bluuebunny/crossref_metadata_2025_split huggingface dataset:
Super2-expressing CAR T cells generate improved anti-tumor responses against solid tumors
Authors: Brog Rachel A , Kumar Prathna , Sentman Charles , Huang Yina 
Published: January 2023

Abstract
Chimeric antigen receptors (CAR) augment tumor specific immune responses by redirecting all T cells towards recognition of tumor antigen. Although CAR T cell treatment shows promising therapeutic effects in leukemia patients, current CAR therapies are inadequate against solid tumors, in part from exclusion or functional exhaustion in the suppressive microenvironment. Exogenously administered Interleukin 2 (IL-2) improves efficacy and expansion of CAR T cell therapy. However, IL-2 administration is limited by high toxicity and low half-life. Super2 is an engineered IL-2 variant that binds with high affinity to the beta and gamma chains of the IL-2 receptor even without IL-2R alpha chain expression. We found that Super2 leads to increased phosphorylation of STAT-5 and more effectively activates lymphocyte populations compared to endogenous IL-2 in vitro. We hypothesized that engineering CAR T cells to express Super2 will improve their functionality within the tumor microenvironment. CAR T cell-intrinsic Super2 may also increase activation of the endogenous tumor-reactive T cells resulting in enhanced tumor control and the establishment of long-term memory even after loss of CAR T cells. To test this hypothesis, we designed CAR T cells that produce Super2 constitutively or in response to CAR stimulation. We found that Super2-expressing CAR T cells increased tumor infiltrating leukocytes (TILs) compared to CAR T cell treatment alone in immune competent murine models. Additionally, Super2-expressing CAR T cells delayed B16F10 melanoma growth in comparison to CAR T cell alone. We conclude that Super2-expressing CAR T cells provide improved protection against solid tumors.
Nutrient partitioning in the tumor microenvironment and FDG-PET imaging
Authors: Madden Matthew Z , Reinfeld Bradley I , Wolf Melissa M , Cohen Allison S , Manning H Charles , Rathmell W Kimryn , Rathmell Jeffrey C 
Published: January 2023

Abstract
The tumor microenvironment is composed of multiple cell types, including malignant cancer cells and tumor-infiltrating leukocytes (TIL). A possible mechanism of immunosuppression in the tumor microenvironment (TME) is cancer cells outcompeting anti-cancer TIL for nutrients such as glucose. 18F-deoxyglucose (FDG) positron emission tomography (PET) imaging is a staple of diagnosing and monitoring many types of cancer and is based on the Warburg model in which cancer cells within the tumor utilize glucose for growth and proliferation. Here, we use magnetic bead sorting to fractionate FDG-avid murine tumors and measure tumor cell-specific glucose uptake. We find that CD45+ immune cells are more FDG-avid than CD45− cancer cells. We further fractionate immune cell subsets into CD4/8+ T cells and CD11b+ myeloid cells to demonstrate that TIL T cells take up more glucose than CD45− cancer cells and resting splenic T cells. Strikingly, CD11b+ myeloid cells are the most FDG-avid cells in the TME. In MC38 colorectal cancer tumors, we show that CD11b+ F4/80 hi macrophages have high glucose uptake, and by extracellular flux analysis demonstrate higher metabolic activity than tumor T cells and CD45− cancer cells. Intriguingly, while glucose uptake is low in CD45− cancer cells, 18F-glutamine uptake is higher in cancer cells than immune cells. Our results illustrate a novel approach to measuring nutrient uptake in the TME and suggest that TIL are not starved of nutrients in the TME. Future work will determine the effects of immunotherapy and metabolism-targeted therapeutics on cell-specific glucose and glutamine uptake in the TME, as well as determine the immune contribution to cancer PET scans in the context of immunotherapy response.
Arid5a orchestrates immune evasion of pancreatic cancer via the Ido1-–mediated tryptophan metabolism
Authors: Parajuli Gyanu , Hashimoto Shigeru , Kishimoto Tadamitsu 
Published: January 2023

