bluuebunny/crossref_metadata_2025 huggingface dataset:
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

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

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.
Abstract B119: Characterization of a phosphatidylserine, TAM receptor (Tyro3, Axl, Mertk), PDL1 axis in breast cancer
Authors: Kasikara Canan , Kumar Sushil , Geng Ke , Davre Viral , Empig Cyril , Freimark Bruce , Gray Michael , Schlunegger Kyle , Hutchins Jeff , Kotenko Sergei V. , Birge Raymond B.
Published: November 2016

Tyro3, Axl and Mer (TAMs) are three homologous receptor tyrosine kinases that bind vitamin K-dependent endogenous ligands, Protein S (PROS1) and Gas6, and act as bridging molecules to promote PS-mediated clearance of apoptotic cells (efferocytosis). In recent years, evidence has accumulated that TAMs are overexpressed in a wide array of tumor types, whereby the level of expression correlates with the tumor grade and the emergence of chemo and radio resistance to targeted therapeutics. TAMs have also been implicated as inhibitory receptors on myeloid-derived cells that suppress anti-tumor immunity. In addition to TAM overexpression, externalized PS is also concomitantly up-regulated in the tumor microenvironment, suggesting that PS and PS receptors might act as immune checkpoint inhibitors. To better understand the biology of TAMs, and the specificity of interaction between TAMs and PS, we generated chimeric TAM reporter cell lines comprised of the extracellular domains of each TAM fused to the intracellular domains of the IFNR1, as well as cell lines stably expressing full-length native TAMs. Using these systems, we found that each TAM receptor has a unique pattern of activation by GAS6 and PROS1, as well as unique dependency for PS on apoptotic cells, PS liposomes, and exosomes. Interrogating epithelial cells that express WT TAMs, we also observed that each TAM showed differential capacity for efferocytosis, AKT-mediated chemo-resistance, and their ability to up-regulate the immune checkpoint inhibitory ligand PD-L1 on breast cancer cells. Functionally, TAM-mediated efferocytosis and PD-L1 up-regulation could be partially blocked by PS targeted antibodies 11.31 and bavituximab, suggesting the existence of a PS/PS-R (TAM-receptor)/PD-L1 axis that drives immune escape in solid cancers. These studies demonstrate that, despite their similarities, Tyro3, Axl and Mertk are likely to perform distinct functions in both immune-regulation and the recognition and removal of apoptotic cells. Finally, these studies provide a rationale for combinatorial therapeutics targeting PS, TAM, and PD-L1 as immune checkpoint inhibitors in the treatment of cancer.

Citation Format: Canan Kasikara, Sushil Kumar, Ke Geng, Viral Davre, Cyril Empig, Bruce Freimark, Michael Gray, Kyle Schlunegger, Jeff Hutchins, Sergei V. Kotenko, Raymond B. Birge. Characterization of a phosphatidylserine, TAM receptor (Tyro3, Axl, Mertk), PDL1 axis in breast cancer [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 B119.
Abstract B131: Quantitative modeling of Syk regulation in B cell chronic lymphocytic leukemia
Authors: Frushicheva Maria P.
Published: November 2016

Spleen tyrosine kinases (Syk) are initiators of B cell receptor-mediated (BCR) signaling events, enabling cells to drive chronic lymphocytic leukemia (CLL) progression. However, the mechanistic principles of biochemical signaling pathways that contribute to CLL pathogenesis are poorly understood. This study investigates Syk-mediated molecular pathways of the BCR signaling network using computational modeling. An expression of ζ chain-associated protein kinase of 70 kD (ZAP-70, Syk-family protein kinase) in the BCR signaling pathway is an adverse prognostic marker in CLL. Therefore, we need a better understanding of molecular mechanisms of Syk family protein kinases and effects of their inhibitors in the BCR signaling pathway. In this work, we dissected several computational models of ZAP-70 and SYK regulation to design an integrated mechanistic description of the BCR signaling pathway, and to explain the observed differences in the clinical behaviors of ZAP-70+ / ZAP-70- phenotypes of CLL patients. Specifically, we characterized the effects of different ZAP-70 and SYK expression and phosphorylation levels on the BCR activation threshold, using stochastic simulations of the BCR signaling network. Preliminary findings suggest that ZAP-70 serves as an adaptor protein that facilitates the recruitment of SYK to the BCR complex. We find that the expression of ZAP-70 enhances the BCR signaling, and therefore correlates with a more aggressive disease course. Our results will help in the development of novel therapies targeting Syk functions to regulate B-CLL responses.

