bluuebunny/crossref_metadata_2025_split huggingface dataset:
Integrated profiling of T cells and tumor cells demonstrates metabolic adaptation essential for melanoma cell therapy
Authors: Martinez-Paniagua Melisa A , Haymaker Cara , Adolacion Jay R T , An Xingyue , Bandey Irfan N , Creasy Caitlin , Geiger Tamar , Harel Michal , Robinson Jonathan , Nielsen Jens B , Amritkar Amit , Woodman Scott , Roysam Badrinath , Hwu Patrick , Bernatchez Chantale , Varadarajan Navin 
Published: January 2023

Abstract
T cell-based therapies have shown promising results, but adoptive T cell therapy (ACT) for solid tumors like melanoma treatment has met limited success. The goal of this study was to identify attributes of T cell fitness important for optimal anti-tumor efficacy. This was accomplished by integrated profiling of expanded tumor infiltrating lymphocytes, TILs (responders (CR) and progressive diseases (PD)) co-cultured overnight with their autologous primary tumor cells with the aid of a suite of single-cell, transcriptional, proteomic and functional assays to construct a genome-scale metabolic model of the metabolism of TILs and tumor cells in direct competition with each other. Sorted live TILs after co-culture were used to performed RNA-seq, and proteome-wide profiling by mass spectrometry. Gene enrichment of the CR TIL suggested an increased expression in genes involving processes such as glycolysis, hypoxia, adipogenesis, and mTORC1 signaling. PD tumors showed an increase for epithelial to mesenchymal transition (EMT) and glycolysis. Comparisons of transcripts and proteins in both CR and PD were tightly correlated (Spearman rank 0.765). Since patient-derived cell numbers were limited (&lt;50,000), we utilized genome-scale metabolic models to infer relevant metabolic pathways by comparison to the human metabolic Atlas (HMR2). We observed high enrichment in fatty acid β-oxidation processes in different organelles related to CR TIL and tumor. Lastly, we examined if increasing fatty acid oxidation in TILs might enable their increased survival in metabolically replete environments. These results show that the fatty acid metabolism pathway is associated with the ACT efficacy, and it can contribute to improving treatment.
Paradoxical pro-tumor functions of dendritic cells on colorectal cancer
Authors: Huang Hsin-I , Hammer Gianna 
Published: January 2023

Abstract
The immune system’s ability to eradicate cancer requires coordination between innate and adaptive immune cells. No other cell type bridges innate and adaptive immunity better than the dendritic cell (DC). For this reason, DCs, specifically the Batf3-dependent DC subset, are thought to be key to anti-tumor immunity. Although the anti-tumor functions of Batf3-dependent DCs have been well described in skin cancer, whether these DCs have similar anti-tumor functions in other cancers is unknown. Here we investigated the roles of Batf3-dependent DCs in colon cancer and paradoxically found that these DCs actually promote colon tumorigenesis. Thus, in a spontaneous colorectal cancer mouse model, loss of Batf3-dependent DCs resulted in decreased colon tumor number. Although Batf3-dependent DCs are best known for expanding IFNγ-producing T cells, in colon tumors, we surprisingly found that Batf3-dependent DCs were required to expand tumor-infiltrating γδT cells producing IL-17. IL-17 is known to promote colon tumorigenesis and our findings pinpoint IL-17-producing γδT cells as a key source of IL-17 for tumor growth, and surprisingly describe a requirement for Batf3-dependent DCs to expand an IL-17-producing T cell population. In addition, we identified tumor-infiltrating Batf3-dependent DCs uniquely upregulate high levels of PD-L1as a second mode of action to promote colon cancer. PD-L1 upregulation and high PD-L1 expression on Batf3-dependent DCs was specific within the tumor microenvironment. Collectively, these paradoxical pro-tumorigenic functions of Batf3-dependent DCs suggest that these DCs are not universally anti-tumor and that targeted therapies that modulate Batf3-dependent DCs could benefit colon cancer patients.
The immune suppressive role of DC-derived IL-27 in hepatocellular carcinoma via the modulation of gut microbiota
Authors: Liu Haiyan , Liu Yonghao , Song Yuan 
Published: January 2023