Abstract
Although pancreatic ductal adenocarcinoma (PDAC) is known to be “immunologically cold” tumors, in which the presence of immune cells and their activity are limited by the tumor microenvironment, the immunological nature of this cancer is still largely unknown. AT-rich interaction domain-containing protein 5a (Arid5a) was identified as an RNA-binding protein that plays significant roles in the development of inflammatory diseases via the augmentation of IL-6 signaling. However, the role of Arid5a in cancer remains elusive. To clarify the role of Arid5a in cancer, we used the well-established mouse PDAC model KPC cells, bearing KRAS/TP53 mutations. We found that although KPC cells lacking Arid5a (Arid5a-KO) had similar growth rates as wild-type (WT) KPC cells when subcutaneously inoculated into immunodeficient mice, Arid5a-KO cells had remarkably impaired their growth in immunocompetent mice. Here, we show that Arid5a promotes immunosuppressive tumor microenvironment by inducing the infiltration of regulatory T cells and granulocytic myeloid-derived suppressor cells; and inhibiting the activation of CD8+T lymphocytes. Interestingly, we observed that Arid5a-KO KPC cells had significantly lower expression of indoleamine 2,3-dioxygenase 1 (Ido1) compared with WT KPC cells. Mechanistically, we identified that Arid5a stabilizes Ido1 mRNA through binding to its 3′-untranslated region, and augments the production of kynurenine, which is an immunoregulatory metabolite of Ido1-mediated tryptophan catabolism. Our findings indicate Arid5a as a crucial molecule involved in the immune evasion of PDAC which can be exploited for immunotherapy in malignant pancreatic cancer.
Agonistic-CD40 therapy alters the spatial localization of immune cells in the tumor microenvironment: implications for therapeutic responsiveness
Authors: Maltez Vivien , Byrne Katelyn , Germain Ronald N 
Published: January 2023

Abstract
Despite major advances in immune checkpoint blockade therapies, tumor therapeutic responsiveness remains difficult to predict. This is especially true for pancreatic ductal adenocarcinoma, where total T cell numbers in the tumor proper do not predict responses to immune checkpoint blockade. Here, we use a novel congenic KPC pancreatic mouse model-derived tumor clone library that recapitulates the spectrum of tumor heterogeneity seen in patients, enabling us to perform head-to-head comparisons between therapeutically responsive and refractory tumors. Previous flow cytometry-based work with these clones revealed that tumor-derived CXCL1 is a major determinant in establishing a therapy-refractory environment, and that CXCL1 ablation enables therapeutic responses. Strikingly, we found that refractory tumors contain numerous myeloid clusters not present in responsive tumors, and that CXCL1 ablation abolishes this cluster formation. We hypothesized that these immunosuppressive myeloid clusters could critically hamper the spatial distribution of tumor infiltrating immune cells. In support of this, the spatial organization of multiple immune cell subsets, including regulatory T cells, dendritic cells, and CD8 T cells, is vastly different between responsive and refractory tumors. Upon therapeutic intervention, the spatial organization is altered in both tumor environments, with responsive tumors developing clusters of activated dendritic cells and T cell subsets, and refractory tumors developing an increased number of immunosuppressive myeloid clusters. These data suggest that therapeutic efficacy is determined by foci of local immune activity, a phenomenon that could function as a positive diagnostic biomarker.
Antibody-mediated Recruitment of Viral-specific Effector Cells via Recombinant Antibody-MHC T cells Engagers
Authors: Cohen Maya , Efimov Vera , Reiter Yoram 
Published: January 2023

Abstract
Our lab has developed a novel T cell engager (TCE) whereby a recombinant antibody fragment specific for a tumor-associated antigen (TAA) of choice is genetically fused to HLA-A2/peptide complex that presents a viral T cell epitope. The HLA-A2/peptide complex is designed to engage the TCR of a CD8+ T cell while the scFV or Fab redirects specificity towards a TAA, thus recruiting potent antiviral memory CTLs to attack tumor cells. We used the highly immunogenic cytomegalovirus (CMV) pp65-derived peptide as a target peptide for CTLs, taking into consideration the CMV prevalence of seropositive humans. Mesothelin was used as a TAA.
We monitored the binding of TCEs to target cells demonstrating that their binding is dependent exclusively on the interaction of the targeting domain (scFv/Fab) of the fusion molecule with mesothelin, and is dose-dependent. Potency of TCEs was determined in vitro using co-cultures of CMV-specific CTLs and mesothelin-expressing human tumor cell lines. The TCEs specifically and potently mediated the killing of mesothelin-positive cells in TCE-dose-dependent matter. Moreover, a significant increase in CD25 expression and dose-dependent secretion of IL2 and IFNγ were observed in CTLs exposed to mesothelin-positive cells.
The results represent a powerful new approach for immunotherapy. The MHC-based TCEs can mediate specific and efficient recruitment of viral-specific CTLs to kill tumor cells, adding an approach by which specific population of T cells can be engaged and recruited to the tumor site in a controllable manner in contrast to current CD3-based T cells engagers that cannot selectively target the specificity of interest. Such treatments can be very potent but with decreased toxicities involved.
Metronomic low dose cyclophosphamide in combination with a DNA methyltransferase inhibitor limits tumor growth in murine triple negative breast cancer
Authors: Camarena Carmen , Smith Timothy M , Bear Harry , Martin Rebecca 
Published: January 2023