Citation Format: Maria P. Frushicheva. Quantitative modeling of Syk regulation in B cell chronic lymphocytic leukemia [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 B131.
Abstract B143: Fc gamma receptor IV mediated depletionof tumor infiltrating regulatory T cells by anti-CTLA4 antibody is promoted byTLR1/2 agonist and hence its efficacy in anti-tumor combination therapy
Authors: Sharma Naveen , Allison James P.
Published: November 2016

Immune checkpoint blockade therapy has been effective but still a significant number of cancer patients either do not respond or become resistant to these therapies. We hypothesized that activating innate immune pathway along with adaptive immune pathway will have a better tumor reduction and survival outcome from immunotherapy treatment. Therefore, we injected TLR1/2 ligand intratumorally in combination with intraperitoneal injection of anti-CTLA4 antibody in a mouse model of melanoma. We show that this combination treatment enhanced anti-tumor immune responses both qualitatively and quantitatively, which is dependent on both CD4 and CD8 T cells. Interestingly, Our results further suggested that increased depletion of regulatory T cells in tumor microenvironment could be one of the mechanisms of enhanced efficacy of combination therapy and this efficacy is dependent upon Fcγ receptor IV expression. This study shows modulation of Fcγ receptor IV expression by a TLR ligand on macrophages to be important in efficacy of anti-CTLA4 antibody in a combination therapy and this mechanism could be harnessed to modulate the final outcome of other antibody-based immunomodulatory therapies.

Citation Format: Naveen Sharma, James P. Allison. Fc gamma receptor IV mediated depletionof tumor infiltrating regulatory T cells by anti-CTLA4 antibody is promoted byTLR1/2 agonist and hence its efficacy in anti-tumor combination 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 B143.
Abstract B145: Small molecule inhibition of cyclic GMP-AMP synthase reduces interferon expression in macrophages from an autoimmune mouse model
Authors: Gao Pu
Published: November 2016

Cyclic GMP-AMP synthase (cGAS) plays a central role in sensing aberrantly localized cytoplasmic dsDNA. Upon binding dsDNA, cGAS synthesizes the cyclic dinucleotide cGAMP, which acts as a second messenger to initiate a signaling pathway for the induction of interferon genes. Given that cytoplasmic DNA can originate from bacteria or virus, cGAS is essential for innate immunity against pathogenic infection. Conversely, cGAS detection of mislocalized nuclear or mitochondrial DNA can lead to hyper-inflammation. Recent studies indicate that cGAS can potentiate autoimmunity in cases of lupus, Aicardi-Goutières syndrome, and other related diseases—suggesting that targeting cGAS activity may be therapeutically beneficial. We report the discovery of a first-in-class generation of cGAS small molecule inhibitor, RU.365, identified in a high-throughput screen. Its structure-activity relationship, and selectivity and potency in cellular assays, was characterized. Moreover, we solved the structure of cGAS in complex with dsDNA and RU.365 or its close structural analog within the active site. Finally, we demonstrate that the inhibitor can reduce constitutive expression of interferonin macrophages derived from a mouse model of autoimmunity. RU.365 will be useful towards understanding the biological roles of cGAS and can serve as a molecular scaffold for development of future autoimmune therapies.

Note: This abstract was not presented at the conference.

Citation Format: Pu Gao. Small molecule inhibition of cyclic GMP-AMP synthase reduces interferon expression in macrophages from an autoimmune mouse model [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 B145.
Abstract IA07: Antitumor T cells: You are what you eat
Authors: Cui Guoliang , Ho Ping-Chih , Amezquita Robert , Kaech Susan
Published: November 2016

Activated T cells engage aerobic glycolysis and anabolic metabolism for growth, proliferation, and effector functions. In contrast, resting memory CD8 T cells depend less on high rates of glycolysis and more on fatty acid oxidation to persist, self-renew, and respond to secondary infection. This talk will discuss the concept of metabolic checkpoints for immune cells in tumors, referring to the idea that the nutrients utilized by different types of T cells can affect both their energetic demands and their functionality. In particular, the metabolic state of exhausted T cells found in tumors has only recently been studied. Our data in mouse models of cancer suggest that a glucose-poor, fatty acid-rich tumor microenvironment limits aerobic glycolysis, but promotes fatty acid uptake in tumor-infiltrating T cells, which suppresses tumoricidal effector functions and increases PD-1 expression. This talk will discuss recent analyses of the signaling pathways that respond to these nutrient alterations in T cells in tumors and how this may relate to new modalities of treatment in combination with checkpoint blockade.

Citation Format: Guoliang Cui, Ping-Chih Ho, Robert Amezquita, Susan Kaech. Antitumor T cells: You are what you eat [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 IA07.
Abstract IA08: Identifying and overcoming metabolic checkpoints to antitumor immunity
Authors: Delgoffe Greg M.
Published: November 2016

