bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract A24: Cancer cells reduce macrophage CXCR3/CXCL10 axis expression through canonical NF-κB signaling
Authors: Korkaya Ahmet K , Bradford Jennifer W 
Published: December 2022

Abstract
Tumor-associated macrophage (TAM) infiltration of tumors via chemokine axis are linked with tumor progression and metastasis. There are forty-seven known chemokines and nineteen known chemokine receptors in humans. Many of these chemokines have a dual functionality in cell survival and growth. Notably, CXCL10 and its receptor CXCR3 are associated with both angiogenesis and angiostasis, among other functions, both pro and anti-tumor. This duality is coupled with the uncertainty of which signaling pathways are responsible for CXCL10 expression and supression during tumor progression. Recent literature states that nuclear factor-kappaB (NF-κB) signaling is required for driving the expression of CXCL10 during traditional macrophage activation; however, little is known about the chemokine’s suppression. To determine the role of NF-κB on CXCR3/CXCL10 modulation in macrophages within the context of cancer, we performed three co-culture experiments to examine the gene expression level changes within bone marrow-derived macrophages (BMDMs) from an NF-κB (p65) deficient mouse model (p65fl/fl/LysMCre) and compared them to control BMDMs (LysMCre). The first co-culture included a murine glioblastoma multiforme (GBM) cell line (GL261), while the other two included different murine triple-negative breast cancer (TNBC) cell lines (AT-3, 4T1). BMDMs, from either LysMCre or p65fl/fl/LysMCre mice, were seeded into 6-well plates, let rest for 24 hours, and then cancer cells were seeded onto the transwell insert. Cells shared the growth medium for 24 hours before collection and subsequent RNA isolation, cDNA synthesis, and qPCR analysis. Here, we identified that BMDM CXCL10 gene expression is drastically reduced when co-cultured with the three different cancer cell lines. Furthermore, we found that conditional deletion of canonical NF-κB signaling (p65) in BMDMs significantly recovered their CXCL10 expression when co-cultured with GL261 glioma cells and AT-3 breast cancer cells. Co-culture with 4T1 breast cancer cells also showed a recovering CXCL10 gene expression in the NF-κB (p65) deficient BMDMs. Additionally, the chemokine’s receptor, CXCR3, gene expression was signficantly reduced in all three co-culture conditions. However, in comparison, there was no significant difference in CXCR3 gene expression in the NF-κB (p65) deficient BMDMs in the same co-cultures. Thus, canonical NF-κB signaling is implicated in maintenance of CXCL10 expression in BMDMs, but not obviously involved in the alteration of their CXCR3 expression.
Citation Format: Ahmet K Korkaya, Jennifer W Bradford. Cancer cells reduce macrophage CXCR3/CXCL10 axis expression through canonical NF-κB signaling [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy; 2022 Oct 21-24; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(12 Suppl):Abstract nr A24.
Abstract A31: CDK4/6 inhibition plus radiotherapy enhances anti-tumor immune responses in non-small cell lung cancer
Authors: Huang Zhengrong , Xie Hongxin , Wang Mengqin , Xie Conghua , Gong Yan 
Published: December 2022