Abstract
Many breast cancers are treated with therapies that target herceptin, estrogen, or progesterone receptors; however, for triple-negative breast cancers (TNBC) that lack these receptors, treatment options are limited, and prognosis is often unfavorable. The goal of this study is to design a therapeutic intervention that can elicit an effective immune response against the tumor and instill immunological memory to eradicate primary and metastatic lesions. Guadecitabine (Guad) is a second-generation DNA methyltransferase inhibitor (DMNTi) that has been reported to have several antitumor properties such as increased antigenicity and depletion of myeloid-derived suppressor cells (MDSC’s). Cyclophosphamide is a FDA approved chemotherapy that has been shown, when given as a low-dose treatment, to selectively deplete regulatory T-cells (T-regs). Both MDSCs and T-regs suppress antitumor immunity. We hypothesize that the combination of Guad and Cyp will synergize and promote anticancer immunity through increased expression of de novo tumor antigens and depletion of MDSCs and T-regs in a low-dose setting. To test this hypothesis, BALB/c mice were challenged with murine TNBC 4T1 tumor cells and the 4T1-bearing mice were administered low-dose Guad and Cyp daily for ten consecutive days. This experiment showed a degree of synergy between Guad and Cyp with the dual therapy reducing tumor burden to a greater extent than either monotherapy.
All-trans Retinoic Acid (atRA) promotes tumor resistence to NK cells by downregulating tumor B7-H6 expression
Authors: Cao Guoshuai , Sun Rui , Tian Zhigang 
Published: January 2023

Abstract
The interaction between the activating receptor NKp30 and its major tumor ligand B7-H6 is important for NK cell mediated tumor rejection while the regulation of B7-H6 by tumor therapeutics remains largely unknown. In this study, we investigated the regulation of B7-H6 by all-trans Retinoic acid (atRA), which is a terminal differentiation inducer of tumor cells and in use for tumor therapy. We found that B7-H6 expression on tumor cell suface was downregulated upon atRA treatment. We further found that atRA downregulated B7-H6 at the mRNA level. Epgenetic modifications such as DNA methylation and HDAC was not responsible as inhibitors of these pathways did not restore the B7-H6 mRNA expression. Instead, atRA downregulated c-myc and inhibited the transcription of B7-H6. NK92 and NKG cells mediated U937 cell killing was mainly dependent on the NKp30-B7-H6 interaction. Thus atRA treatment dampened U937 lysis by NK92 and NKG cells. Together, our data showed that atRA treatment downregulated B7-H6 expression and promoted tumor cell resistence to NK cells.
Paradoxical Role of MLK3-CTLA-4 axis in Triple-negative Breast Cancer and CD8+ T cells
Authors: Kumar Sandeep , Singh Sunil Kumar , Viswakarma Navin , Sondarva Gautam , Nair Rakesh Sathish , Thatcher Gregory , Rana Basabi , Rana Ajay 
Published: January 2023

Abstract
Mixed Lineage Kinase 3 (MLK3), a MAP3K family member, is an upstream regulator of MAP kinases and reported to mediate cytotoxic drug-induced breast cancer cell death, suggesting that MLK3 could serve as a therapeutic target in breast cancer. However, the function of MLK3 in host immunity and Triple-negative Breast Cancer (TNBC) are not known. The small-molecule inhibitor of MLK3, URMC-099, was able to induce caspase activity and promote apoptosis in TNBC cells, both in vivo and in vitro settings. The cytokine levels in TNBC xenograft tumors were either decreased or unchanged except for a significant increase in IL-1β level upon MLK3 inhibition. However, most of the Th1/Th2/Th17 cytokines were found to be up-regulated in the serum of tumor-bearing mice. The expression of T cell activation marker, CD69 on splenic, and tumor-infiltrating CD8+ T cells was increased upon inhibition of MLK3. Furthermore, the number of effector CD8+ T cells was also increased upon inhibition of MLK3 in tumor-bearing mice. We also observed that proteins, implicated in CD8+ T cell exhaustion, TIM3, LAG3, NFATc1, and CTLA-4 were up-regulated on CD8+ T cells upon MLK3 inhibition. Additionally, genetic loss/pharmacological inhibition of MLK3 increased expression of CTLA-4 on TNBC and CD8+ T cells via NFATc1. Paradoxically, the cellular proliferation of TNBC cells, expressing CTLA-4 was up-regulated, however it was opposite in CD8+ T cells. Most importantly, the combined blockade of MLK3 and CTLA-4 increased tumor infiltration of cytotoxic CD8+ T cells and correspondingly decreased the cellular proliferation and survival of tumor cells. Collectively, these results suggest that combined blocked of MLK3 and CTLA-4 could serve as a novel immunotherapy approach for TNBC.
Utilization of genetically diverse mouse model identifies genetic loci that correlate to response to immune checkpoint inhibitors to B16F0 melanoma
Authors: Hackett Justin , McCasland Jennifer , Wiesend Wendy , Dyson Greg , Muñiz Maria , Gibson Heather 
Published: January 2023

