bluuebunny/crossref_metadata_2025 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

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

Introduction: Immunotherapy is promising for patients with treatment-refractory head and neck squamous cell carcinoma (HNSCC). However, >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

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

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

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

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

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

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

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.
Abstract IA24: Immune-modulating approaches to cancer therapy: Focusing on combinations
Authors: Wolchok Jedd
Published: June 2021

Given the activity noted with both CTLA-4 or PD-1 blockade, clinical trials are now investigating combination checkpoint blockade. The most mature data with a combination of ipilimumab + nivolumab in melanoma showed a response rate of 60% in the context of increased yet manageable toxicity. Such responses are generally durable, even when treatment was stopped early for toxicity. Unlike in studies of PD-1 blockade monotherapy, there was no significant difference in clinical activity based on tumor expression of PD-L1. This approach has gained regulatory approval for metastatic melanoma and is in late-stage clinical trials for other malignancies. Attention is being paid to the reasons underlying the efficacy of checkpoint blockade in certain malignancies. One hypothesis has been that cancers having a high mutational load may be more amenable to immune modulation by virtue of the larger number of potential neoepitopes present, fostering baseline immune recognition that can then be potentiated by checkpoint blockade. We have found that melanoma patients having long-term clinical activity with ipilimumab have a significantly greater median number of nonsynonymous passenger mutations, compared with patients who do not respond or those who have only short-term regression. Strategies to enhance baseline immune reactivity are therefore necessary to investigate as a means to improve the impact of checkpoint blockade on a broad spectrum of cancers. The presence of suppressive myeloid cells in the tumor microenvironment also is emerging as a mechanism of resistance to the antitumor activity for checkpoint blockade. Strategies to overcome this include inhibition of CSF-1R signaling, IDO activity, and selective suppression of PI3K-γ.

Citation Format: Jedd Wolchok. Immune-modulating approaches to cancer therapy: Focusing on combinations [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 IA24.
Abstract IA25: Control of tumor immune suppression by macrophage PI3Kγ regulated pathways
Authors: Varner Judith A.
Published: June 2021

Macrophages play key roles in promoting immune suppression, tumor growth, and resistance to therapy. Our recent studies show that tissue-resident macrophages (TRM) as well as bone marrow-derived macrophages (BMDM) play critical but unique roles in promoting tumor growth. While BMDM are recruited to tumors in a PI3Kγ, Integrin α4, and CCR2-dependent manner, TRMs accumulate in tumors independently of integrin α4, PI3Kγ, or the trafficking receptor CCR2. Gene expression and functional studies indicate that tumor-derived TRM are highly proliferative, immune suppressive, and distinct from BMDM. We show that TRM develop from macrophage progenitors that are abundant in tumors but not in normal tissues; purified progenitors differentiate into macrophages that potently stimulate tumor growth when adoptively transferred into mice. Notably, we found that the lipid kinase PI3Kγ promotes immune-suppressive polarization of TRM and BMDM. Inhibition of PI3Kγ repolarizes macrophages, induces dendritic cell maturation, and stimulates CD8+ T-cell recruitment to tumors and activation. Inhibition of PI3Kγ also synergizes with checkpoint inhibitors and chemotherapeutics to promote tumor regression and immunologic memory, leading to sustained tumor suppression. Our studies identify PI3Kγ as a valuable target for immune therapy of solid tumors.

Citation Format: Judith A. Varner. Control of tumor immune suppression by macrophage PI3Kγ regulated pathways [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 IA25.
Abstract PR04: Single-cell analysis illuminates dysfunctional CD8+ T cells as a proliferative, dynamically regulated compartment within human melanoma
Authors: Yofe Ido , Li Hanjie , van der Leun Anne , Lubling Yaniv , Solodkin Dikla Gelbard , van Akkooi Alexander , Horlings Hugo M. , David Eyal , Baran Yael , Berkovitz Akihad , Lifshitz Aviezer , Schumacher Ton N. , Tanay Amos , Amit Ido
Published: June 2021