Abstract
Background: Lung cancer is the main cause of cancer-related mortality, of which non-small cell lung cancer (NSCLC) accounts for 85%. Abemaciclib, a selective CDK4/6 inhibitor, enhanced the radiosensitivity of NSCLC in vivo and in vitro and improved the prognosis of advanced NSCLC patients according to a Phase Ⅰ clinical trial. This study aimed to explore the impact of CDK4/6 inhibition combined with irradiation and immunotherapy on NSCLC.Methods: The in vivo xenograft tumor mouse model was used to investigate the synergistic effects of CDK4/6 inhibitor abemaciclib and irradiation together with anti-PD-1 antibodies. The tumor infiltrating lymphocytes (TILs) in tumor microenvironment (TME) were analyzed by flow cytometry. The complete blood count and serum tests were used to determine the safety of this combination therapy. The human phospho-kinase array and dual-luciferase report assay were used to study the regulatory mechanisms of abemaciclib and irradiation combination on PD-L1 transcription in NSCLC cells in vitro.Results: Our results indicated the significantly synergistic effects of abemaciclib, irradiation and anti-PD-1 antibodies on NSCLC growth in vivo, accompanied by increased CD8+ T cell infiltration and decreased regulatory T cells and myeloid-derived suppressor cells (MDSCs), suggesting a strong anti-tumor combination of CDK4/6 inhibition, radiotherapy and immunotherapy. This synergistic effect could be partially impaired by CD8 depletion, and alleviated the anemia and inflammation caused by tumor burden without hepatorenal toxicity. Abemaciclib induced PD-L1 transcription through JNK/c-Jun pathway in vitro, which might elevate PD-L1 expression synergistically with irradiation through TBK1/IRF3 pathway.Conclusions: Our studies suggested that CDK4/6 inhibition plus irradiation enhanced anti-tumor immune responses in NSCLC. Together with immunotherapy, the combination of CDK4/6 inhibitors and radiotherapy could lower the doses of individual treatment, thus alleviating the potential toxicity and side effects of CDK4/6 inhibition and irradiation on normal cells.
Citation Format: Zhengrong Huang, Hongxin Xie, Mengqin Wang, Conghua Xie, Yan Gong. CDK4/6 inhibition plus radiotherapy enhances anti-tumor immune responses in non-small cell lung cancer [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy; 2022 Oct 21-24; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(12 Suppl):Abstract nr A31.
Abstract A37: Evaluation of eDHFR/iTag PET reporter gene immunogenicity and application in GPC3 CAR T cells
Authors: Sellmyer Mark A , Lee Iris K , Kuszpit Kyle , Roy Jyoti , Alfaro Alex , Ory Virginie , Cheng Lily , Sutton Daniel , Bosco Emily , Fazenbaker Christine , Novarra Shabazz , Gilbreth Ryan , Tschernia Nick , Berry Deborah , Chen Xiaoru , Wu Yuling , Wong Ryan 
Published: December 2022

Abstract
Genetically engineered medicines such as chimeric antigen receptor (CAR) T cells have great potential to be the next pillar of medical therapy beyond chemo- and traditional biologic therapies. To develop genetic medicines, new methods to understand their pharmacokinetics (PK) in humans are crucial. It is not feasible to perform traditional PK analysis for “living drugs”, because the genes themselves (in the form of DNA or RNA), are not typically responsible for the therapeutic effect. Rather, the protein products of the genes or the cells harboring the engineered genes are the actuators, and thus cannot be measured using standard HPLC or ligand binding immunoassays for PK analysis. We used a positron emission tomography (PET) reporter gene or “imaging tag” based on the intracellular bacterial enzyme dihydrofolate reductase (eDHFR) that can be paired with radiolabeled versions of trimethoprim (TMP). In this work, we evaluate the potential for immunogenicity using primary human cells and assays geared to assess low affinity and rare T cell clones that may react to eDHFR. We used overlapping pools of 15-mer eDHFR peptides and found that across 9 patients, there was little reactivity compared to EBV and CMV peptide controls. Further, the relative strength of reactivity to the eDHFR peptides was less than that of the viral peptides. Next, we showed that eDHFR iTag harboring CAR T cells were functionally comparable to unlabeled CAR T cells in vitro, and demonstrated strong, selective [18F]-TMP uptake in the eDHFR-expressing CAR T cells. Finally, using a glypican 3 (GPC3) CAR T rodent model, we performed a feasibility study to non-invasively track proliferation in antigen-harboring xenograft tumors over time with ex vivo correlation to anti-CD3 immunohistochemistry. These data demonstrate the potential for non-invasive monitoring of CAR T cells using PET imaging and translational applicability of DHFR/TMP radiotracers.
Citation Format: Mark A Sellmyer, Iris K Lee, Kyle Kuszpit, Jyoti Roy, Alex Alfaro, Virginie Ory, Lily Cheng, Daniel Sutton, Emily Bosco, Christine Fazenbaker, Shabazz Novarra, Ryan Gilbreth, Nick Tschernia, Deborah Berry, Xiaoru Chen, Yuling Wu, Ryan Wong. Evaluation of eDHFR/iTag PET reporter gene immunogenicity and application in GPC3 CAR T cells [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy; 2022 Oct 21-24; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(12 Suppl):Abstract nr A37.
Abstract A49: Proof of principle that loss of mismatch repair protein reduces tumor burden in mouse model of gastric cancer
Authors: Huber Anne , Dijkstra Christine , Whitehall Vicki , Ernst Matthias , Eissmann Moritz 
Published: December 2022

