bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract B79: Proinflammatory cytokine profile of syngeneic models
Authors: Dubauskaite Jolanta , Bradshaw Laura , Merlino Ashley , Rong Xianhui , Gee Connie , Dranoff Glenn , Pinzon-Ortiz Maria 
Published: June 2021

Abstract
Cytokines are central immunoregulators that play key roles in both the upregulation (initiation) and downregulation (turning off) immune responses. They are involved in the control of proliferation, cell survival, differentiation, immune cell activation, cell migration and death. In tumors, cytokines can be expressed by immune cells, stromal cells (fibroblast and endothelial cells), and tumor cells. Depending on the inflammatory state of the tumor microenvironment, they can modulate either an antitumoral response or help to induce cell transformation and malignancy. There is ample literature available for cytokine profiles of patient tumor and serum samples but few reports from mouse syngeneic models of cancer. Here, we describe our studies evaluating the cytokine profiles (systemic and intratumoral) of common syngeneic tumor models. We generated the profiles of proinflammatory cytokines (IL-1 beta, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12 p70, interferon-gamma, CXCL-1 and tumor necrosis factor-alpha) for the most commonly used syngeneic models in immuno-oncology (MC38, B16F10, CT26, Colon26, Renca and 4T1). Data from naive mice and cell lines help to understand any changes to these cytokines in tumor-bearing animals naive to treatment. Additionally, we have treated MC38 tumor-bearing animals with two anti-mouse PD-1 clones and evaluated the cytokine responses at different time-points post treatment. We observed localized changes to the levels of some cytokines in the tumor microenvironment, followed by systemic cytokine alterations. The pretreatment cytokine profiles may help us with model selection to investigate immune and inflammatory modulators. In addition, understanding the cytokine responses during and after treatment can also provide insight into the mechanisms of action of immunomodulatory agents and provide hypotheses for combinations. Together, these findings could provide early information about immune infiltrates after treatment, and in the future could be of use as possible biomarkers of response or safety.
Citation Format: Jolanta Dubauskaite, Laura Bradshaw, Ashley Merlino, Xianhui Rong, Connie Gee, Glenn Dranoff, Maria Pinzon-Ortiz. Proinflammatory cytokine profile of syngeneic models [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr B79.
Abstract B83: RNA-nanoparticles for immunotherapy-resistant head and neck squamous cell carcinoma
Authors: Silver Natalie L. , Garg Rekha R. , Fredenberg Kristianna M. , Mendez Hector R. , Mitchell Duane A. , Sayour Elias J. 
Published: June 2021