It has now become clear that the immune system is not oblivious to cancer initiation and progression, but rather is rendered dysfunctional through a wide array of tolerogenic mechanisms. One of these is the generation of a potently immunosuppressive microenvironment, characterized by altered populations of suppressive cells, secretion of suppressive cytokines, and upregulation of inhibitory ligands. The tumor microenvironment also possesses, due to the deregulated energetics of cancer cells, an altered metabolic profile: hypoxia, acidosis, and a dearth of glucose and other carbon sources. T cell activation and effector function are metabolically demanding, requiring elevated glucose processing and mitochondrial activity to keep up with the bioenergetic demands of proliferation, cytokine secretion, and cytolytic capacity. Thus, we sought to determine whether the metabolism of the tumor microenvironment acts as a barrier to antitumor immunity and resistance mechanism to immunotherapy. We have demonstrated that T cells infiltrating murine and human tumors exhibit a phenotype of metabolic insufficiency, characterized by decreased glucose uptake and a crippling loss of mitochondrial function. We show that this loss of oxidative function is specific to the chronic activation observed in the tumor microenvironment, and occurs rapidly upon entry into the tumor. This loss of mitochondrial activity and mass is due, in part, to the repression of PGC1α, a transcriptional co-activator that programs mitochondrial biogenesis. Further, we have identified chronic Akt activation and consequent Foxo inactivation as a major driver of PGC1α repression. Importantly, metabolic reprogramming of tumor-specific T cells to enforce mitochondrial biogenesis results in dramatically increased antitumor immunity, characterized by robust proliferation, cytokine production, and tumor regression and clearance. Our data support a model in which mitochondrial insufficiency represents a primordial checkpoint inhibiting antitumor immunity in addition to immunologic suppression. Further supporting this notion are additional studies from our laboratory suggesting that selective boosting of T cell mitochondrial activity through mitigation of tumor hypoxia dramatically enhances immunotherapies like checkpoint blockade and adoptive cell therapy. Our data suggest not only that the metabolic nature of the tumor microenvironment may be predictive of a patient's response to various immunotherapies, but that metabolic reprogramming of T cells or the microenvironment itself may be an important means to improve the immunotherapeutic treatment of cancer.

Citation Format: Greg M. Delgoffe. Identifying and overcoming metabolic checkpoints to antitumor immunity [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 IA08.
Abstract IA13: Immune suppressive and immune stimulating monocytes in cancer
Authors: Bronte Vincenzo
Published: November 2016

The tumor environment is enriched in myeloid cells, which can either promote or fight tumor progression. By releasing soluble mediators—such as cytokines, interleukins, metabolites, and growth factors—tumors stimulate a reactive myelopoiesis at both medullary and extra-medullary sites such as the spleen, which alters the steady-state myeloid differentiation, pushing proliferation and expansion of myeloid elements that acquire immunosuppressive and pro-tumoral functions. This process is regulated by the fine balance between pro-survival and anti-apoptotic programs in myeloid cells. The frequency and the function of myeloid-derived suppressor cells (MDSC) in the blood of tumor patients is now regarded as a negative prognostic marker, which correlates with the clinical outcome and response to therapy, including both conventional chemotherapy as well as immunotherapy. Recent data suggest that the monocytic compartment within MDSC (M-MDSC) in tumor-conditioned hosts is extremely plastic. Different pathways activated in M-MDSC, including the metabolism of the semi essential amino acid L-arginine, exert a negative influence on the proliferation and functions of antigen-stimulated T lymphocytes. In fact, the inability of T lymphocytes to recognize and destroy tumor cells can depend on the suppressive milieu shaped by the two enzymes involved in L-arginine metabolism. On the other hand, nitric oxide-producing cells derived from monocytes can support cancer cell destruction and tumor regression. In fact, even though tumor-specific CD8+ T cells are negatively influenced by intra-tumoral L-arginine metabolism, they can license a subset of monocytes to aid tumor rejection, which depends on increased tumor antigen presentation and nitric oxide synthase 2 induction, as the result of the intratumor activation of the CD40-CD40L axis. Understanding the molecular events and cell biology of immune stimulating and immune suppressive monocytes can provide additional targets to modify the tumor environment and enhance cancer immunotherapy.

Citation Format: Vincenzo Bronte. Immune suppressive and immune stimulating monocytes in cancer [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 IA13.
Abstract IA27: Novel natural killer cell targets for cancer immunotherapy
Authors: Smyth Mark J. , Putz Eva , Blake Stephen , Gao Aaron , Guimaraes Fernando , Miles John , Dougall Bill , Teng Michele WL , Huntington Nicholas D.
Published: November 2016