Abstract
IL-27 can drive the expression of co-inhibitory receptors on T cells, such as PD-1, TIM-3 and podoplanin, which may inhibit T cell functions during chronic inflammation and tumor development. However, the role of IL-27 in regulating anti-tumor immunity is still controversial. Here, we identified a tumor-promoting role of DC-derived IL-27 in murine hepatocellular carcinoma models. IL-27-p28 conditional knockout (KO) in DCs had significantly reduced tumor growth compared with the wild type (WT) controls. The numbers of T cells and NK cells were increased in the tumor microenvironment and the productions of IFN-gamma and TNF-alpha in T cells and NK cells were also enhanced in IL-27 KO mice compared with the WT mice. Moreover, deficiency of IL-27 in DCs did not affect the expression of co-inhibitory receptors on T cells from naïve mice, but reduced the expression level of PD-1 on tumor infiltrating T cells. Interestingly, when WT mice received fecal microbiota transplant from IL-27 KO mice, the tumor growth was significantly reduced. Therefore, DC-derived IL-27 may regulate anti-tumor immune response through the modulation of gut microbiota. Furthermore, we discovered that the commensal bacteria, bifidobacterium and akkermansia, were increased in IL-27 deficient tumor-bearing mice compared with the WT controls. These two strains were found to be increased during anti-PD-L1 or anti-PD-1 antibody therapy and may promote anti-tumor immunity. Thus, our results demonstrated that DC-derived IL-27 could promote tumor growth by suppressing anti-tumor immune responses possibly through the modulation of gut microbiota.
Dendritic cells dysfunction in tumor-draining lymph nodes
Authors: Bandola-Simon Joanna Marta , Roche Paul A 
Published: January 2023

Abstract
An effective anti-tumor response requires activation of both CD4 helper T cells (through MHC class II) and CD8 cytotoxic T cells (through MHC class I). Activation of tumor-specific CD4 and CD8 T cells depends on tumor antigen presentation by DCs. However, it is known that the tumor microenvironment largely suppresses immune responses. The focus of our study is to characterize the functional status of DCs in tumor-draining lymph nodes and to identify the mechanisms that lead to defective MHC class II presentation of tumor antigens. We injected either B16-F10 melanoma or MB49 bladder cancer adenocarcinoma cells into the flank of mice and isolated DCs from both tumor-draining lymph node (TDLN) and non-draining lymph nodes (NDLN) as our model system. Cross-presentation of OVA to OVA-specific CD8 T cells was not suppressed in TDLN in comparison to the NDLN either in vivo and ex vivo. However, both in vivo and ex vivo CD4 T cell activation by DCs from TDLNs was defective. Subset analysis revealed that both migratory and resident CD8-positive and CD8-negative DCs in TDLN show a less mature phenotype as compared to their counterparts isolated from NDLN. Examination of antigen uptake and processing also showed defects in DCs in TDLN. RNA-seq revealed many differences in gene expression between both resident and migratory DCs isolated from TDLN and NDLN. Characterized phenotypical and functional differences, confirmed by gene expression studies, may explain the dysfunction of DCs in tumor-bearing mice. Further analysis should enable moving towards “correcting” DC defects that would allow proper CD4 T cell priming to provide help to anti-tumor CD8 T cells.
CDDO-Me Redirects Macrophage Activation
Authors: Santiesteban Gretel Maria Torres , Shabaneh Tamer , Bhandari Rajan , Liby Karen , Turk Mary Jo , Pioli Patricia 
Published: January 2023

Abstract
Melanoma tumors are highly immunogenic, making them an attractive target for immunotherapy. However, many patients do not mount robust clinical responses to targeted therapies, which is attributable, at least in part, to suppression of immune responses by tumor-associated macrophages (TAMs) in the tumor microenvironment (TME). Using a human in vitro culture system, we now show that the synthetic triterpenoid CDDO-Me enhances immune activation by reprogramming macrophages from immuno-suppressive to immuno-stimulatory. CDDO-Me treatment inhibits surface marker expression of CD16 and CD163 and significantly reduces production of CCL2 and IL-6 in macrophages. These studies also demonstrate that CDDO-Me effects on TAM activation are enhanced when macrophages are tri-cultured with autologous activated T cells and BRAFV600Emutant cells. Because CDDO-Me dramatically attenuates secretion of CCL2, we hypothesize that CDDO-Me may enhance melanoma patient response to additional immunotherapies, including BRAF inhibition. Approximately 50% of advanced melanomas harbor activating BRAFV600Emutations, and BRAF inhibitors have consistently shown anti-tumor responses in patients with BRAFV600Emutant melanoma. However, the duration of the response has been limited due to the development of acquired resistance, which occurs because of myeloid recruitment driven by CCL2. Current studies are focused on assessing the efficacy of combination CDDO-Me/BRAF inhibition. Collectively, these studies suggest that the redirection of immuno-suppressive myeloid cell activation may provide both a direct means of inhibiting melanoma growth and may enhance the efficacy of additional targeted immunotherapies.
The effects of a CRISPR/Cas9 IL-6 knockout in 4T1 mammary carcinoma cells on myeloid-derived suppressor cells (MDSCs) and Th17/Th22 cells
Authors: Rase Viva Jeanne , Haughian James M , Pullen Nicholas A 
Published: January 2023