Abstract
Gastric cancer (GC) remains the third leading cause of cancer-related death worldwide. Unfortunately, only a subset of GC patients, characterized by tumors with high microsatellite instability (MSI), responds to immune checkpoint (ICI) therapy. In the context of GC, the mechanisms of how MSI improves ICI therapy responses remain poorly understood. Here we are undertaking a proof of principle study to demonstrate in novel GC mouse models that loss of mismatch repair protein MLH1 confers MSI phenotype and impairs tumor growth via altered anti-tumor immune responses. To study the functional and mechanistic effects of loss of MLH1 protein, we established MLH1-deficient (Kras-, Pi3kca-, Tp53-mutant) murine tumor organoids via CRISPR/Cas9 technology. These organoids were subcutaneously allografted into immunocompetent C57BL/6 mice to study tumor progression, immune surveillance, and responses to immunotherapy in vivo. MSI testing confirmed that MLH1-proficient parental organoids are MSI low, whereas MLH1-deficient organoids are MSI high. Low passage MLH1-deficient organoid tumors grew similarly to MLH1-proficient tumors. However, after culturing of organoids in vitro for a prolonged time prior to injection to allow accumulation of mutations, subcutaneous allograft MLH1-deficient tumors were considerably smaller compared to MLH1-proficient tumors. MLH1-deficient tumors showed a significantly higher number of CD8+ T cells. Additionally, MLH1-deficient tumors were similar in size to MLH1-proficient tumors when allografted into Rag1-/- mice. Interestingly, CD8- and CD4-depleting experiments showed an involvement of CD4+ T cells in the impairment of MLH1-deficient tumor growth. Treatment with anti-PD-1, but not anti-CTLA-4, reduced tumor mass further. We are currently investigating the underlying mechanism for the impaired growth of MLH1-deficient organoid allograft tumors via tumor mutational burden analysis and neoantigen testing. Taken together, we provide evidence that loss of MLH1 leads to high MSI in gastric tumors, reduces tumor growth after prolonged in vitro culture and increases response to anti-PD-1 therapy. Our findings encourage further studies to investigate the mechanisms of impaired tumor growth after MLH1 loss in GC and may provide insights leading to improve ICI therapy responses for GC patients.
Citation Format: Anne Huber, Christine Dijkstra, Vicki Whitehall, Matthias Ernst, Moritz Eissmann. Proof of principle that loss of mismatch repair protein reduces tumor burden in mouse model of gastric cancer [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy; 2022 Oct 21-24; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(12 Suppl):Abstract nr A49.
Abstract A58: Increased systemic activated CD4+ T cells and non-classical monocytes are associated with improved immunotherapy response and progression-free survival in non-small cell lung cancer patients
Authors: Figueiredo Amanda B , Ferreira Stephanie M Iasbeke , Evangelista Guilherme FB , Santos Iasmim P , Kuil Larissa M , Cavalcanti Clara M , Morais Kátia LP , Galdino Nayane AL , Lopes Ananda D , Freitas Helano C , Gross Jefferson L , Pinto Clóvis A Lopes , Chojniak Rubens , de Lima Vladmir C Cordeiro , Gollob Kenneth J 
Published: December 2022