The immune cell composition within tumors plays a major role in response to immunotherapy. Therefore, comprehensive characterization of the diverse functional states exhibited by immune cell infiltrates, and their association with treatment outcome, is critical for the development of more effective immunotherapies. We combined single-cell RNA- and TCR-sequencing with an autologous tumor reactivity assay, tracing immune cell heterogeneity and dynamics in melanoma patients. Analysis of 70,000 cells revealed the presence of two effector T-cell subsets, of which only one transitions into a dysfunctional T-cell population, as based both on TCR sharing as well as a gradient of expression of dysfunction-associated genes present in virtually all samples. Notably, we denote dysfunctional T cells— and in particular early dysfunctional cells—as the major proliferating immune cell compartment. Furthermore, TCR-seq of autologous tumor reactive T cells illustrated that presence of reactive potential is associated with the magnitude of dysfunctional signature and is in concordance with response to immunotherapy. Data from 25 melanoma patients confirm the universality of these observations and distinguish shared and divergent regulatory modules between dysfunctional CD8 and regulatory T cells. Our analysis also puts forward novel candidate genes, which are highly correlated with known checkpoint targets within specific cellular subsets and may prove to be effective targets for checkpoint blockade. In conclusion, our findings proclaim that CD8 T cells previously associated with a dysfunctional or exhausted state are in fact a highly proliferating and dynamically interacting cell population within the human tumor microenvironment.

This abstract is also being presented as Poster B58.

Citation Format: Ido Yofe, Hanjie Li, Anne van der Leun, Yaniv Lubling, Dikla Gelbard Solodkin, Alexander van Akkooi, Hugo M. Horlings, Eyal David, Yael Baran, Akihad Berkovitz, Aviezer Lifshitz, Ton N. Schumacher, Amos Tanay, Ido Amit. Single-cell analysis illuminates dysfunctional CD8+ T cells as a proliferative, dynamically regulated compartment within human 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 PR04.
Abstract PR05: Blockade of AHR activation by IDO/TDO-derived kynurenine restricts cancer immune suppression
Authors: Campesato Luis F. , Budhu Sadna , Gigoux Mathieu , Tchaicha Jeremy , Pourpe Stephane , Liu Cailian , Zamarin Dmitriy , Manfredi Mark G. , McGovern Karen , Wolchok Jedd D. , Merghoub Taha
Published: June 2021

Immune checkpoint blockade (ICB) has been shown to convey significant clinical activity across a spectrum of malignancies, yet there is now recognition that multiple mechanisms of resistance can impair response. The catabolism of tryptophan into metabolites known as kynurenines (Kyn) by enzymes such as indoleamine 2,3-dioxygenase (IDO) or tryptophan 2,3-dioxygenase (TDO) plays a major suppressive role. Recently it was shown that Kyn acts as an endogenous agonist of the Aryl hydrocarbon receptor (AhR). In order to gain a better understanding of this pathway, we sought to characterize the mechanisms of immunosuppression associated with AhR and evaluate its potential as therapeutic target. Gene-expression analysis of IDO-overexpressing melanomas (B16-IDO vs. B16-WT) demonstrated reduced expression levels of Type 1 inflammatory genes, including IFNy, TNF, GzmB, and CD40. In addition, B16-IDO presents higher infiltration of tumor-associated macrophages TAMs, which upregulate the AHR as well as classic AhR-regulated genes (Cyp1a1 and Cyp1b1) and are differentially skewed towards an immunosuppressive M2 phenotype. Tumor-antigen specific CD8+T cells show reduced expression of activation markers (GzmB and CD44) and proliferation rate when primed by Kyn-treated antigen-presenting cells. In addition, TAMs from B16-IDO tumors suppressed activation of CD8+T cells in vitro and their depletion delayed tumor growth. When B16-IDO cells are implanted in mice depleted of Foxp3 expressing cells, TAMs do not accumulate. Treatment of B16-IDO tumors with an AhR-specific antagonist (CH-223191) upregulates MHC II in APCs, activation markers in CD8s, and reduced frequency of T-regs in B16-IDO tumors. AhR inhibition slows tumor growth and prolongs survival of tumors with active IDO/TDO/Kyn pathway (B16-IDO and B16-TDO), and this is enhanced when PD-1 blockade is used in combination. In summary, our findings demonstrate that targeting the Kyn pathway through AhR inhibition could overcome key suppressive mechanisms and sensitize tumors to ICB.

This abstract is also being presented as Poster A57.