Abstract
Historically metastatic melanoma has been associated with poor prognosis. The advent of immune checkpoint inhibitors targeting CTLA-4 and PD-1 have greatly improved patient survival in metastatic melanoma, but a significant proportion of patients fail to respond to therapy. To explore the effect of genetic background on immune checkpoint inhibitor efficacy, we have developed a tumor immunotherapy model using genetically heterogeneous Diversity Outbred (DO) mice and the C57BL/6 syngeneic B16F0 melanoma model. To implement genetic diversity, DO mice were crossed with C57BL/6 to generate (C57BL/6xDO)F1 mice. Untreated (C57BL/6xDO)F1 mice (n=34) reliably develop B16F0 tumors after subcutaneous inoculation, with some variation in tumor latency. To test the role of genetics in response to ICI, (C57BL/6xDO)F1 mice (n=142) were treated with combined anti-PD1/anti-CTLA-4 on days 3, 6, and 10 after inoculation with 2×105 B16F0 cells. Mice receiving therapy show wide variation in tumor development of up to 65 days (mean 20.86 +/− 11.04, CV 52.94%), with 19 never developing tumor by day 88. Re-challenge of tumor free mice confirmed adaptive response to tumor. Preliminary Genome Wide Association shows suggestive associations on chromosomes 8, 12, and 13. The locus on chromosome 13 contains a family of related genes with known function in autoimmune conditions and expression in several immune cell types. QTL effects show the NZO genotype is a positive driver at this locus and C57BL/6 is a negative driver, reflecting known ICI resistance in the C57BL/6 strain. These results demonstrate host genetic background significantly contributes to response to ICI and identifies genomic loci associating with primary resistance. NIH R37 CA220482.
Redirecting Immune Activation in the Tumor Microenvironment of Estrogen Receptor Negative Breast Cancer
Authors: Pioli Patricia Ann , Ball Michael S. , Bhandari Rajan , Santiesteban Gretel Maria Torres , Martyanov Viktor , Eltanbouly Mohamed , Whitfield Michael L , Liby Karen 
Published: January 2023

Abstract
The tumor microenvironment (TME) is an essential contributor to the development and progression of malignancy. Within the TME, tumor associated macrophages (TAMs) mediate angiogenesis, metastasis, and immune suppression, which inhibits infiltration of tumor-specific cytotoxic CD8+ T cells. In previous work, we demonstrated that the synthetic triterpenoid CDDO-methyl ester (CDDO-Me) converts breast TAMs from tumor-promoting to tumor-inhibiting in vitro. We show now for the first time that CDDO-Me remodels the breast TME, redirecting TAM activation and T cell tumor infiltration in vivo. We demonstrate that CDDO-Me significantly attenuates IL-10 and VEGF expression but stimulates TNF production, and reduces surface expression of CD206 and CD115, markers of immunosuppressive TAMs. CDDO-Me treatment redirects the TAM transcriptional profile, inducing signaling pathways associated with immune stimulation, and inhibits TAM tumor infiltration, consistent with decreased expression of CCL2. In CDDO-Me-treated mice, both the absolute number and proportion of CD4+ T cells were reduced, while the proportion of CD8+ T cells was significantly increased in tumors and spleen. Moreover, mice fed CDDO-Me demonstrated significant reductions in numbers of tumor CD4+Foxp3+ regulatory T cells. These results demonstrate for the first time that CDDO-Me relieves immunosuppression in the breast TME and unleashes host adaptive anti-tumor immunity.
Fine Needle Aspiration Based Immune Organoids Recapitulate TME in clear cell Renal Cell Carcinoma (ccRCC)
Authors: Wolf Melissa , Madden Matthew Z , Hongo Rachel , Rathmell W. Kimryn , Weiss Vivian L 
Published: January 2023