Abstract
Introduction: Immunotherapy is promising for patients with treatment-refractory head and neck squamous cell carcinoma (HNSCC). However, &gt;80% of patients do not respond to checkpoint inhibitors, suggesting that the highly immunosuppressive tumor immune microenvironment of HNSCC may play a role in treatment resistance. Our group has developed a novel treatment platform utilizing clinically translatable RNA-nanoparticles (NPs) to remodel the systemic and intratumoral HNSCC microenvironment in favor of an activated immunologic milieu.
Objective: The purpose of this study is to examine the feasibility, safety and immunologic activity of systemic/intratumoral administration of RNA NPs for HNSCC.
Materials and Methods: RNA-NPs were prepared by complexing negatively charged mRNA with nanoliposomes. These complexes were injected into naive and B16 or MOC1/MOC2 (murine oral cavity squamous cell carcinoma cell lines) tumor bearing C57Bl/6 mice. In vitro transfection experiments were performed in human HNSCC cell lines.
Results: Human HNSCC cell lines were efficiently transfected with RNA NPs in vitro. MOC1/MOC2 tumors were efficiently transfected with RNA NPs in vivo. RNA-NPs activate dendritic cells (DCs) in the peripheral and intratumoral microenviroment of naive and B16 tumor bearing mice, and mediate antitumor efficacy in a preclinical in vivo murine model for HNSCC. RNA-NPs mediate their effects through interferon-alpha released from plasmacytoid DCs. We have successfully scaled up production of RNA-NPs for translation into human clinical trials and demonstrated safety of RNA-NPs in acute/chronic murine toxicity studies based on CBCs, electrolytes, organ function tests, and end-organ H&E histology.
Conclusions: RNA-NPs safely reprogram the peripheral and intratumoral immune microenvironments, unlocking antitumor activity. We have successfully generated RNA-NPs, providing a renewable resource that can be made readily available for all HNSCC patients. FDA-IND enabling studies are currently under way in preparation for first-in-human trials for checkpoint refractory HNSCC.
Citation Format: Natalie L. Silver, Rekha R. Garg, Kristianna M. Fredenberg, Hector R. Mendez, Duane A. Mitchell, Elias J. Sayour. RNA-nanoparticles for immunotherapy-resistant head and neck squamous cell carcinoma [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr B83.
Abstract B84: Decipher the role of IL-33 as an activator of NK cells’ antitumor activity
Authors: Eberhardt Anais , Blanc Elena , Lardenois Emilie , Renaudineau Sarah , Herbulot Jennifer , Dumont Benoit , Durand Isabelle , Charrier Emilie , Schmitt Pauline , Girard Jean-Philippe , Marçais Antoine , Walzer Thierry , Caux Christophe , Bendriss-Vermare Nathalie 
Published: June 2021

Abstract
Interleukin (IL)-33 is an alarmin belonging to the IL-1 family. Through its receptor ST2, IL-33 promotes both type 1 and type 2 immune responses, depending on the type of responding cells and the microenvironment in damaged tissues. Consistently, paradoxical roles of IL-33 have been reported in the context of cancer. These observations prompted us to investigate i) the expression of IL-33/ST2 in breast tumors and ii) the role of IL-33/ST2 in NK cell activation in physiologic and tumor contexts. Using immunohistochemistry (IHC) and transcriptomic analyses, we observed that IL-33 is expressed at higher levels in ductal carcinoma in situ (DCIS) compared to invasive breast cancer (IBC), especially in the stroma and in luminal BC subtype. In situ, IL-33 was mainly detected in endothelial cells and at lower extent in scattered cells within the stroma of breast tumors. Furthermore, we unraveled a new pathway for NK cells’ activation where IL-12 upregulates ST2 on human NK cells, which in turn become responsive to IL-33 by secreting high levels of IFN-γ and increasing their cytotoxic activity. This effect was specific to a subset of human CD56dim NK cells (20%) and was dependent on STAT-4 phosphorylation. In mice, we also observed a strong activation of spleen NK cell by IL-33 in combination with IL-12 in vitro. Finally, using IL-33ko mice, preliminary results show a contribution of endogenous IL-33 in the prevention of experimental lung metastasis development following B16 iv injection in therapeutic settings using exogenous administration of IL-12. Following up on these results, our aim is now to i) better characterize IL-33-responsive NK cells subpopulation in blood, ii) understand why human CD56bright NK cells do not upregulate ST2 despite response to IL-12 by phosphorylating STAT-4, iii) characterize NK cells’ response to IL-33 in human breast tumors, iv) evaluate ST2 expression by immune cells infiltrating breast and ovarian tumors, and v) confirm the activating role of IL-33 on NK cells’ biology in tumor models in vivo. All together, our observations are in favor of an NK-mediated antitumor role of IL-33 that we are currently pursuing as a potential novel therapeutic strategy in cancer.
Citation Format: Anais Eberhardt, Elena Blanc, Emilie Lardenois, Sarah Renaudineau, Jennifer Herbulot, Benoit Dumont, Isabelle Durand, Emilie Charrier, Pauline Schmitt, Jean-Philippe Girard, Antoine Marçais, Thierry Walzer, Christophe Caux, Nathalie Bendriss-Vermare. Decipher the role of IL-33 as an activator of NK cells’ antitumor activity [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr B84.
Abstract B89: Determining the underlying protective mechanisms of bivalent Marek’s disease vaccine to prevent tumor induction
Authors: Umthong Supawadee , Cheng Hans , Dunn John 
Published: June 2021