Avoiding immune destruction is a hallmark of cancer. Natural killer (NK) cells contribute to tumor immune surveillance and protection from tumor metastasis. The cell surface Ig superfamily molecule CD96 has recently been shown as a negative regulator of mouse NK cell activity, and we have demonstrated that blocking CD96 with a monoclonal antibody inhibited experimental metastases in multiple tumor models. The anti-metastatic activity of anti-CD96 was dependent on NK cells, CD226 (DNAM-1) and IFN-γ, but independent of activating Fc receptors. Anti-CD96 was more effective in combination with anti-CTLA4, anti-PD1, anti-TIGIT, or doxorubicin chemotherapy. Development of additional anti-mouse and anti-human CD96 mAbs will be discussed. Overall these data demonstrate that blocking CD96 is a new and complementary immunotherapeutic strategy to reduce tumor metastases. The activity of NK cells is also governed by the cytokine IL-15, foreign and self-ligands, and a variety of immunosuppressive factors. We have identified CIS (Cytokine-inducible SH2-containing protein; Cish gene) as the critical negative regulator of IL-15 signalling in NK cells and shown the critical intrinsic role of CIS in suppressing NK cell control of tumor initiation and metastasis. The combination of Cish-deficiency and relevant immunotherapies, such as immune checkpoint blockade antibodies, IL-2, and type I interferon revealed further improved control of metastasis. The data clearly indicates that targeting CIS promotes NK cell antitumor functions and CIS holds great promise as a novel target in NK cell immunotherapy. The relative importance of mechanisms that tumors adopt to evade NK cells remains ill defined. Using mice with NK cells that either constitutively, or are unable to, signal via the TGF-β receptor, revealed that TGF-β signaling induces the transdifferentiation of conventional NK (cNK) cells into ILC1. Functional analysis revealed that host protection from tumor initiation and metastasis is mediated by cNK cells, whereas tumor resident ILC1 and ILC1-like cells appear to offer no host protection. These data indicate the plasticity of cNK cells under pathophysiological conditions and a potential new escape mechanism for tumors from innate immunity.

Citation Format: Mark J. Smyth, Eva Putz, Stephen Blake, Aaron Gao, Fernando Guimaraes, John Miles, Bill Dougall, Michele WL Teng, Nicholas D. Huntington. Novel natural killer cell targets for cancer immunotherapy [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 IA27.
Abstract IA29: Manipulating inflammation to raise cancer immunogenicity
Authors: Zelenay Santiago
Published: November 2016

Cyclooxygenase (COX)-2, one of the rate-limiting enzymes for production of prostanoids, is commonly upregulated in numerous cancers, and together with prostaglandin E2 (PGE2), has been implicated in several aspects of malignant growth including proliferation, angiogenesis, and invasion. We found that COX-dependent production of PGE2 by melanoma cells dominantly and primarily enables progressive tumor growth via immune escape. Melanoma cells, unable to produce PGE2 by means of genetic ablation of COX or prostaglandin E synthase, formed spontaneously regressing tumors in immune-competent mice but progressive tumors in immune-deficient hosts. By contrast, their PGE2-competent parental lines grew progressively and irrespectively of the presence of a fully competent immune system, demonstrating an essential role for PGE2 in immune evasion. Importantly, PGE2-dependent immune escape was crucial not only in melanoma but also in colorectal and breast cancer models, arguing that this is a general mechanism for subversion of anticancer immunity. COX deficiency in cancer cells was associated with a marked shift in the inflammatory signature at the tumor site characterized by lower expression of cancer-promoting factors and concomitant increase in several mediators typically associated with antitumor immunity. These drastic changes in the inflammatory nature of the tumor microenvironment due to reduced PGE2 levels suggested that COX inhibitors could be used to potentiate immune-dependent control. Indeed, we found that combining COX-inhibitors, such as aspirin, with anti-PD-1 blockade synergistically promoted growth control of tumors poorly responsive to either monotherapy. Finally, analysis of publicly available human melanoma datasets revealed a remarkable conservation of the COX-driven mouse inflammatory signature suggesting that COX activity might be a driver of immune suppression across species.

Citation Format: Santiago Zelenay. Manipulating inflammation to raise cancer immunogenicity [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 IA29.
Abstract IA30: Technologies for personalizing cancer immunotherapies
Authors: Heath James R. , Peng Songming , Hsu Alice , Esswein Shannon , Heath John , Noh Won Jun , Zaretsky Jesse , Ribas Toni
Published: November 2016

At the heart of most cancer immunotherapies are the specific interactions between the principal cancer cell killers, T cells, and antigens presented by the tumor cells. Those interactions may be exposed through the use of checkpoint inhibitors, or they can be amplified through the use of engineered cell-based therapies. A class of antigens that has emerged as being particularly important are neoantigens, which are small fragments of mutated proteins that contain the mutation, and, for CD8+ T cell recognition, are presented by MHC Class I. In principle, neoantigens that draw T cells into a tumor can comprise personalized vaccines, and the T cell receptors (TCRs) that recognize those neoantigens can be engineered as personalized cellular therapies. However, identifying which neoantigens are drawing which T cells into the tumor and promoting cell killing has been a challenge. In this talk I will describe an approach, called nanoparticle-barcoded nucleic acid cell sorting (NP-barcoded NACS), designed for this purpose. The approach begins with a list of putative neoantigens generated from in silico analysis of the tumor genome and transcriptome. Those neoantigens are prepared as a NP-barcoded NACS library so that each neoantigen is paired with a DNA barcode. The entire library is then used to precipitate neoantigen specific CD8+ T cell populations from the patient's tumor or blood. Each cell is then individually decoded using sequential fluorescent reads to identify its cognate antigen, and the T cells may be further separated for single cell TCR sequencing. We have used this method to analyze tumor materials and blood from melanoma cancer patients responding to anti-PD-1 therapy, mouse models of immunotherapy, and tumor materials collected from various other cancer patients. The method, which is shown to be at least an order of magnitude more sensitive than alternative approaches, requires very little material, and so may be used for the analysis of non-expanded tumor infiltrating lymphocytes (TILs) or peripheral blood mononuclear cells (PBMCs). I will discuss some of the analytical details of this method, and prospects for harnessing it to develop personalized immunotherapies. I will also provide results of kinetic studies, via blood analysis, that appear to provide insights into patient responses to checkpoint inhibitor therapy.

