bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract B021: Single domain antibodies targeting CTLA-4 demonstrate a critical role for Fc in the antitumor response
Authors: Dougan Michael , Ingram Jessica , Blomberg Olga , Rashidian Mohammad , Keliher Edmund , Weissleder Ralph , Almo Steven , Ploegh Hidde 
Published: November 2016

Abstract
Monoclonal antibodies targeting CTLA-4 are now a central part of the standard of care for metastatic melanoma, and have shown efficacy against diverse tumor types. However, the extent to which direct blockade of CTLA-4 contributes to the efficacy of these antibodies remains incompletely understood. Mice lacking FcγRIV fail to respond to αCTLA-4 therapy, and maintain robust tumor infiltrating Treg populations. These findings suggest that anti-CTLA-4 antibody therapy enhances antitumor responses through Treg depletion, though a requirement for FcγR in other aspects of the antitumor response has not been excluded, and the importance of CTLA-4 blockade alone has not be directly assessed. We have developed an alpaca-derived single domain antibody (H11) that binds CTLA-4 with high affinity, occluding the B7 binding motif as determined by x-ray crystallography. We are able to use this single domain antibody in 18F immuno-PET to produce a comprehensive image of CTLA-4 expression in a tumor-bearing live mouse, confirming penetration of H11 into the tumor microenvironment. Therapeutically, we find that H11 potently blocks CTLA-4 co-inhibition in vitro, but fails to elicit protective immunity in vivo. Therapeutic activity is restored when H11 is attached to the Fc region of murine IgG2a, but not when half-life alone is increased through H11 dimerization or conjugation to PEG, confirming a central role for Fc-dependent function in αCTLA-4 responses, as well as the minimal efficacy of CTLA-4 blockade alone. We further find that the efficacy of αCTLA-4 therapy can be enhanced through blockade of CD47, consistent with an important role for Treg depletion by myeloid cells in the activity of this therapy. These findings have implications for the development of combinations immune therapies.
Citation Format: Michael Dougan, Jessica Ingram, Olga Blomberg, Mohammad Rashidian, Edmund Keliher, Ralph Weissleder, Steven Almo, Hidde Ploegh. Single domain antibodies targeting CTLA-4 demonstrate a critical role for Fc in the antitumor response [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr B021.
Abstract B031: FAK/PYK2 inhibition enhances antitumor efficacy of anti-PD-1 and anti-4-1BB antibodies
Authors: Pachter Jonathan A. , Weaver David T. 
Published: November 2016