Abstract
Marek’s disease (MD) is a lymphoproliferative disease in chicken and the most frequently diagnosed cancer in the animal kingdom. The causative pathogen is the ubiquitous Marek’s disease virus (MDV), a highly oncogenic alphaherpesvirus. Since the 1970s, poultry flocks have been controlled by widespread vaccination (e.g., 1 million birds per hour in the US alone); MD vaccines were revolutionary because they were the first to prevent tumors. Unfortunately, MD vaccines do not prevent viral infection or spread, which has likely promoted the repeated evolution of MDV field strains to higher virulence. Hence, in order to develop more effective MD vaccines, especially those that might prevent horizontal transmission, it is important to gain knowledge on the underlying mechanisms on tumor prevention and viral replication. In addition, chickens serve as a natural model of the disease to study viral oncology and cancer vaccines. The current study focuses on understanding how the commonly used MD vaccines SB-1 and HVT work better in combination (88% protection) compared to either SB-1 or HVT vaccine alone (44% and 24% protection, respectively). We find that bivalent SB-1+HVT vaccine is able to suppress MDV replication in peripheral blood mononuclear cells (PBMCs) until the end of the trial (8 weeks) while either monovalent vaccine failed to control MDV viremia beyond 2 weeks post-challenge. SB-1+HVT also shows the highest efficacy to inhibit tumor formation compared to either monovalent vaccine. To gain insight of how SB-1+HVT can inhibit tumors, we determined the replication of SB-1 and HVT vaccine and find that replication of SB-1 and HVT is different with respect to time after vaccination and tissue tropism. Specifically, SB-1 replicates increasingly in spleen from 1 day post challenge (DPC) to 14 DPC, contrasting to the replication of HVT, which is found only in bursa at 1 DPC. The differences in replication pattern of SB-1 and HVT vaccine may promote different sets of immune cells corresponding to immune protection. Additionally, SB-1+HVT shows the highest immune activation observed by the highest percent CD8 T-cell response. Further experiments are under way to determine whether the induced CD8 response results in antiviral effect at the early stage of infection or antitumor activity as well as identifying associated cytokines induced in the protection provided by SB-1+HVT vaccination.
Citation Format: Supawadee Umthong, Hans Cheng, John Dunn. Determining the underlying protective mechanisms of bivalent Marek’s disease vaccine to prevent tumor induction [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr B89.
Abstract IA07: Hitting the target: T-cell receptors, cross-reactivity, and tumors
Authors: Thomas Paul G. 
Published: June 2021