Abstract
The 4T1 mouse mammary carcinoma cell line is syngeneic to Balb/c mice and is an effective tumor model to study the interactions between the immune system and metastatic breast cancer. The pro-tumor effects that myeloid-derived suppressor cell (MDSC) populations have on CD8+ T cells and Tregs are well known, but comparably less attention has been directed at the relationship between MDSCs and Type III immune Th17/Th22 cells in breast cancer. MDSCs and Th17/Th22 cells have been linked in other inflammatory settings, so in this study, we investigated MDSC, Th17 and Th22 cell interactions in 4T1 tumors, and the role of tumor-derived IL-6 in these interactions. IL-6 plays a key role in the induction of MDSCs under pathological conditions, as well as fate determining in Th maturation. Thus, the removal of IL-6 from 4T1 tumors allowed us to better characterize the source of these interactions. Populations of Monocytic (M)-MDSCs (CD11b+Ly-6G−Ly-6Chi), Polymorphonuclear (PMN)-MDSCs (CD11b+Ly-6G+Ly-6Clow), Th17 (CD3+CD4+RORγt+IFN-γ−IL-17A+IL-22−) and Th22 (CD3+CD4+IFN-γ−RORγt−IL-17A−IL-22+) cells were identified in peripheral blood, spleen, tumor and bone marrow (BM) of mice bearing wild-type and IL-6 knockout 4T1 tumors. Finally, we isolated 4T1 induced MDSCs (CD11b+Gr-1+)and cocultured them with healthy donors to investigate Th17 and Th22 population expansion. Our findings provide further insight into the interactions among MDSCs, Th17 and Th22 cells in a murine cancer model, which may inform immunotherapy efficacy and outcomes in human breast cancer.
Myeloid derived suppressor cells promote ovarian cancer progression in a murine model of ovarian cancer
Authors: Rattan Ramandeep , Sakr Sharif , Dar Sajad , Buekars Thomas , Morris Robert , Munkarah Adnan , Giri Shailendra 
Published: January 2023

Abstract
Myeloid Derived Suppressor Cells (MDSCs) are a heterogeneous population of immature myeloid cells that create an immunosuppressive environment by inhibiting T-cell function. MDSCs also promote angiogenesis, tumor invasion, and metastasis. Increased MDSCs in epithelial ovarian cancer (EOC) are been associated with poor prognosis. The aim of our study was to investigate if MDSCs can influence EOC progression and if their depletion would improve immune response and restrict EOC growth. The intraperitoneal ID8 syngeneic mouse EOC model in B6 mice was used. ID8 tumor bearing mice exhibited significantly higher levels of MDSCs (CD11b+Gr1+) in bone marrow, blood, ascites and tumor compared to non-tumor control groups, including both sub-populations; granulocytic (CD11b+Ly6G+Ly6Clo) and monocytic (CD11b+Ly6G−Ly6Chi). Depleting MDSCs either by anti-Gr1 (Ly6G/Ly6C mAb) monoclonal antibody or gemcitabine treatment significantly slowed EOC progression, as reflected by decreased ascites and reduced tumor burden at the peritoneum, bowel, liver, spleen, and kidney. This was accompanied with an increased number of IFNγ-producing CD4 and CD8 T-cells. Adoptive transfer of MDSCs (Gr1hiLy6G+) enhanced ID8 tumor progression, further supporting their pathogenic role in EOC. Moreover, depletion of MDSCs enhanced the effects of anti-PD-1 immunotherapy in the ID8 EOC mouse model. Overall, we report here that depleting MDSCs improves the antitumor immune response, restricts EOC progression, and enhances tumor responsiveness to immunotherapy. Thus, targeting MDSCs is a potential therapeutic modality in EOC that may be beneficial in combination with other immunotherapies.
ST8Sia6 Overexpression Accelerates Tumor Growth, Alters Macrophage Polarization and the Immune Response
Authors: Friedman David J , Shapiro Michael , Rajcula Matthew , McCue Shaylene , Shapiro Virginia Smith 
Published: January 2023