Citation Format: James R. Heath, Songming Peng, Alice Hsu, Shannon Esswein, John Heath, Won Jun Noh, Jesse Zaretsky, Toni Ribas. Technologies for personalizing cancer immunotherapies [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 IA30.
Characterization of Human AB Serum for Mesenchymal Stromal Cell Expansion
Authors: dos Santos Vanessa Tieko Marques , Mizukami Amanda , Orellana Maristela Delgado , Caruso Samia Rigotto , da Silva Fernanda Borges , Traina Fabiola , de Lima Prata Karen , Covas Dimas Tadeu , Swiech Kamilla
Published: November 2016

Background: So far, using human blood-derived components appears to be the most efficient and safest approach available for mesenchymal stromal cell (MSC) expansion. In this paper, we report on the characterization of human AB serum (AB HS) produced by using different plasma sources, and its use as an alternative supplement to MSC expansion. Methods: Two plasma sources were used for AB HS production: plasma removed from whole blood after 24 h of collection (PC > 24 h) and plasma, cryoprecipitate reduced (PCryoR). The biochemical profile and quality of the produced AB HS batches were analyzed and their ability to support MSC cell growth after different storage times (0, 3, 6, 9 and 12 months) was evaluated. Results: The two plasma sources used showed similar characteristics regarding biochemical constituents and quality parameters and were effective in promoting MSC growth. MSCs cultured in medium supplemented with 10% AB HS presented similar doubling times and cumulative population doublings when compared to the 10% fetal bovine serum(FBS)-supplemented culture while maintaining immunophenotype, functional features, and cytogenetic profile. Conclusion: Overall, the results indicate that AB HS is an efficient FBS substitute and can be used for at least 12 months after production without impairing cell proliferation and quality.
Genomic Microarray in Fetuses with Early Growth Restriction: A Multicenter Study
Authors: Borrell Antoni , Grande Maribel , Meler Eva , Sabrià Joan , Mazarico Edurne , Muñoz Anna , Rodriguez-Revenga Laia , Badenas Cèlia , Figueras Francesc
Published: November 2016

Background: Little information is available about the risk of microdeletion and microduplication syndromes in fetal growth restriction (FGR) with a normal karyotype. Objective: To assess the incremental yield of genomic microarray over conventional karyotyping in fetuses with early growth restriction. Study Design: Genomic microarray was prospectively performed in fetuses with early growth restriction defined as a fetal weight below the 3rd percentile estimated before 32 weeks of pregnancy, and a normal quantitative fluorescent polymerase chain reaction result. The incremental yield of genomic microarray was defined by the rate of fetuses presenting with a pathogenic copy number variant below 10 Mb. Results: Among 133 fetuses with early FGR, a 6.8% (95% CI: 2.5-11.0) incremental yield of genomic microarray over karyotyping was observed. This incremental yield was 4.8% (95% CI: 0.2-9.3) in isolated FGR, 10% (95% CI: 0-20.7) in FGR with nonstructural anomalies, and 10.5% (95% CI: 0-24.3) in FGR with structural anomalies. Conclusion: Our multicenter study reveals that 6.8% of fetuses with early growth restriction present with submicroscopic anomalies after common aneuploidies were excluded. Even when FGR is observed as an isolated finding, genomic microarray analysis should be considered after or instead of karyotyping, due to its 4.8% incremental yield.
Urinary Periostin Excretion Predicts Renal Outcome in IgA Nephropathy
Authors: Hwang Jin Ho , Lee Jung Pyo , Kim Clara Tammy , Yang Seung Hee , Kim Jin Hyuk , An Jung Nam , Moon Kyung Chul , Lee Hajeong , Oh Yun Kyu , Joo Kwon Wook , Kim Dong Ki , Kim Yon Su , Lim Chun Soo
Published: November 2016