Abstract
Cancer arises due to an accumulation of genetic alterations, which can lead to the production of mutant proteins that are not expressed by normal cells. Some of these mutant proteins can be processed into short peptides and presented on the cell surface by major histocompatibility complex molecules as neoepitopes and CD8+ T cells can effectively recognize and mount responses against these neoepitopes across various cancer types. However, tumor-specific lymphocytes are generally characterized by an exhausted phenotype, which can in some circumstances be reversed with checkpoint blockade therapies that restore antitumor activity, with those cancer types and individual tumors that bear relatively high mutation burdens showing more robust responses. Using predictive algorithms to identify putative endogenous antitumor T-cell responses across solid tumor types has resulted in an average 2% of predicted neoepitopes associated with a detectable response. This has led to the assumption that low mutation burden tumors, which include many pediatric tumors, are poorly immunogenic. Here, we report that pediatric patients with acute lymphocytic leukemia (ALL) possess broad pools of tumor-associated neoepitope-specific CD8+ T cells, responding to 86% of tested neoantigens and recognizing 68% of the tested neoepitopes. We characterized phenotypic and transcriptional profiles of these epitope-specific antitumor CD8+ T cells at the single-cell level and found a heterogeneous population that included many highly functional effector cells. Moreover, we observed immunodominance hierarchies among the neoepitope-specific CD8+ TILs, whereby a majority of tetramer-bound CD8+ T cells were restricted to one or two putative neoepitopes. Our results indicate that robust antitumor immune responses are induced in these pediatric ALL tumors despite their low mutation burdens and emphasize the importance of immunodominance in shaping cellular immune responses. These data suggest that pediatric cancers may be amenable to immunotherapies aimed at enhancing immune recognition of tumor-specific neoantigens.
Citation Format: Paul G. Thomas. Hitting the target: T-cell receptors, cross-reactivity, and tumors [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr IA07.
Abstract IA09: Using high-dimensional machine-assisted analysis for biomarkers detection and guidance of immunotherapy to cancer
Authors: Cardenas Luis , Guglietta Silvia , Wrangle John , Rubinstein Mark , Robinson Mark , Krieg Carsten 
Published: June 2021

Abstract
The emergence of immuno-oncology (IO) agents has led to unprecedented successes in the treatment of deadly cancers and sometimes even cures. These agents act by “releasing the brakes” and thereby unleashing the full potential of cells that fight tumors. Sadly, not every patient benefits from these new treatments. We need to know who will benefit and who may not. Here we approached the problem by designing a customized workflow by using high-dimensional single-cell mass cytometry (CyTOF) combined with machine-learning bioinformatics for the in-depth characterization of single immune cells in predicting and monitoring immune responses. The analysis is data driven, can be adapted to high-throughput approaches, and can model arbitrary trial designs such as batch effects and paired designs. We tested our workflow on two studies: a) Predict response to anti-PD-1 immunotherapy in melanoma. In our discovery cohort peripheral blood mononuclear cells (PBMCs) from 20 patients with stage IV melanoma before and after 12 weeks of anti-PD-1 therapy were analyzed. We observed a clear T-cell response on therapy. The most evident difference in responders before therapy was an enhanced frequency of CD14+ CD16+HLA-DRhi classical monocytes. We validated our results using conventional flow cytometry in an independent exploratory cohort of 31 patients before therapy. Finally, we correlated enhanced monocyte frequencies before therapy initiation with clinical response and could show association with lower hazard, extended progression-free and overall survival. b) Monitoring the immune response in anti-PD-1 immunotherapy-refractory non-small cell lung cancer to a novel combination immunotherapy of anti-PD-1 and an IL-15 superagonist. Twenty-one patients with non-small cell lung cancer who mostly were refractory to previous anti-PD-1 treatment received a novel combination therapy of IL-15 superagonist plus anti-PD-1. A response in the CD8+ T cell compartment was observed. Unexpectedly, our high-dimensional approach was able to detect and characterize a strong expansion of innate tumor-reactive effector NK cells starting around day 4 of therapy. Taken together, our unbiased artificial intelligence-driven immune workflow might support patient selection prior to therapy, select the right drug combination, and identify new druggable cell populations.
Citation Format: Luis Cardenas, Silvia Guglietta, John Wrangle, Mark Rubinstein, Mark Robinson, Carsten Krieg. Using high-dimensional machine-assisted analysis for biomarkers detection and guidance of immunotherapy to cancer [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr IA09.
Abstract IA12: Fueling T cells in inflammation and cancer
Authors: Rathmell Jeffrey C. 
Published: June 2021