Abstract
The development of immune checkpoint inhibitors initiated a new era of anti-cancer therapies with very promising results. Although effective for many patients, others are primarily refractory to this treatment. Identifying systemic immune cell subpopulations associated with treatment success or failure can reveal mechanisms behind therapeutic failure and offer potential predictive markers of clinical use. Here, for the first time, we performed a prospective non-interventional study and evaluated the pretreatment systemic immunological status of 33 patients with stage IV non-small cell lung cancer treated with anti-PD-1 combined with chemotherapy by flow cytometry. Treatment outcome was evaluated 9 weeks after treatment to define patients as Responders and Non-responders. Patients were reevaluated 6 months after therapy. Responders presented a higher frequency of activated CD4+ CD69+ T lymphocytes and non-classical monocytes before treatment began, which acted as predictive biomarkers for response measured at nine weeks. Conversely, CTLA-4+CD4+ T lymphocytes were more frequent in Non-responders and may contribute to an immunosuppressive milieu impairing the control of the tumor. Patients with durable responses (³ 6 months) also had higher frequency of activated CD4+ T lymphocytes before treatment started, compared with non-responders. In contrast, non-classical monocytes did not predict a durable response. However, patients with a higher frequency of both immune populations had longer median progression free survival. Our results suggest that higher frequency of peripheral activated CD4+CD69+, CD4+CTLA-4+ T cells may be useful biomarkers of therapeutic success or failure in NSCLC and point to mechanisms underlying therapeutic outcome. Studies are underway to further characterize the functional potential and activation status these systemic immune subpopulations.
Citation Format: Amanda B Figueiredo, Stephanie M Iasbeke Ferreira, Guilherme FB Evangelista, Iasmim P Santos, Larissa M Kuil, Clara M Cavalcanti, Kátia LP Morais, Nayane AL Galdino, Ananda D Lopes, Helano C Freitas, Jefferson L Gross, Clóvis A Lopes Pinto, Rubens Chojniak, Vladmir C Cordeiro de Lima, Kenneth J Gollob. Increased systemic activated CD4+ T cells and non-classical monocytes are associated with improved immunotherapy response and progression-free survival in non-small cell lung cancer patients [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy; 2022 Oct 21-24; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(12 Suppl):Abstract nr A58.
Abstract B12: T-cell derived extracellular vesicles prime macrophages for improved STING based cancer immunotherapy
Authors: Hansen Aida S. , Jensen Lea S. , Ryttersgaard Kristoffer G. , Krapp Christian , Just Jesper , Joensson Kasper L. , Etzerodt Anders , Deleuran Bent W. , Jakobsen Martin R. 
Published: December 2022

Abstract
Despite recent years advancement reported for cancer immunotherapies within multiple cancer types, we still see a large group of patients failing to respond to treatment. Activation of the innate immune pathway controlled by STimulator of INterferon Genes (STING) has been considered as an attractive target for boosting cancer immunotherapy. Extracellular vesicles (EVs) are key players in shaping immune responses and are proposed to provide a danger signal for antigen-presenting cells. Here, we hypothesize that CD4+ T-cell derived EVs (T-EVs) enhance innate immune cells function, such as macrophages, by priming the STING signaling pathway. We purified T-EVs from aCD3/aCD28 stimulated CD4+ T cells by differential ultracentrifugation, and primed macrophages with these prior to stimulation with STING agonists. Subsequent activation was measured by induction of type I Interferons and phosphorylation of signaling molecules within the STING pathway. We further explored the in vivo efficacy of combining T-EVs priming and STING agonists in tumor bearing mice models. We found that T-EVs sensitizes macrophages to respond more potently to STING activation. Importantly, this was independent on both surface-associated and intravesicular DNA. The priming of STING signaling was largely induced by IFNγ and TNFα carried by the T-EVs. We further showed that the T-EVs enhances the efficacy of STING agonist stimulation, by controlling tumor growth in syngeneic MC38 tumor models. Our work support that T-EVs can disrupt the immune suppressive tumor microenvironment by reprogramming macrophages to a pro-inflammatory phenotype, and priming them for a robust immune response towards STING activation.
Citation Format: Aida S. Hansen, Lea S. Jensen, Kristoffer G. Ryttersgaard, Christian Krapp, Jesper Just, Kasper L. Joensson, Anders Etzerodt, Bent W. Deleuran, Martin R. Jakobsen. T-cell derived extracellular vesicles prime macrophages for improved STING based cancer immunotherapy [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy; 2022 Oct 21-24; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(12 Suppl):Abstract nr B12.
Abstract B17: Data-driven discovery of targets for immune-metabolic antitumor drugs identifies Estrogen Related Receptor Alpha
Authors: Sahu Avinash 
Published: December 2022

Abstract
Drugs that kill tumors through multiple mechanisms have the potential for broad clinical benefits. Here, we first developed an in silico multi-omics approach (BipotentR) to find cancer-cell-specific regulators that simultaneously modulate tumor immunity and another oncogenic pathway, and use it to identify 38 candidate immune-metabolic regulators. We show the tumor activity of these regulators stratify melanoma patients by their response to anti-PD1 using machine learning and deep neural approaches, which we benchmarked against current biomarkers. The topmost identified regulator, ESRRA (Estrogen Related Receptor Alpha), is activated in immunotherapy-resistant tumors of many types. Its inhibition killed tumors by suppressing energy metabolism and activating two immune mechanisms: (i) cytokine induction, causing proinflammatory macrophage polarization (ii) antigen-presentation stimulation, recruiting CD8+ T cells into tumors. We also demonstrate a wide utility of BipotentR by applying it to angiogenesis and growth-suppressor pathways. BipotentR, available at http://bipotentr.dfci.harvard.edu/, provides a resource for evaluating patient response and discovering drug targets that act simultaneously through multiple mechanisms.
Citation Format: Avinash Sahu. Data-driven discovery of targets for immune-metabolic antitumor drugs identifies Estrogen Related Receptor Alpha [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy; 2022 Oct 21-24; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(12 Suppl):Abstract nr B17.
Abstract B28: Targeting TBK1 to overcome resistance to cancer immunotherapy
Authors: Sun Yi , Revach Or-yam , Anderson Seth , Manguso Robert T. , Jenkins Russell W 
Published: December 2022