Background: Periostin is a matricellular protein and plays a vital role in tissue regeneration, fibrosis and wound healing. However, data about its significance in nephrology are limited. We investigated the correlation between urinary periostin excretion and its clinical significance including renal histologic findings and prognosis in IgA nephropathy (IgAN). Methods: Of 399 patients from a glomerulonephritis cohort recruited between January 2009 and December 2014, 314 were enrolled. Serum and urine periostin (uPOSTN) were measured using enzyme-linked immunosorbent assay. We divided the patients into 3 groups by uPOSTN/creatinine (uPOSTN/Cr): group 1 (undetectable), group 2 (lower than the median) and group 3 (higher than the median). Results: The uPOSTN level was correlated with pathologic classifications and both initial and final IDMS-MDRD estimated glomerular filtration rates (eGFRs; p < 0.001). Histologically, group 3 patients were correlated with severe interstitial fibrosis/tubular atrophy (p = 0.004), interstitial inflammation (p = 0.007), hyaline arteriolosclerosis (p = 0.001) and glomerular sclerosis (p < 0.001). A higher initial uPOSTN/Cr level was associated with a greater decline in eGFR during follow-up (p = 0.043 when initial eGFR ≥60; p = 0.025 when eGFR <60 ml/min/1.73 m2), and the renal outcomes with end-stage renal disease (ESRD; p = 0.003), ESRD and/or eGFR decrease of >30% (p = 0.033) and ESRD and/or eGFR decrease of >50% (p = 0.046) occurred significantly more in group 3. In multivariate analysis, uPOSTN group 3 (hazards ratio 2.839, 95% CI 1.013-7.957; p = 0.047) was independently associated with ESRD in IgAN patients. Conclusion: uPOSTN/Cr value at initial diagnosis correlated with renal fibrosis and predicted the renal outcomes in patients with IgAN. It could be a promising urinary biomarker for renal fibrosis.
Contribuições para um bestiário feminino em Plauto
Authors: Rocha Carol Martins da
Published: November 2016

A disputa entre homens e mulheres é um dos motivos cômicos típicos da palliata, gênero a que pertencem as 21 comédias do poeta romano Tito Mácio Plauto (III–II a.C.). Assim, muitos dos enredos dessas peças giram em torno de uma verdadeira guerra, sobretudo, entre maridos e esposas. Buscando perceber de que maneira essa disputa entre os sexos é caracterizada, observamos exemplos de invectivas tanto de personagens femininos contra masculinos, como o contrário, nas peças Báquides, Cásina, Menecmos e O Mercador. Nosso interesse principal diz respeito a uma possível construção de um repertório “bélico”, envolvendo o uso recorrente de metáforas animais para caracterizar as mulheres.
Adivinhação e poder nas Coéforas de Ésquilo
Authors: Paoli Beatriz De
Published: November 2016

O sonho profético de Clitemnestra e o oráculo de Apolo desempenham, inegavelmente, um papel de grande importância na trama das Coéforas de Ésquilo. Neste artigo, vê-se como estes e outros sinais igualmente pertencentes ao âmbito da arte divinatória, além de contribuírem significativamente para o desenvolvimento da ação trágica, permitem que se vislumbrem os desígnios dos deuses e, dessa forma, mais bem se compreendam os pontos de vista humano e divino, bem como as relações que se estabelecem – particularmente as que dizem respeito ao exercício do poder – entre as personagens mortais e divinas desta segunda peça da trilogia esquiliana.
Hipólito e Teseu enredados nas malhas da decisão indiscutível
Authors: Agostini Cristina De Souza
Published: November 2016

O presente artigo propõe a discussão acerca das relações de poder estabelecidas pelos personagens Hipólito e Teseu, da tragédia Hipólito, de Eurípides. Nesse sentido, tendo como pano de fundo as considerações de Max Weber e Émile Benveniste para o uso do termo poder, pretendo demonstrar de que modo, no drama, a sanção da morte de Hipólito deve, necessariamente, ficar a cargo de Teseu, uma vez que este é o único personagem que, no contexto político da peça, detém legitimamente o poder sobre todos aqueles que estão sob seu domínio. Portanto, se, de um lado, no prólogo, Afrodite apresenta aos espectadores todo o desenvolvimento da tragédia, demonstrando sua atuação direta na ruína da família de Hipólito, de outro lado, Eurípides, de modo verossimilhante, vale-se da legitimidade do poder real de Teseu para selar com o veredito político humano a vontade da deusa Cípris.
Plauto e o triunfo da tragédia
Authors: Costa Lilian Nunes da
Published: November 2016