Abstract
Although durable responses to single agent immune checkpoint inhibitors have been reported, additional approaches are needed to extend this therapeutic benefit to a greater proportion of patients. Combination of immunotherapy agents with tumor microenvironment modulators has the potential to overcome barriers that tumor cells develop to evade the immune system, and provide benefit to a greater proportion of patients. Focal Adhesion Kinase (FAK) and the family member PYK2 are potentially valuable targets due to their roles in regulating key cellular populations in the tumor microenvironment. The FAK/PYK2 dual inhibitors, VS-6063 and VS-4718, have previously been shown to inhibit monocyte-derived macrophages, reduce tumor-associated macrophages in xenograft models, and promote a CD8+ T cell-mediated anti-tumor response in squamous cell carcinoma models.
We now report the combination of VS-4718 with an anti-PD-1 mAb shows substantially improved efficacy over anti-PD-1 mAb alone in MC38 syngeneic tumor-bearing animals. Analysis of MC38 tumors at day 12 of treatment revealed a significant increase in the CD8+ T cell/Treg ratio in tumors in the VS-4718 + anti-PD-1 combination group, providing a mechanistic understanding for the enhanced efficacy of this combination.
To explore additional combination options, we tested the combination of VS-4718 with anti-4-1BB in the MC38 model. Consistent with what was observed with the anti-PD-1 combination, addition of VS-4718 also enhanced the efficacy of an anti-4-1BB mAb inducing full tumor regression.
To further delineate the effect of FAK inhibition, an in vitro T cell proliferation assay was conducted. VS-6063 and VS-4718 dose-dependently stimulated proliferation of CD8+ cytotoxic T cells isolated from healthy donors. This is in distinct contrast to other protein kinase inhibitors, such as the SRC inhibitor dasatinib which potently impaired the proliferation of CD8+ cytotoxic T cells. In addition, both VS-4718 and VS-6063 decreased CD8+ T cell exhaustion markers (LAG3 and PD-1), and increased T cell-mediated tumor cell killing in vitro.
These data provide additional rationale for ongoing clinical trials testing the FAK/PYK2 inhibitor VS-6063 in combination with pembrolizumab (anti-PD-1) plus gemcitabine in pancreatic cancer or in combination with avelumab (anti-PD-L1) in ovarian cancer.
Citation Format: Jonathan A. Pachter, David T. Weaver. FAK/PYK2 inhibition enhances antitumor efficacy of anti-PD-1 and anti-4-1BB antibodies [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr B031.
Abstract B037: Macrophage repolarization therapy in metastatic colorectal cancer: CCR5 inhibition
Authors: Halama Niels , Zoernig Inka , Berthel Anna , Kahlert Christoph , Klupp Fee , Suarez-Carmona Meggy , Krauss Juergen , Brand Karsten , Lasitschka Felix , Lerchl Tina , Luckner-Minden Claudia , Ulrich Alexis , Weitz Juergen , Schneider Martin , Buechler Markus W. , Zitvogel Laurence , Herrmann Thomas , Benner Axel , Kunz Christina , Luecke Stephan , Springfeld Christoph , Falk Christine , Jaeger Dirk 
Published: November 2016

Abstract
The influence of the local immune response on the clinical course of colorectal cancer (CRC) has been analyzed extensively. Analyzing the invasive margin of human CRC liver metastases, we identified a protumoral mechanism of T-cell-derived CCL5 that leads to immune cell exploitation by tumor cells. Two distinct subsets of myeloid cells produce CXCL9/CXCL10, which induce an influx of T cells into the invasive margin. CCL5 is produced by these exhausted T cells and stimulates tumor cell proliferation and invasive behavior via CCR5 on tumor cells and macrophages. CCR5 inhibition in patient-derived functional in vitro organotypic culture models induced macrophage repolarization with anti-tumoral effects. These immunomodulatory and anti-tumoral effects of CCR5 blockade then could be confirmed in a phase I trial with a CCR5 antagonist in advanced refractory CRC patients with liver metastases. Amelioration of tumor-promoting inflammation on the tumor tissue level and objective tumor responses in advanced metastatic CRC patients were observed.
Citation Format: Niels Halama, Inka Zoernig, Anna Berthel, Christoph Kahlert, Fee Klupp, Meggy Suarez-Carmona, Juergen Krauss, Karsten Brand, Felix Lasitschka, Tina Lerchl, Claudia Luckner-Minden, Alexis Ulrich, Juergen Weitz, Martin Schneider, Markus W. Buechler, Laurence Zitvogel, Thomas Herrmann, Axel Benner, Christina Kunz, Stephan Luecke, Christoph Springfeld, Christine Falk, Dirk Jaeger. Macrophage repolarization therapy in metastatic colorectal cancer: CCR5 inhibition [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr B037.
Abstract B038: Class IIa HDAC inhibition promotes an antitumor macrophage phenotype that induces breast tumor regression and inhibits metastasis
Authors: Guerriero Jennifer L. , Sotayo Alaba , Ponichtera Holly , Pourzia Alexandra , Schad Sara , Carrasco Ruben , Lazo Suzan , Bronson Roderick , Letai Anthony 
Published: November 2016