Abstract
Replication of the tumor microenvironment (TME) is a major limitation in organoid development as a model system for cancer research. Studies aimed at understanding the TME are typically performed in an oversimplified 2D cell culture system, or utilizing complex mouse models. Not only are in vivo mouse studies time consuming and costly, significant differences in the mouse immune system and TME may hinder applicability of the results in a translational paradigm. Here, we show that human specimens obtained via fine needle aspiration (FNA) from cancerous clear cell renal cell (ccRCC) tumors can grow and be maintained as immune organoids harboring a diverse lymphoid and myeloid population in a matrigel-based culture model. This system provides several advantages over cocultured cell lines with primary immune cells. In our FNA based method, the immune cell population is obtained directly from the TME, reflecting a realistic representation of tumor-associated exhaustive and/or regulatory immune cells. In addition, these studies provide a framework for drug development and allow for the evaluation of a 3D organization of tumor cells with immune cells. Ultimately, there is a translational gap between 2D cell culture and animal model testing; therefore, evaluation of the 3D organization of tumor cells within an ideal TME could recapitulate physiological cancer processes.
IL-6 Promotes Cell Growth and Is Associated With Poor Prognosis In Patients With Oral Cancer
Authors: Chia Jean-San , Wei Ling-Ying 
Published: January 2023

Abstract
Interleukin-6 (IL-6) is a pleiotropic inflammatory cytokine that regulates growth and differentiation of various types of malignant tumors, including oral cancers. Compared to healthy controls, the levels of IL-6 are elevated in sera of patients with oral cancer. In this study, the role of IL-6 in the growth regulation of oral cancer cells was evaluated.
The effects of IL-6 on cell growth were evaluated by expressing IL-6 cDNA into the human tongue cancer cell line (SAS), resulting in high-producer clones. In vivo, the effects of tumor-derived IL-6 on growth were studied by using an experimental vaccine model. Two stable clones, consisting of low-producer (IL-6LE), and high-producer (IL-6OE), were subcutaneously inoculated on the flank of each severe combined immune-deficient (SCID) mice. A total of 111 surgically treated HNSCC patients were studied. IL-6 and STAT3 levels were measured by immunohistochemical staining and correlated with clinicopathological variables and patient survival.
Treatment of recombinant IL-6 and overexpression of IL-6 by intorducing IL-6 into SAS cells significantly increased clonogenic ability and cell proliferation in vitro compared to the parental SAS cells and the vector controls. In animal model, a statistically significant difference of tumor weight was obtained between the IL-6LE and IL-6OE (*P=0.024) groups in experiment. Clinical data revealed significant relationships between IL-6 and stage*, lymph node metastasis*, recurrence**, disease-free survival* and overall survival* (P &lt; 0.01; *P &lt; 0.05).
These findings strongly support the hypothesis that IL-6 acts as a significant autocrine growth factor in vivo and in vivo oral cancers.
Role of the receptor tyrosine kinase AXL in regulating antitumor response and relationship with hypoxic stress
Authors: Mami-Chouaib Fathia , Terry Stephane , Buart Stephanie , Gausdal Gro , Angelsen Agnete , Lorens James , Chouaib Salem 
Published: January 2023

Abstract
Tumor hypoxia has long been considered as a driving force of tumor progression and to play a key role in remodeling the tumor stroma and favoring the emergence of tolerance. The receptor tyrosine kinase AXL is associated with drug resistance in several solid tumors and AXL-targeting agents are currently in clinical trials. We investigated the relationship between tumor hypoxia, AXL expression and the emergence of tumor resistant variants. Our data indicate that hypoxia promotes EMT, AXL expression and resistance to CTLs and NK in NSCLC. We have next investigated the molecular basis of AXL-induced resistance to cell-mediated cytotoxicity and demonstrated that AXL targeting resulted in an increase in target susceptibility to CTLs and NK mediated cell lysis by a mechanism involving ICAM and the NKG2D ligand (ULBP1).
We next investigated the effect of pseudo hypoxia (hypoxic stress in presence of oxygen) in clear cell renal cell carcinoma (ccRCC). In fact, in this type of cancer, Von Hippel-Lindau (VHL) mutations induce a hypoxic microenvironment under normoxic conditions. We showed that pseudo-hypoxia induced by VHL dysfunctions, promotes ccRCC resistance to immune cell-mediated cytotoxicity by a mechanism involving HIF-2α.
We then sked wheter the pseudo-hypoxia associated with VHL mutation in RCC could impact AXL expression. We obtained in vtro evidence indicating that both HIF2α and EMT associated transcription factors (ZEB1, ZEB2) play a role in AXL expression regulation. Using a cohort of 324 metastatic RCC patients treated with anti-PD1, we could demonstrate that AXL expression may interfere with the clinical outcome. The relationship between VHL dysfunction, AXL expression and patient response to anti-PD1 will be discussed.