Abstract
T-cell targeted immunotherapy has resulted in novel treatments for multiple cancers and has been the primary focus in cancer immunotherapy. However, many patients still fail checkpoint therapies due to immune suppression mechanisms driven by cells, such as tolerogenic M2-like macrophages, which support tumor growth. ST8Sia6, a sialyltransferase, generates α2,8-linked disialic acids on O-linked glycoproteins. Cancer patients with high ST8Sia6 expression exhibit reduced survival as compared to patients with low expression, but the mechanism is unknown. We generated MC38 and B16–F10 murine cancer cell lines that overexpress ST8Sia6. Expression of ST8Sia6 accelerated flank tumor growth in both single and dual flank injection models as compared to parental tumors. In vivo, macrophage polarization was altered in ST8Sia6 expressing tumors. At day 7, tumor associated macrophages (TAMs) from ST8Sia6 negative MC38 tumors were primarily M1-like, while TAMs from ST8Sia6 expressing MC38 tumors were predominately M2-like. Tumor growth and M2-like polarization was dependent Siglec-E. Siglec-E present on innate cells bind α2,8-linked disialic acids generated by ST8Sia6 and suppress immune activation. Therefore, ST8Sia6 overexpressing tumors engage Siglec-E, suppressing the immune response and increasing tumor growth. ST8Sia6 overexpression in a spontaneous murine colon cancer model decreased survival from over 6 months to 2–3 months. Therefore, ST8Sia6 expression in tumors may promote tumor growth by promoting tolerogenic immune responses.
A mixed phenotype of tumor-associated macrophages and neutrophils is evident within hours of murine mammary carcinoma delivery
Authors: Kurt Robert A , Zhang Chenyu , Schwartz Jennifer , Lauricella Amanda , Adusei Matthew , DeBenedetto Matthew , Newsom-Stewart Catherine 
Published: January 2023

Abstract
Most of what we know about tumor associated macrophages (TAM) and tumor associated neutrophils (TAN) comes from analysis of these cells isolated from solid tumors, or the periphery of humans or mice with solid tumors. These studies have largely led to a model in which tumor progression over time creates an environment that leads to the generation of pro-tumor TAM and TAN. If this model is correct then one may expect TAM and TAN that arrive early at tumor sites to largely exhibit an anti-tumor phenotype. In order to address this hypothesis we used a gelatin sponge model to capture TAM and TAN well before a solid tumor was palpable. Not surprisingly, the immune response during the first 72 hours was predominated by Ly6G+ and F4/80+ cells. While TNF-a was the most predominant cytokine produced by these cells, TGF-b was also produced at appreciable levels, and TAM found at the site of the most immunogenic of the tumors exhibited a significant increase in anti-tumor (TNF-a) as well as immune suppressive (arginase, TGF-b) cytokines between 24 to 72 hours post tumor injection. In addition, while there were more ROS+ TAN at the tumor sites compared to control sites at 72 hours, the amount of ROS in the cells at the tumor sites was lower than the amount of ROS in the cells at the control sites. Finally, we looked at metabolism because increased oxidative phosphorylation has been correlated with an immune suppressive phenotype. We found that 72 hours after tumor delivery there was already a significant shift toward oxidative phosphorylation in the TAM. Collectively, these data support the contention that an immune suppressive environment is well underway at the early stages of tumor growth; before a tumor is even palpable.
Selective IL-2 receptor β (CD122) targeting improves αPD-L1 immunotherapy in a metastatic bladder cancer model
Authors: Reyes Ryan Michael , Zhang Deyi , Deng Yilun , Mukherjee Neelam , Ji Niannian , Gupta Harshita B , Svatek Robert S , Curiel Tyler J 
Published: January 2023

Abstract
αPD-L1 is an FDA-approved bladder cancer (BC) immunotherapy but is effective in ≤30% of cases. We tested conjugates of αIL-2 antibody + IL-2 (IL-2c) that block IL-2 from binding high-affinity IL-2Rα (CD25) for preferential IL-2Rβ(CD122) binding. CD25 and CD122 are preferred for IL-2 capture by regulatory T cells and anti-tumor effector T cells (Teff), respectively, allowing IL-2c to target Teff preferentially. Orthotopic, intravesical (in bladder) MB49 BC produces PD-L1+BC tumors in syngeneic BL6 mice. αPD-L1 or IL-2c treated intravesical, but not lung metastatic MB49 produced by intravenous injection. Still, in metastatic BC, αPD-L1 + IL-2c (combo) reduced lung metastases and extended survival. Preliminary data found combo treatment efficacy was better in lung versus bladder. In subcutaneous (SQ) B16 melanoma, combo was better than single agents, and increased CD8+CXCR5+TCF-1+Tim-3−PD-1+T stem cells (CXCR5+SC, see X. Zhang poster) vs. single agents. Neither αPD-L1 nor combo increased CXCR5+SC in MB49 in bladder or lung metastases, suggesting a novel treatment mechanism. To test the impact of tumor PD-L1 on treatment efficacy, we made PD-L1KOMB49, but it did not grow in bladder or SQ in wild type mice, but grew similar to control MB49 in immunodeficient NSG mice SQ, suggesting tumor PD-L1 microenvironment-specific effects. PD-L1KOB16 grew well SQ in wild type mice, excluding a PD-L1-specific defect. Thus, tumor PD-L1 differs in immune evasion in a tumor-dependent manner. Selective IL-2 targeting to CD122 improves αPD-L1 treatment of metastatic BC. Mechanisms differ from melanoma, which could be due to tumor, PD-L1, or microenvironment effects. We are assessing mechanisms and αPD-1 treatment effects (also FDA-approved for BC).
Enhanced antitumor effects by combining IL-12 gene electrotransfer with anti-PD1 in preclinical metastatic cancer model
Authors: Shi Guilan , Lundberg Cathryn , Scott Megan , Edelblute Chelsea , Heller Richard 
Published: January 2023