Citation Format: Luis F. Campesato, Sadna Budhu, Mathieu Gigoux, Jeremy Tchaicha, Stephane Pourpe, Cailian Liu, Dmitriy Zamarin, Mark G. Manfredi, Karen McGovern, Jedd D. Wolchok, Taha Merghoub. Blockade of AHR activation by IDO/TDO-derived kynurenine restricts cancer immune suppression [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 PR05.
Abstract PR22: Collagen scavenging alters macrophage phenotype in pancreatic cancer
Authors: LaRue Madeleine M. , Cullis Jane , Bar-Sagi Dafna
Published: June 2021

A hallmark of pancreatic tumors is their highly desmoplastic stroma composed of fibroblasts, immune cells and a dense network of collagen fibers. Pancreatic tumor-associated macrophages are considered M2 macrophages and are thought to support tumor progression through the engagement of multiple mechanisms that promote immune tolerance. Another well-documented role of M2 macrophages is the remodeling of the tumor stroma via collagen production and degradation, with the latter being mediated by mannose receptor (Mrc1)-mediated endocytosis of collagen. Given our previous work showing that protein scavenging by tumor cells can be utilized as a nutrient delivery mechanism, we set out to investigate the functional significance of collagen scavenging by macrophages. We have found that, in M2 macrophages, Mrc1-mediated collagen internalization and subsequent lysosomal degradation leads to a metabolic reprogramming that is characterized by the accumulation of collagen-derived intracellular free amino acids and an increase in arginine biosynthesis. The increase in arginine levels results in a significant upregulation of inducible nitric oxide synthase (iNOS) and arginase 1 (ARG1), giving rise to an M1/M2 hybrid macrophage. Moreover, iNOS+ARG1+ double-positive macrophages are present in the microenvironment of pancreatic tumors. The functional contribution of this cell population to pancreatic tumorigenesis is being investigated. Our findings suggest a new modality for the functional modulation of tumor-associated macrophages through the coupling of extracellular matrix remodeling to protein scavenging.

This abstract is also being presented as Poster A58.

Citation Format: Madeleine M. LaRue, Jane Cullis, Dafna Bar-Sagi. Collagen scavenging alters macrophage phenotype in pancreatic 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 PR22.
Prevention of Growth Failure in Turner Syndrome: Long-Term Results of Early Growth Hormone Treatment in the “Toddler Turner” Cohort
Authors: Quigley Charmian A. , Fechner Patricia Y. , Geffner Mitchell E. , Eugster Erica A. , Ross Judith L. , Habiby Reema L. , Ugrasbul Figen , Rubin Karen , Travers Sharon , Antalis Caryl J. , Patel Hiren N. , Davenport Marsha L.
Published: June 2021

Introduction: In the randomized “Toddler Turner” study, girls who received growth hormone (GH) starting at ages 9 months to 4 years (early-treated [ET] group) had marked catch-up growth and were 1.6 ± 0.6 SD taller than untreated (early-untreated [EUT]) control girls after 2 years. However, whether the early catch-up growth would result in greater near-adult height (NAH) was unknown. Therefore, this extension study examined the long-term effects of toddler-age GH treatment on height, pubertal development, and safety parameters. Methods: Toddler Turner study participants were invited to enroll in a 10-year observational extension study for annual assessments of growth, pubertal status, and safety during long-term GH treatment to NAH for both ET and EUT groups. Results: The ET group was taller than the EUT group at all time points from preschool to maturity and was significantly taller at the onset of puberty (p = 0.016), however, the difference was not significant at NAH. For the full cohort (ET + EUT combined, n = 50) mean (± SD) NAH was 151.2 ± 7.1 cm at age 15.0 ± 1.3 years. NAH standard deviation score (SDS) was within the normal range (>−2.0) for 76% of ET and 60% of EUT subjects (68% overall) and correlated strongly with height SDS at GH start (r = 0.78; p < 0.01), which in turn had a modest inverse correlation with age at GH start (i.e., height SDS declined with increasing age in untreated girls [r = −0.30; p = 0.016]). No new safety concerns arose. Conclusion: Although the ET group was taller throughout, height SDS at NAH was not significantly different between groups due to catch-down growth of ET girls during lapses in GH treatment after the Toddler study and similar long-term GH exposure overall. Early initiation of GH by age 6 years, followed by uninterrupted treatment during childhood, can prevent ongoing growth failure and enable attainment of height within the normal range during childhood, adolescence, and adulthood.
Effects of Finerenone Combined with Empagliflozin in a Model of Hypertension-Induced End-Organ Damage
Authors: Kolkhof Peter , Hartmann Elke , Freyberger Alexius , Pavkovic Mira , Mathar Ilka , Sandner Peter , Droebner Karoline , Joseph Amer , Hüser Jörg , Eitner Frank
Published: June 2021

