, Sophie Leduc, Joyce Chou, Jessica M. Yu, Marjorie Bateman, Thomas Pertel, Kevin C. Lindquist, Julianne Smith, Barbra Sasu, Shu-Hui Liu. Allogeneic EGFRvIII chimeric antigen receptor T cells for treatment of glioblastoma [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 3751. doi:10.1158/1538-7445.AM2017-3751
Abstract 3757: Ewing Sarcoma regression by Allo-MHC/Chm1 specific T cells without GVHD
Authors: Thiel Uwe , Schober Sebastian , Einspieler ingo , Kirschner Andreas , Schirmer David , Grünewald Thomas , Lüttichau Irene Teichert-von , Richter Guenther , Sorensen Poul , Burdach Stefan 
Published: July 2017

Abstract
Background: Chondromodulin-I (CHM1) sustains malignancy in Ewing sarcoma (ES). Refractory ES carries a dismal prognosis and patients with bone marrow (BM) metastases do not survive irrespective of therapy. We assessed HLA-A*02:01/ CHM1 specific allorestricted T cell receptor (TCR) wildtype and transgenic cytotoxic (CD8+) T cells against ES.
Patients and Methods: Three refractory HLA-A2+ ES patients were treated with HLA-A*02:01/peptide specific allorepertoire-derived (i.e. allorestricted) CD8+ T cells. Patient #1 received up to 4.8x105/kg body weight HLA-A*02:01- allorestricted donor derived wild type CD8+ T cells. Patient #2 received up to 8.2x106/kg HLA-A*02:01- donor derived and patient #3 up to 6x106/kg autologous allorestricted TCR transgenic CD8+ T cells. All patients were treated with the same TCR complementary determining region 3 allorecognition sequence for CHM1319.
Results: HLA-A*02:01/CHM1319 specific allorestricted CD8+ T cells showed specific in vitro lysis of all patient-derived ES cell lines. Therapy was well tolerated and did not cause graft versus host disease (GVHD). Patients #1 and #3 showed slow progression, whereas patient #2, while having BM involvement, showed partial metastatic regression associated with T cell homing to involved lesions. CHM1319 TCR transgenic T cells could be tracked in his BM for weeks.
Conclusions: CHM1319-TCR transgenic T cells home to affected BM and may cause partial disease regression. HLA-A*02:01/antigen specific allorestricted T cells proliferate in vivo without causing GVHD.
Citation Format: Uwe Thiel, Sebastian Schober, ingo Einspieler, Andreas Kirschner, David Schirmer, Thomas Grünewald, Irene Teichert-von Lüttichau, Guenther Richter, Poul Sorensen, Stefan Burdach. Ewing Sarcoma regression by Allo-MHC/Chm1 specific T cells without GVHD [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 3757. doi:10.1158/1538-7445.AM2017-3757
Abstract 3769: Stem cell-engineered invariant natural killer T cells for cancer therapy
Authors: Yang Lili , Smith Drake , Liu Siyuan , Ji Sunjong , Liu Bo 
Published: July 2017

Abstract
Invariant natural killer T (iNKT) cells comprise a small population of αβ T lymphocytes. They bridge the innate and adaptive immune systems and mediate strong and rapid responses to many diseases, including cancer, infections, allergies and autoimmunity. However, the study of iNKT cell biology and the therapeutic applications of these cells are greatly limited by their small numbers in vivo (~0.01-1% in mouse and human blood). Here, we report a new method to generate large numbers of iNKT cells in mice through T cell receptor (TCR) gene engineering of hematopoietic stem cells (HSCs). We showed that iNKT TCR-engineered HSCs could generate a clonal population of iNKT cells. These HSC-engineered iNKT cells displayed the typical iNKT cell phenotype and functionality. They followed a two-stage developmental path, first in thymus and then in the periphery, resembling that of endogenous iNKT cells. When tested in a mouse melanoma lung metastasis model, the HSC-engineered iNKT cells effectively protected mice from tumor metastasis. This method provides a powerful and high-throughput tool to investigate the in vivo development and functionality of clonal iNKT cells in mice. More importantly, this method takes advantage of the self-renewal and longevity of HSCs to generate a long-term supply of engineered iNKT cells, thus opening up a new avenue for iNKT cell-based immunotherapy.
Citation Format: Lili Yang, Drake Smith, Siyuan Liu, Sunjong Ji, Bo Liu. Stem cell-engineered invariant natural killer T cells for cancer therapy [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 3769. doi:10.1158/1538-7445.AM2017-3769
Abstract 3770: CAR-T cell harboring a camelid single domain antibody as a targeting agent to kill tumors expressing VEGFR2
Authors: Chao Heman , Tian Baomin , Uger Marni , Wong Wah 
Published: July 2017