Abstract
The survival and expansion of effector cytotoxic T lymphocytes (CTLs) during an immunological response are critical for the successful elimination of cancer cells. Among the factors that regulate the immune response, interleukin-12 (IL-12), is known to stimulate growth and survival of T cells. Major differences exist between these lymphokines in their capacity to act on various T-cell types such as CD4+ or CD8+ and effector versus memory T lymphocytes. PD1 expression can have a negative effect on effector or memory T-cell populations causing anergy or cell death. Limited information is available regarding the function of combining IL-12 gene electrotransfer (GET) with anti-PD1 in preclinical metastatic cancer models. Here, we report that IL-12 (GET) induced expansion of effector CD8 CTLs. Combining IL-12 GET with anti-PD1 i.p. injection resulted in long-term survival of CD8+PD1− T cells. Most significantly, the effector cytolytic activity of CTLs were expanded and maintained for up to 60 days, indicating that these cells do not differentiate into a memory functional phenotype. In a two site metastatic model, B16F10 melanoma cells expressing luciferase were injected via intraperitoneal route and B16.F10 cells (no luciferase) were injected subcutaneously. Results on day 60 using an In Vivo Imaging System (IVIS) illuminated a lower flux signal in intraperitoneal metastases. Simultaneously, the primary subcutaneous tumors completely regressed in 80% of these mice. The expression of CD44+ CD62L− increased with time, which was detected on CD8 CTLs, suggesting that the memory response was still active at 120 days. The present findings have practical implications for the combination of IL-12 gene electrotransfer with anti-PD1.
Isoform specific anti-TGFβ therapy enhances antitumor efficacy in mouse models of cancer
Authors: Budhu Sadna , Gupta Aditi , Giese Rachel , van Snick Jacques , Uyttenhove Catherine , Ritter Gerd , Wolchok Jedd D , Merghoub Taha 
Published: January 2023

Abstract
TGFβ is a pleotropic cytokine, which has emerged as a potential target in cancer treatment due to its dual role in tumorigenesis and homeostasis. There are three isoforms of TGFβ (TGFβ1, TGFβ2 and TGFβ3), which are secreted by immune and nonimmune cells as an inactive latent complex. Depending on the local context and players, TGFβ can adopt opposing roles in carcinogensis and in modulating the immune system. However, the expression of TGFβ isoforms within the tumor microenvironment and isoform specific inhibition remains to be investigated. The main source of TGFβ isoforms in the tumor microenvironment of B16 melanoma are infiltrating immune cells, with TGFβ1 and TGFβ3 being highly expressed on myeloid and dendritic cells. The CD45− population from B16 tumors demonstrated a lower expression of both TGFβ isoforms. Compared to untreated control animals, anti-TGFβ3 therapy resulted in the greatest delay in B16 tumor growth, followed by anti-TGFβ1 therapy and pan-TGFβ blockade. However, none of the therapies resulted in improved overall survival. Similar results were achieved in a 4T1 breast model. T cell functional assays demonstrated that anti-TGFβ3 resulted in CD8+ T cells with greater cytolytic ability as they showed higher granzyme B expression and killing against B16 cells when plated ex-vivo. Anti-TGFβ1 treatment resulted in greater interferon-γ production by CD8+ T cells, suggesting an increase in antigen-specificity. Isoform specific TGFβ inhibition in combination with immune checkpoint blockade demonstrated improved tumor control and survival. This provides rationale for the use of anti-TGFβ therapy in stroma poor tumors, such as melanoma, and for its potential to enhance the effectiveness of existing therapies.