Abstract
While tumor-associated macrophages (TAMs) often have net pro-tumor effects, their embedded location and their untapped potential provide impetus to the discovery of strategies to turn them against tumors. We recently reported that a first in class selective class IIa HDAC inhibitor (TMP195) influenced human monocyte responses to colony stimulating factors CSF-1 and CSF-2 in vitro. Here, we utilize a macrophage-dependent autochthonous mouse model of breast cancer to demonstrate that in vivo TMP195 treatment alters the tumor microenvironment and reduces tumor burden and pulmonary metastases through macrophage modulation. TMP195 induces recruitment and differentiation of highly phagocytic and stimulatory macrophages within tumors. Furthermore, combining TMP195 with chemotherapy regimens or T-cell checkpoint blockade in this model significantly enhances the durability of tumor reduction. These data introduce class IIa HDAC inhibition as a novel means to harness the antitumor potential of macrophages to enhance cancer therapy.
Citation Format: Jennifer L. Guerriero, Alaba Sotayo, Holly Ponichtera, Alexandra Pourzia, Sara Schad, Ruben Carrasco, Suzan Lazo, Roderick Bronson, Anthony Letai. Class IIa HDAC inhibition promotes an antitumor macrophage phenotype that induces breast tumor regression and inhibits metastasis [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr B038.
Abstract B052: Peroxisome proliferator-activated receptor delta agonist GW501516 enhances the efficacy of adoptive cell therapy
Authors: Saibil Samuel , St. Paul Michael , Ohahsi Pamela 
Published: November 2016

Abstract
Adoptive cell therapy (ACT) has demonstrated tremendous promise for the treatment of metastatic melanoma. Emerging evidence has indicated that memory CD8+ T cells (Tcm) are superior mediators of ACT than are effector CD8+ T cells (Teff) due to a superior ability to persist in vivo. Underpinning the survival advantage of Tcm is a shift in cellular metabolism away from aerobic glycolysis to instead favor fatty acid oxidation (FAO). This switch to FAO in Tcm requires the expression of carnitine palmitoyl transferase 1a (CPT1a), the rate-limiting enzyme of FAO. Here, we investigated the impact of the peroxisome proliferator-activated receptor (PPAR)δ/β agonist GW501516 (GW), an agent known to boost CPT1a expression and promote FAO in other tissues, on CD8+ T cell metabolism, function and performance in the murine B16-GP33 melanoma model. Via the activation of both PPARα and PPARδ/β we found that GW treatment increased CPT1a expression in activated CD8+ T cells. Using a metabolomics approach, we demonstrated that GW treatment increased the abundance of multiple different acylcarnitines, consistent with enhanced FAO. Unexpectedly, we also found that T cells activated in the presence of GW as well as inflammatory signals, either LPS-matured dendritic cells (DCs) or IL-12, demonstrated enhanced production of interferon-γ. This increase in interferon-γ correlated with increased expression of the transcription factor T-bet. Despite the high T-bet expression, a characteristic of short-lived effector cells, GW-treated CD8+ T cells activated by LPS-treated DCs demonstrated an enhanced ability to persist in vivo. They also demonstrated improved performance in our murine model of ACT. Thus, our data indicates that combined treatment of CD8+ T cells with GW and inflammatory signals can result in in Teff cell with improved ability to produce cytokines but that have also shifted their metabolism to increase FAO akin to Tcm, allowing for enhanced in vivo persistence. These data identify GW501516, and potentially other combined PPARα and PPARδ/β agonists currently being developed, as attractive candidates for further studies and rapid translation into clinical trials of ACT.
Citation Format: Samuel Saibil, Michael St. Paul, Pamela Ohahsi. Peroxisome proliferator-activated receptor delta agonist GW501516 enhances the efficacy of adoptive cell therapy [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr B052.
Abstract B061: CD81: A new target for therapy of B cell lymphoma
Authors: Vences-Catalán Felipe , Kuo Chiung-Chi , Rajapaksa Ranjani , Levy Ronald , Levy Shoshana 
Published: November 2016