Introduction: The nonsteroidal mineralocorticoid receptor (MR) antagonist finerenone and sodium-glucose cotransporter-2 (SGLT2) inhibitors have demonstrated clinical benefits in CKD patients with type 2 diabetes. Clinical data analyzing the potential value of a combination therapy are currently limited. We therefore investigated cardiorenal protection of respective mono- and combination therapy in a preclinical model of hypertension-induced end-organ damage. Methods: Cardiovascular (CV) morbidity and mortality were studied in hypertensive, N(ω)-nitro-L-arginine methyl ester-treated, renin-transgenic (mRen2)27 rats. Rats (10- to 11-week-old females, n = 13–17/group) were treated once daily orally for up to 7 weeks with placebo, finerenone (1 and 3 mg/kg), empagliflozin (3 and 10 mg/kg), or a combination of the respective low doses. Key outcome parameters included mortality, proteinuria, plasma creatinine and uric acid, blood pressure, and cardiac and renal histology. Results: Placebo-treated rats demonstrated a 50% survival rate over the course of 7 weeks. Drug treatment resulted in variable degrees of survival benefit, most prominently in the low-dose combination group with a survival benefit of 93%. Monotherapies of finerenone or empagliflozin dose-dependently reduced proteinuria, while low-dose combination revealed an early, sustained, and over-additive reduction in proteinuria. Empagliflozin induced a strong and dose-dependent increase in urinary glucose excretion which was not influenced by finerenone coadministration in the combination arm. Low-dose combination but not respective low-dose monotherapies significantly reduced plasma creatinine and plasma uric acid after 6 weeks. Treatment with finerenone and the low-dose combination significantly decreased systolic blood pressure after 5 weeks. There was a dose-dependent protection from cardiac and kidney fibrosis and vasculopathy with both agents, while low-dose combination therapy was more efficient than the respective monotherapy dosages on most cardiorenal histology parameters. Discussion/Conclusions: Nonsteroidal MR antagonism by finerenone and SGLT2 inhibition by empagliflozin confer CV protection in preclinical hypertension-induced cardiorenal disease. Combination of these 2 independent modes of action at low dosages revealed efficacious reduction in important functional parameters such as proteinuria and blood pressure, plasma markers including creatinine and uric acid, cardiac and renal lesions as determined by histopathology, and mortality indicating a strong potential for combined clinical use in cardiorenal patient populations.
Estimation of avoidable losses caused by Meloidogyne incognita infecting cucumber in poly-house
Authors: Bhati Shakti Singh , Baheti B. L.
Published: June 2021

Cucumber (Cucumis sativus L.) is a high nutritious and mineral-rich vegetable, which occupies a prominent place as a salad and vegeta-ble. It is being used in many ways in the daily diet of humans and widely cultivated worldwide. The decrease of agricultural land, ad-verse environmental conditions and continuous increase of popula-tion, the demand of nutritious food is a matter of great concern to the world. Protected cultivation is a very effective tool to solve this problem because in this cultivation the productivity of crops is very high as compared to open field conditions. High value crops suc-cessfully grown in protected cultivation, specially vegetables (cu-cumber, tomato, Capsicum etc.) which are highly susceptible to the numerous pests and pathogens, including phyto-parasitic nema-todes (specially root-knot nematode, Meloidogyne spp.). With this view, present trial was taken to estimate the avoidable losses caused by Meloidogyne incognita infecting cucumber in poly-house situated on farmer’s field with the application of phorate at 2 kg a.i. ha-1 over check. Results exhibited that application of chemical treatment significantly reduced number of galls per 5 g roots, egg masses per 5 g roots, eggs & juveniles per egg mass and final nema-tode population 79.03, 81.10, 30.91 and 56.54%, respectively. Avoidable yield losses were recorded to the tune of 66.84% on cu-cumber by M. incognita in poly-house.
Assessing impact of saline intrusion on rice cultivating area in Ke Sach district, Soc Trang province, Vietnam
Authors: Giao Nguyen Thanh
Published: June 2021