Abstract
Despite the success of PD-1 blockade in melanoma and other cancers, effective treatment strategies to overcome resistance to cancer immunotherapy are lacking. We identified the innate immune kinase TANK-binding kinase 1 (TBK1) as a candidate immune evasion gene in a pooled genetic screen. Using a suite of genetic and pharmacologic tools across multiple experimental model systems, we confirm a role for TBK1 as an immune evasion gene. Targeting TBK1 enhances response to PD-1 blockade by lowering the cytotoxicity threshold to effector cytokines (TNF𝛂/IFN𝛄). TBK1 inhibition in combination with PD-1 blockade also demonstrated efficacy using patient-derived tumor models, with concordant findings in matched patient-derived organotypic tumor spheroids (PDOTS) and matched patient-derived organoids (PDOs). Tumor cells lacking TBK1 are primed to undergo RIPK- and caspase-dependent cell death in response to TNF𝛂/IFN𝛄 in a JAK/STAT-dependent manner. Taken together, our results demonstrate that targeting TBK1 is a novel and effective strategy to overcome resistance to cancer immunotherapy.
Citation Format: Yi Sun, Or-yam Revach, Seth Anderson, Robert T. Manguso, Russell W Jenkins. Targeting TBK1 to overcome resistance to cancer immunotherapy [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy; 2022 Oct 21-24; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(12 Suppl):Abstract nr B28.
Abstract B40: Co-targeting autophagy, macrophages and vasculature in glioma tumors triggers tumor immunity
Authors: Chryplewicz Agnieszka , Scotton Julie , Tichet Mélanie , Hanahan Douglas 
Published: December 2022

Abstract
Glioblastoma (GBM) is the most common primary tumor arising in the central nervous system (CNS). The currently approved standard of care has transient clinical benefit as GBM tends to be exceedingly aggressive. Despite pronounced efforts to identify novel therapies, curative options for GBM do not exist, and the survival rate of diagnosed patients is very low. Therefore, there is an urgent need for new treatment strategies. One approach would be to co-target and thereby disrupt distinct hallmarks of cancer, aiming to elicit sustained therapeutic responses. We have assessed this concept by combining a tricyclic antidepressant -imipramine - with drugs targeting VEGF-A ligand or VEGF-Receptor in mice bearing de novo GBM. All monotherapies were ineffective. In notable contradistinction, we found that combinatorial regimens significantly increased survival benefit and regressed established tumors. Investigation of the basis for the therapeutic efficacy revealed that combining the VEGF pathway inhibitor with imipramine accentuated autophagy while modifying the angiogenic tumor vasculature to be more normal-like with induction of high endothelial venules. In addition, imipramine downregulated an M2-like phenotype of tumor-associated macrophages via histamine receptor and reprogramed them to express chemokines attracting otherwise rare CD8 T cells, which demonstrably contributed to the observed efficacy. As such, these hallmark co-targeting combinations reprogram the GBM microenvironment from immunosuppressive to pro-inflammatory, thereby rendering it immunogenic and sensitizing the tumors to immune checkpoint blockade, as evidenced by enhanced responses when an anti-PD-L1 therapy was included in the mix. The results to be presented will elaborate on a provocative new therapeutic approach for glioblastoma that has the prospect of motivating clinical evaluation in this daunting form of human cancer.
Citation Format: Agnieszka Chryplewicz, Julie Scotton, Mélanie Tichet, Douglas Hanahan. Co-targeting autophagy, macrophages and vasculature in glioma tumors triggers tumor immunity [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy; 2022 Oct 21-24; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(12 Suppl):Abstract nr B40.