A presença de elementos variados da tragédia, da épica e mesmo da poesia lírica na comédia de Tito Mácio Plauto (c. 254–184 a.C.) tem sido reconhecida por estudiosos como, respectivamente, Christenson (2000), Oniga (1985) e Traill (2005). Contudo, tal mescla de gêneros poéticos presente no corpus da obra plautina está longe de representá-los em uma convivência pacífica. Ao contrário: em vários momentos a evocação de outros gêneros poéticos parece apontar para uma atmosfera de emulação entre o gênero cômico e aqueles. Nossas pesquisas vêm apontando para o predomínio de alusões à tragédia (ou intermediadas por ela). Mas, como gostaríamos de frisar, não se trata de uma mera questão de quantidade de referências: algumas vezes, essa disputa (e a hierarquia tradicional) é mesmo tematizada, como em Anfitrião, Os cativos, Truculento. Na presente exposição, procuraremos observar mais de perto, em passagens de Plauto, o suposto triunfo que a tragédia teria sobre a comédia, a fim de apurar em que medida isso é verificável em algumas de suas peças.
Phosphodiesterase 2 Protects Against Catecholamine-Induced Arrhythmia and Preserves Contractile Function After Myocardial Infarction
Authors: Vettel Christiane , Lindner Marta , Dewenter Matthias , Lorenz Kristina , Schanbacher Constanze , Riedel Merle , Lämmle Simon , Meinecke Simone , Mason Fleur E. , Sossalla Samuel , Geerts Andreas , Hoffmann Michael , Wunder Frank , Brunner Fabian J. , Wieland Thomas , Mehel Hind , Karam Sarah , Lechêne Patrick , Leroy Jérôme , Vandecasteele Grégoire , Wagner Michael , Fischmeister Rodolphe , El-Armouche Ali
Published: November 2016

Rationale:

Phosphodiesterase 2 is a dual substrate esterase, which has the unique property to be stimulated by cGMP, but primarily hydrolyzes cAMP. Myocardial phosphodiesterase 2 is upregulated in human heart failure, but its role in the heart is unknown.

Objective:

To explore the role of phosphodiesterase 2 in cardiac function, propensity to arrhythmia, and myocardial infarction.

Methods and Results:

Pharmacological inhibition of phosphodiesterase 2 (BAY 60–7550, BAY) led to a significant positive chronotropic effect on top of maximal β-adrenoceptor activation in healthy mice. Under pathological conditions induced by chronic catecholamine infusions, BAY reversed both the attenuated β-adrenoceptor–mediated inotropy and chronotropy. Conversely, ECG telemetry in heart-specific phosphodiesterase 2-transgenic (TG) mice showed a marked reduction in resting and in maximal heart rate, whereas cardiac output was completely preserved because of greater cardiac contraction. This well-tolerated phenotype persisted in elderly TG with no indications of cardiac pathology or premature death. During arrhythmia provocation induced by catecholamine injections, TG animals were resistant to triggered ventricular arrhythmias. Accordingly, Ca
2+
-spark analysis in isolated TG cardiomyocytes revealed remarkably reduced Ca
2+
leakage and lower basal phosphorylation levels of Ca
2+
-cycling proteins including ryanodine receptor type 2. Moreover, TG demonstrated improved cardiac function after myocardial infarction.

Conclusions:

Endogenous phosphodiesterase 2 contributes to heart rate regulation. Greater phosphodiesterase 2 abundance protects against arrhythmias and improves contraction force after severe ischemic insult. Activating myocardial phosphodiesterase 2 may, thus, represent a novel intracellular antiadrenergic therapeutic strategy protecting the heart from arrhythmia and contractile dysfunction.
Blood Pressure–Lowering Effect of Orally Ingested Nitrite Is Abolished by a Proton Pump Inhibitor
Authors: Montenegro Marcelo F. , Sundqvist Michaela L. , Larsen Filip J. , Zhuge Zhengbing , Carlström Mattias , Weitzberg Eddie , Lundberg Jon O.
Published: November 2016

Inorganic nitrate and nitrite from dietary and endogenous sources are metabolized to NO and other bioactive nitrogen oxides that affect blood pressure. The mechanisms for nitrite bioactivation are unclear, but recent studies in rodents suggest that gastric acidity may influence the systemic effects of this anion. In a randomized, double-blind, placebo-controlled crossover study, we tested the effects of a proton pump inhibitor on the acute cardiovascular effects of nitrite. Fifteen healthy nonsmoking, normotensive subjects, aged 19 to 39 years, were pretreated with placebo or esomeprazole (3×40 mg) before ingesting sodium nitrite (0.3 mg kg
−1
), followed by blood pressure monitoring. Nitrite reduced systolic blood pressure by a maximum of 6±1.3 mm Hg when taken after placebo, whereas pretreatment with esomeprazole blunted this effect. Peak plasma nitrite, nitrate, and nitroso species levels after nitrite ingestion were similar in both interventions. In 8 healthy volunteers, we then infused increasing doses of sodium nitrite (1, 10, and 30 nmol kg
−1
min
−1
) intravenously. Interestingly, although plasma nitrite peaked at similar levels as with orally ingested nitrite (≈1.8 µmol/L), no changes in blood pressure were observed. In rodents, esomeprazole did not affect the blood pressure response to the NO donor, DEA NONOate, or vascular relaxation to nitroprusside and acetylcholine, demonstrating an intact downstream NO-signaling pathway. We conclude that the acute blood pressure–lowering effect of nitrite requires an acidic gastric environment. Future studies will reveal if the cardiovascular complications associated with the use of proton pump inhibitors are linked to interference with the nitrate–nitrite–NO pathway.
Genome-Wide and Gene-Based Meta-Analyses Identify Novel Loci Influencing Blood Pressure Response to Hydrochlorothiazide
Authors: Salvi Erika , Wang Zhiying , Rizzi Federica , Gong Yan , McDonough Caitrin W. , Padmanabhan Sandosh , Hiltunen Timo P. , Lanzani Chiara , Zaninello Roberta , Chittani Martina , Bailey Kent R. , Sarin Antti-Pekka , Barcella Matteo , Melander Olle , Chapman Arlene B. , Manunta Paolo , Kontula Kimmo K. , Glorioso Nicola , Cusi Daniele , Dominiczak Anna F. , Johnson Julie A. , Barlassina Cristina , Boerwinkle Eric , Cooper-DeHoff Rhonda M. , Turner Stephen T.
Published: November 2016