This study was carried out to evaluate the effects of saline intrusion on agricultural activities, thereby proposing adaptation and mitigation measures for local people. Salinity was measured at 15 locations on the main and tributary rivers in Ke Sach district, Soc Trang province, Vietnam from January to September 2020. The method of direct interview with farmers was used to evaluate the knowledge and effects of saline intrusion on agricultural production. Salinity measurement results showed that salinity intrusion oc-curred and lasted from February to the end of May during the dry season (the Winter-Spring rice crop). During the rainy season (June-September), due to the large amount of water, salinity was only be-tween 0.01 and 0.1‰. However, the research has found significant effects in the Summer-Autumn rice crop due to the accumulation of salinity in the soil from the Winter-Spring rice crop, which reduces the rice yield by 10-30%. The interview results revealed that more than 80% of the farmers were aware of the effects of saline water intrusion. Local authorities also regularly organize seminars to disseminate basic knowledge to the community to minimize the risks and impacts of saltwater intrusion. The proposed temporary solutions include rainwater storage, maintenance of saline prevention sluices, construction of more water regulation canals, financial support, and strengthening of salinity monitoring stations. In order to adapt to the long-term salinity conditions, local authorities need to focus on researching changing cropping patterns, crop structure, using new rice varieties that are able to adapt to drought and salty conditions. Besides, there will be technical assistance, capital for people to adapt well to the saline intrusion situation.
Desert locust (Schistocerca gregaria Forskal) and its management: A review
Authors: Katel Shambhu , Mandal Honey Raj , Neupane Pooja , Timsina Sandipa , Pokhrel Pratima , Katuwal Amrit , Subedi Sudeep , Shrestha Jiban , Shah Kabita Kumari
Published: June 2021

The desert locust [Schistocerca gregaria (Forskal)] (Orthoptera: Acrididae) is one of the world’s most dangerous and destructive migratory pests. It is a highly voracious and polyphagous insect. Its swarms can travel hundreds of kilometers per day and infest areas covering millions of square kilometers, resulting in substantial crop losses. We synthesize published research data and reports on the life cycle, behavior, phases, and damage of the desert locust, as well as its possible management practices, in this study. There are diffi-culties involved in estimating locust populations because locust swarms are highly mobile. Cultural control, baiting, dusting with insecticide, and spraying liquid insecticides (chemical or biological) using ultra-low volume (ULV) application are all options for control-ling locust swarms. Improved knowledge on pest biology, as well as more efficient monitoring and control strategies, are essential com-ponents of an effective management strategy.
Racial and Ethnic Disparities in Peripheral Artery Disease
Authors: Hackler Eddie L. , Hamburg Naomi M. , White Solaru Khendi T.
Published: June 2021

Peripheral artery disease is an obstructive, atherosclerotic disease of the lower extremities causing significant morbidity and mortality. Black Americans are disproportionately affected by this disease while they are also less likely to be diagnosed and promptly treated. The consequences of this disparity can be grim as Black Americans bear the burden of lower extremity amputation resulting from severe peripheral artery disease. The risk factors of peripheral artery disease and how they differentially affect certain groups are discussed in addition to a review of pharmacological and nonpharmacological treatment modalities. The purpose of this review is to highlight health care inequities and provide a review and resource of available recommendations for clinical management of all patients with peripheral artery disease.
Vascular Impact of Cancer Therapies: The Case of BTK (Bruton Tyrosine Kinase) Inhibitors
Authors: Fleming Matthew R. , Xiao Ling , Jackson Klarissa D. , Beckman Joshua A. , Barac Ana , Moslehi Javid J.
Published: June 2021

Novel targeted cancer therapies have revolutionized oncology therapies, but these treatments can have cardiovascular complications, which include heterogeneous cardiac, metabolic, and vascular sequelae. Vascular side effects have emerged as important considerations in both cancer patients undergoing active treatment and cancer survivors. Here, we provide an overview of vascular effects of cancer therapies, focusing on small-molecule kinase inhibitors and specifically inhibitors of BTK (Bruton tyrosine kinase), which have revolutionized treatment and prognosis for B-cell malignancies. Cardiovascular side effects of BTK inhibitors include atrial fibrillation, increased risk of bleeding, and hypertension, with the former 2 especially providing a treatment challenge for the clinician. Cardiovascular complications of small-molecule kinase inhibitors can occur through either on-target (targeting intended target kinase) or off-target kinase inhibition. We will review these concepts and focus on the case of BTK inhibitors, highlight the emerging data suggesting an off-target effect that may provide insights into development of arrhythmias, specifically atrial fibrillation. We believe that cardiac and vascular sequelae of novel targeted cancer therapies can provide insights into human cardiovascular biology.