Abstract
Modulation of the immune system is showing tremendous promise in the treatment of malignancies. In addition to checkpoint inhibitors that re-activate T cells present in the tumor microenvironment, exogenously transduced chimeric antigen receptor (CAR) T cells are providing excellent responses in clinical trials for the treatment of leukemias. In this study, we describe CAR-T cells that target VEGFR2-expressing tumors.
Angiogenesis is the process of new blood vessel formation and is essential for a tumor to grow beyond a certain size. Tumors secrete the pro-angiogenic factor vascular endothelial growth factor (VEGF), which acts upon local endothelial cells by binding to vascular endothelial growth factor receptors (VEGFR). As VEGFR2 is also expressed by a variety of tumors, we investigated the utility of anti-VEGFR2 CAR-T cells as a method to treat VEGFR2-expressing tumors.
Camelid antibodies are small (14 kD) single chain antibodies. To generate a camelid antibody targeting the extracellular domain of VEGFR2, a llama was immunized with recombinant VEGFR2/Fc. A phage display library was generated and screened to identify an antibody with high binding affinity to VEGFR2. The selected antibody was expressed in the E. coli. BL21 (DE3) pT7 system. The purified antibody was characterized by SEC, LC-MS peptide mapping and ELISA.
CAR-T cells were engineered to express the camelid anti-VEGFR2 antibody in combination with the CD28 and 4-1BB costimulatory molecules and the CD3 zeta chain. Tumor cells were screened for expression of VEGFR2, and the HL-60 acute promyelocytic leukemia, ZR-75-30 breast ductal carcinoma and NCI-H23 non-small cell lung adenocarcinoma were identified. Co-incubation of anti-VEGFR2 CAR-T cells with the VEGFR-2-expressing cell lines resulted in dose-dependent target cell toxicity as measured by LDH release. In addition, T cell activity was confirmed, as high levels of IL-2 and IFN-γ were detected in the cell culture media. These results suggest that anti-VEGFR2 CAR-T may be useful in directly targeting VEGFR2-expressing tumors.
We previously showed the utility of camelid antibodies in CAR-T constructs as anti-CEACAM6 CAR-T cells show both in vitro and in vitro efficacy against the pancreatic tumor Bx-PC3. The use of the a camelid V21 antibody to target VEGFR2-expressing tumors provides hope that camelid single domain antibodies can be developed for CAR-T therapies.
Citation Format: Heman Chao, Baomin Tian, Marni Uger, Wah Wong. CAR-T cell harboring a camelid single domain antibody as a targeting agent to kill tumors expressing VEGFR2 [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 3770. doi:10.1158/1538-7445.AM2017-3770
Abstract 3771: Concurrent GVAX and anti-OX40 therapy in murine intracranial glioma model
Authors: Jahan Nusrat , Talat Hammad , Curry William T. 
Published: July 2017

Abstract
Introduction: Malignant glioma is the most aggressive brain tumor with a median survival time of less than 1.5 years post diagnosis despite standard treatments. However, recent advancements in immunotherapy hold significant promise. Irradiated whole tumor cell vaccination stimulates antitumor immunity and prolongs survival in mice with intracranial glioma tumors. We have shown that vaccination and immune checkpoint blockade is synergistic and these efforts are underway in the clinic. In contrast to immune checkpoint blockade, agonist stimulation of OX40, a member of the TNFR/TNF superfamily that is expressed on activated CD4 and CD8 T lymphocytes increases T cell proliferation, clonal memory differentiation, and cytokine release. We hypothesized that subcutaneous vaccination with irradiated GM-CSF expressing glioma cells (GVAX) would, therefore, synergize with systemic delivery of stimulating anti-OX40 antibody and provide effective immunity against intracranial gliomas.
Methods: Gl261 cells were implanted in the right frontal lobes of syngeneic c57/bl6 mice using a stereotactic implantation technique. Mice received concurrent subcutaneous injection of irradiated GM-CSF expressing glioma cells and intraperitoneal injection of OX40 antibody, either monotherapy, or control treatment on days 3, 6 and 9 following tumor implantation.
Results: Combination immunotherapy with irradiated GM-CSF expressing glioma cells and intraperitoneal delivery of agonist monoclonal antibody against OX40 was more effective than either therapy alone and prolonged survival in mice with established intracranial GL261 tumors. Elispot analyses of peripheral lymphocytes revealed that combination immunotherapy reverses the Th2 cytokine skew that is induced by the presence of intracranial glioma. OX40 signaling alone and in combination with vaccination drove high levels of interferon gamma expression by splenocytes harvested from glioma-bearing animals, while IL-10 expression was little changed. In the tumor microenvironment, vaccination improves the CD8+ /FoxP3+ T lymphocyte ratio, while OX40 ligation overcomes tumor-driven lymphocyte exhaustion, demonstrated by reduced expression of PD1 and LAG3, measured by flow cytometery 2 weeks after initiation of treatment.
Conclusions: Combination treatment with GVAX and OX-40 agonist antibody increased survival in mice bearing intracranial gliomas and are mechanistically complementary.
Citation Format: Nusrat Jahan, Hammad Talat, William T. Curry. Concurrent GVAX and anti-OX40 therapy in murine intracranial glioma model [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 3771. doi:10.1158/1538-7445.AM2017-3771
Abstract 3773: Tapping CD4 T cells for cancer immunotherapy
Authors: Wälchli Sébastien , Myhre Marit R. , Mensali Nadia , Fåne Anne , Lislerud Kari , Kvalheim Gunnar , Gaudernack Gustav , Inderberg Else M. 
Published: July 2017