This study aimed to identify novel loci influencing the antihypertensive response to hydrochlorothiazide monotherapy. A genome-wide meta-analysis of blood pressure (BP) response to hydrochlorothiazide was performed in 1739 white hypertensives from 6 clinical trials within the International Consortium for Antihypertensive Pharmacogenomics Studies, making it the largest study to date of its kind. No signals reached genome-wide significance (
P
<5×10
−
8
), and the suggestive regions (
P
<10
−5
) were cross-validated in 2 black cohorts treated with hydrochlorothiazide. In addition, a gene-based analysis was performed on candidate genes with previous evidence of involvement in diuretic response, in BP regulation, or in hypertension susceptibility. Using the genome-wide meta-analysis approach, with validation in blacks, we identified 2 suggestive regulatory regions linked to gap junction protein α1 gene (
GJA1
) and forkhead box A1 gene (
FOXA1
), relevant for cardiovascular and kidney function. With the gene-based approach, we identified hydroxy-delta-5-steroid dehydrogenase, 3 β- and steroid δ-isomerase 1 gene (
HSD3B1
) as significantly associated with BP response (
P
<2.28×10

−
4

).
HSD3B1
encodes the 3β-hydroxysteroid dehydrogenase enzyme and plays a crucial role in the biosynthesis of aldosterone and endogenous ouabain. By amassing all of the available pharmacogenomic studies of BP response to hydrochlorothiazide, and using 2 different analytic approaches, we identified 3 novel loci influencing BP response to hydrochlorothiazide. The gene-based analysis, never before applied to pharmacogenomics of antihypertensive drugs to our knowledge, provided a powerful strategy to identify a locus of interest, which was not identified in the genome-wide meta-analysis because of high allelic heterogeneity. These data pave the way for future investigations on new pathways and drug targets to enhance the current understanding of personalized antihypertensive treatment.
Hypertensive Disorders of Pregnancy and Offspring Cardiac Structure and Function in Adolescence
Authors: Timpka Simon , Macdonald‐Wallis Corrie , Hughes Alun D. , Chaturvedi Nishi , Franks Paul W. , Lawlor Debbie A. , Fraser Abigail
Published: November 2016

Background

Fetal exposure to preeclampsia is associated with higher blood pressure and later risk of stroke. We aimed to investigate the associations of maternal preeclampsia, gestational hypertension, and maternal blood pressure change in pregnancy with offspring cardiac structure and function in adolescence.

Methods and Results

Using data from a prospective birth cohort study, we included offspring who underwent echocardiography (mean age, 17.7 years;
SD
, 0.3; N=1592). We examined whether hypertensive disorders of pregnancy were associated with offspring cardiac structure and systolic/diastolic function using linear regression. Using multilevel linear spline models (measurement occasions within women), we also investigated whether rate of maternal systolic/diastolic blood pressure change during pregnancy (weeks 8–18, 18–30, 30–36, and 36 or more) were associated with offspring outcomes. Main models were typically adjusted for maternal age, offspring age and sex, prepregnancy body mass index, parity, glycosuria/diabetes mellitus, education, and maternal smoking. Exposure to maternal preeclampsia (0.025; 95% CI, 0.008–0.043) and gestational hypertension (0.010; 0.002–0.017) were associated with greater relative wall thickness. Furthermore, preeclampsia was also associated with a smaller left ventricular end‐diastolic volume (−9.0 mL; −15 to −3.1). No associations were found between hypertensive disorders of pregnancy and offspring cardiac function. Positive rate of maternal systolic blood pressure change during weeks 8 to 18 was associated with greater offspring left ventricular end‐diastolic volume, left ventricular mass indexed to height
2.7
, and E/A.

Conclusions

Adolescent offspring exposed to maternal preeclampsia had greater relative wall thickness and reduced left ventricular end‐diastolic volume, which could be early signs of concentric remodeling and affect future cardiac function as well as risk of cardiovascular disease.