Abstract
Lymphocyte activation leads to rapid proliferation and differentiation, and we have shown that CD4 T-cell subsets are metabolically distinct. These specific metabolic programs may allow new understanding and approaches to manipulate immunity. Using metabolic network analysis of metabolomics and proteomics, we defined several metabolic nodes differentially utilized by CD4 T-cell subsets, including glutamine metabolism. By genetically deleting the glucose transporter Glut1 or the glutaminolysis enzyme, glutaminase (GLS), we have shown that glycolysis and glutaminolysis are both used by activated T cells. All effector T cells require Glut1 and glycolysis. In contrast, we found that Th17 cells preferentially require GLS while Th1 cells are actively impaired by this enzyme. Thus, inhibition of GLS both reduces Th17 responses and promotes differentiation of Th1 cells and can lead to signs of T-cell exhaustion. We show that GLS deficiency can protect against Th17-mediated inflammatory models and also can augment effector function of Th1 CAR-T cells against B-cell targets. Understanding mechanisms that regulate T-cell metabolism may provide new tools to modulate immunity in the balance of T-cell effector populations to both suppress inflammation and promote effector function.
Citation Format: Jeffrey C. Rathmell. Fueling T cells in inflammation and cancer [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr IA12.
Abstract IA18: The mechanistic basis of cancer immunotherapy
Authors: Mellman Ira 
Published: June 2021

Abstract
The advent of new approaches to the immunotherapy of cancer has caused a dramatic shift not only in the treatment of cancer but also in our understanding of cancer biology. The rapid rate of progress in the clinic, however, has outpaced our understanding of the basic mechanistic features that underlie the therapeutic advances. This is most notable in the case of “checkpoint” inhibitors, such as antibodies to the negative regulatory axis defined by PD-1 and PD-L1. While blocking the interaction of PD-L1 with PD-1 is often assumed to reverse the process of T-cell exhaustion, there is little direct evidence for this interpretation, an incomplete definition of what is meant by “exhaustion,” and a poor understanding of how PD-1 regulates T-cell activity. Starting with observations made in the clinic, we have used biochemical reconstitution together with in vivo analysis in mice to illuminate key features of the PD-L1/PD-1 axis, demonstrating that it functions primarily to regulate T-cell function by controlling signaling by costimulators such as CD28. Conditional knockout experiments also revealed that dendritic cell B7.1 plays a uniquely important role in activating CD28. Distinct from costimulatory molecules, entirely different regulatory mechanisms appear to regulate T-cell receptor signaling. Finally, we have begun to evaluate the origin of T cells that engage antitumor responses, results that further emphasize a need for continuous priming of antitumor T cells to maintain a potent anticancer response.
Citation Format: Ira Mellman. The mechanistic basis of cancer immunotherapy [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr IA18.
Abstract IA19: Lymphatic vessels: Barriers to adaptive immunity in melanoma
Authors: Lund Amanda W. 
Published: June 2021

Abstract
Mechanisms of immune suppression in peripheral tissues counteract protective immunity to prevent immunopathology and are co-opted by tumors for immune escape. Nonhematopoietic cells, as the resident tissue scaffold, play critical roles in regulating the infiltration, retention, function, and exit of tissue- and tumor-infiltrating leukocytes. While lymphatic vessels facilitate T-cell priming through dendritic cell and antigen delivery to lymph nodes, they also facilitate regional metastasis and exert multiple immune-suppressive mechanisms that maintain peripheral tolerance in steady state lymph nodes. We hypothesized that in tumors peripheral lymphatic vessels exhibit context-dependent function that limits local effector CD8+ T-cell accumulation in melanoma. Our work demonstrates that lymphatic vessels both maintain homeostatic mechanisms of peripheral tolerance in poorly inflamed tumors to limit immune surveillance and activate mechanisms of adaptive immune resistance in the context of potent cytotoxic T-cell activity. Consequently, we present tumor-associated lymphatic vessels as a new, anatomic immune checkpoint in melanoma. Our work suggests that understanding context-dependent lymphatic vessel function will continue to reveal novel mechanisms of immune control and metastasis that may be targeted for therapeutic response.
Citation Format: Amanda W. Lund. Lymphatic vessels: Barriers to adaptive immunity in melanoma [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr IA19.