Abstract
CD81 belongs to an evolutionary conserved family of proteins named tetraspanins and it was originally described as a B cell target. Tetraspanins associate in multi-molecular complexes in tetraspanin-enriched microdomains (TEMS). Indeed, in B cells CD81 associates with CD19 and CD21, and together function as co-receptor to lower the cell activation threshold initiated by the BCR. Recently, we have demonstrated the importance of CD81 in tumor growth and metastasis of solid tumors1. However, the role of CD81 in B cell malignancies has not been explored. Previously, we have shown that anti-CD81 antibodies had anti-proliferative effects on human B cell lymphoma cell lines in vitro2. Here we tested the therapeutic effect of a mouse anti-human CD81 antibody in vivo against Raji and SUP-B8 B cell lymphomas using a xenograft model in SCID mice. Our studies demonstrated that our anti-CD81 antibody had therapeutic effect comparable to rituximab. Moreover, we found ADCC as one of the mechanism of action. Furthermore, to enhance the anti-CD81-mediated ADCC we engineered chimeric antibodies containing human IgG1ADCC-HIGH and human IgG4 ADCC-LOW Fc constant regions. Indeed, the chimeric human IgG1 anti-CD81 mAb showed a remarkable increase in NK cell-mediated ADCC compared to Rituximab in vitro. These results suggest that CD81 can be a potential therapeutic target on B cell lymphomas by virtue of both its direct cytotoxic effect and as a mediator of ADCC. The Human IgG1 version is being developed as a therapeutic candidate.
1. Vences-Catalan, F., et al. Cancer Res 75, 4517-4526 (2015).
2. Oren, R., Takahashi, S., Doss, C., Levy, R. & Levy, S. Mol Cell Biol 10, 4007-4015 (1990).
Citation Format: Felipe Vences-Catalán, Chiung-Chi Kuo, Ranjani Rajapaksa, Ronald Levy, Shoshana Levy. CD81: A new target for therapy of B cell lymphoma [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr B061.
Abstract B075: Conversion of extracellular signals to programmable genome manipulation via CRISPRouter
Authors: Dingal P C Dave P. , Kipniss Nathan , Gao Yuchen , Qi Lei S. 
Published: November 2016

Abstract
Molecular engineering and synthetic biology approaches have been applied (1) to replace the intracellular domain (NICD) of Notch with synthetic transcription factors1 that bind to genes other than natural Notch targets; (2) to replace human Notch extracellular domain (NECD) with recognition motifs against novel targets (e.g. cognate receptor ligands such as Delta versus single-chain variable fragments raised against a specific antigen). The project aims to interrogate and engineer the fate of a receiver cell to surface-receptor levels of a sender cell by harnessing contact-mediated Notch-Delta signaling and RNA-guided CRISPR genome engineering. We have developed Notch1-Cas9 chimeric receptors that, upon binding Delta ligand to the extra-cellular Notch domain, cleave and release Cas9 to manipulate gene expression of transgenic reporter constructs - and - of endogenous targets in mammalian cells. First, we describe the challenges and solutions in the construction of Notch-Cas9, which led to the discovery of a naturally occurring sequence in bacterial Cas9 that is being co-opted as a nuclear localization signal (NLS) when expressed in mammalian cells. To our knowledge, this is the first reported evidence of a cognate NLS in bacterial Cas9. Second, we expand the toolkit with not only Delta-dependent gene activation, but also with gene editing in two Cas9 orthologs (Streptococcus pyogenes and Staphylococcus aureus Cas9). The toolkit performs well in reporter-gene activation and editing assays. Finally, we demonstrate activation of and phenotypic changes associated with endogenous genes encoding for regulators of the cell-division cycle, CDKN1B and CASP8; and editing of CXCR4 and CD47, receptors relevant to immune trafficking and tolerance. This Notch-Cas9 fusion technology opens up novel biological behaviors in response to cell-cell contact and can be utilized to interrogate or manipulate existing signaling pathways.
Citation Format: P C Dave P. Dingal, Nathan Kipniss, Yuchen Gao, Lei S. Qi. Conversion of extracellular signals to programmable genome manipulation via CRISPRouter [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr B075.
Abstract B087: Revealing antibody repertoires and RAG tracking by linear amplification mediated-high-throughput genome-wide translocation sequencing
Authors: Hu Jiazhi , Lin Sherry G. , Ba Zhaoqing , Du Zhou , Zhang Yu , Alt Frederick W. 
Published: November 2016