Abstract
T-cell based immunotherapy represents an attractive strategy for the treatment of cancer. Whereas cellular anti-tumor immune responses have typically been attributed to CD8 T cells, CD4 T cells play a critical role in tumor elimination and the priming and maintenance of CD8 T-cell responses. Recent findings have highlighted new opportunities for CD4 T cells in cancer immunotherapy. We have isolated CD4+ T cells reactive against tumor antigens from patients who experienced clinical benefit from treatment with cancer vaccines targeting antigens such as hTERT, survivin and frequent neoantigens such as frameshift mutated TGFβRII. Strong T-cell responses against the vaccine or unrelated cancer antigens suggesting epitope spreading correlated with enhanced survival and tumor regression in late stage cancer patients. These HLA class II restricted T-cell clones recognised target cells loaded with long peptides or protein and for some CD4+ T cell clones we could also show direct tumor recognition. TCRs were expressed in expanded third-party T cells by mRNA electroporation or retroviral transduction and tested for functionality. Both CD8+ and CD4+ T cells expressing the TCRs produced TNF-α, IFN-γ and redirected T cells had the capacity to kill following co-incubation with their targets. Selecting highly functional CD4+ T-cell clones reactive against tumor-associated or -specific antigens from patients with clinical responses after treatment with immunotherapy is a successful method for identifying highly functional HLA class II restricted TCRs for adoptive immunotherapy. These HLA class II-restricted TCRs may be of therapeutic value both in haematopoietic malignancies and in melanoma where tumor cells frequently express HLA class II. Furthermore, combining HLA class I- and class II-restricted TCRs for T-cell redirection may provide a more potent therapeutic effect in adoptive T cell therapy.
Citation Format: Sébastien Wälchli, Marit R. Myhre, Nadia Mensali, Anne Fåne, Kari Lislerud, Gunnar Kvalheim, Gustav Gaudernack, Else M. Inderberg. Tapping CD4 T cells for cancer 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 3773. doi:10.1158/1538-7445.AM2017-3773
Abstract 3782: Genetic analysis using a novel high-purity enrichment system for circulating tumor cells independent of epithelial cell antigen
Authors: Takagi Hidenori , Kozuka Masahiro , Ito Hiroshi , Kim Soo Hyeon , Hirai Mitsuharu , Fujii Teruo 
Published: July 2017

Abstract
Background
Genetic analysis of circulating tumor cells (CTCs) is useful as a liquid biopsy. However, there are 3 challenging issues in processing of CTC samples for clinical use of the analysis: [1] numerous residual blood cells in processed samples, [2] loss of CTCs that do not express epithelial markers, and [3] very laborious process. Here, we developed a novel system capable of overcoming all of these problems.
Methods
Our CTC analysis system is composed of 3 steps: [1] filtering of whole blood followed by immunostaining and magnetic labeling of cells trapped on the filter, [2] depletion of white blood cells (WBCs) in the cells recovered from the filter by magnetic separation, and [3] trapping the resultant cells at an observation chamber in a microfluidic enrichment device using dielectrophoresis, followed by recovering them as a concentrated sample after fluorescence microscope observation. SNV and CNV analysis was conducted using the collected concentrated sample, and the system performance was substantiated. Genetic mutations of the cells in the collected sample were detected by the Quenching Probe method. On the other hand for CNV analysis, cells in the collected concentrated sample were immobilized on slide glass and analysis of gene amplification conducted by FISH.
Results
With SNV analysis, our system successfully detected EGFR, KRAS or PIK3CA mutations of cancer cell lines spiked in 8 mL of whole blood (Table). The detection sensitivity of our method was 1 cell/mL, and both the cancer cells and their genetic mutations were detected within 9 hours of starting the processing of whole blood. Additionally, with CNV analysis, HER2 gene amplification was confirmed with cell lines spiked in 8 mL of blood. This detection result was obtained within 48 hours.
Conclusion
Our system would be useful for the analysis of gene mutations in a wide range of CTC types independent of certain epithelial antigens, such as CK and EpCAM, and can be used for various genetic analyses.
Table Results of spiking test for SNV analysisSpiked cell linesExpected count[cells/8ml]No. of detected spiked cells [cells]No. of detected remained WBCs [cells]Detected genotypesNCI-H197512979EGFR L858RNCI-H16508225EGFR exon19 deletionSW6208648KRAS codon12, 13SNU-1124154KRAS codon12, 13MCF-777341PIK3CA G1633A
Citation Format: Hidenori Takagi, Masahiro Kozuka, Hiroshi Ito, Soo Hyeon Kim, Mitsuharu Hirai, Teruo Fujii. Genetic analysis using a novel high-purity enrichment system for circulating tumor cells independent of epithelial cell antigen [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 3782. doi:10.1158/1538-7445.AM2017-3782