Abstract
B lymphocytes generate antibodies to neutralize numerous antigens via RAG endonuclease-initiated V(D)J recombination which joins diverse V and D segments into several J segments. High-throughput genome-wide translocation sequencing (HTGTS) was developed to identify genomic fragments that translocate to a distinct “bait” fragment associated with DNA double-stranded breaks (DSBs). With a single bait primer to linearly amplify across bait-prey junctions for “prey” sequence identification, the linear amplification-mediated HTGTS (LAM-HTGTS) could sensitively detect genome-wide DSBs or translocation in an unbiased way independent of prey sequences. Employing the J-segment-associated coding ends cleaved by RAG as bait fragments, we can capture all the Vs and Ds joined to the bait J segments and thus reveal antibody repertoires in B lymphocyte populations. Such LAM-HTGTS adapted repertoire sequencing (HTGTS-Rep-seq) can easily uncover the productive and non-productive V(D)J joins and complementary determining region 3 (CDR3) information that is important for antigen contact, as well as D usage and RAG “off-targets” that are inviable to previous assays. HTGTS-Rep-seq analysis on the Igk repertoires showed that Vks were widely utilized in spite of their transcription orientations convergent or tandem to those of Jks. However, deletion of the four CTCF-binding elements (CBEs) in the Vk-Jk intervening region greatly increased the usage of Vks proximal to Jks. Strikingly, the usage of Vks transcriptionally convergent to Jks was far more elevated than those in the same transcriptional orientation to Jks, which strongly suggested a role of RAG tracking in mediating increased proximal Vk usage in the absence of Vk-Jk intervening CBEs.
Citation Format: Jiazhi Hu, Sherry G. Lin, Zhaoqing Ba, Zhou Du, Yu Zhang, Frederick W. Alt. Revealing antibody repertoires and RAG tracking by linear amplification mediated-high-throughput genome-wide translocation sequencing [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr B087.
Abstract B094: Targeted expression and molecular profiling assay for tumor microenvironment
Authors: Chenchik Alex , Komarov Andrey , Makhanov Michael , Sastry Sunitha , Frangou Costa 
Published: November 2016

Abstract
New rapid and robust transcriptome-based methods for cellular characterization of the tumor microenvironment and biomarker discovery are required to improve cancer prognosis and treatment. However, challenges with current approaches for the above applications include high sample requirements, poor sensitivity, low dynamic range, and limited throughput. To address these limitations, we have developed a unique approach for targeted transcriptome profiling using validated targeted primers that leverages the sensitivity of multiplex RT-PCR with the throughput of Next-Generation Sequencing (NGS) technology. By combining these methods, just 10-100 ng of total RNA is sufficient to quantify over 5 orders of magnitude variation in gene expression levels. Further, the use of targeted primers enables direct analysis of total RNA isolate and obviates the need for globin depletion from whole blood samples. Finally, using a defined set of amplicons to assess expression levels of all protein-coding genes facilitates and simplifies data analysis and allows more precise sample-to-sample normalization. We will present profiling results that demonstrate how this assay can be used to analyze the level of immune cell infiltration, assess intact and deficient immune mechanisms, and generally elucidate the tumor microenvironment of breast cancer samples.
Citation Format: Alex Chenchik, Andrey Komarov, Michael Makhanov, Sunitha Sastry, Costa Frangou. Targeted expression and molecular profiling assay for tumor microenvironment [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr B094.