bluuebunny/crossref_metadata_2025_split huggingface dataset:
PID1 promotes HCC development through regulating TME and inhibiting inflammatory factors induced apoptosis
Authors: Yang Jian , Xie Mengyuan , Xiang Ming 
Published: February 2023

Abstract

Objective:
PID1 (Phosphotyrosine interaction domain containing 1), an obesity related gene, was reported to induce insulin resistance and play a vital role in lipid metabolism in liver. However, its influence on HCC (hepatocelluar cancer) have not been full clarified. Here, we aims to investigate the mechanism of PID1 in favoring HCC development.

Method:
Hydrodynamic injection was applied to overexpress PID1 specifically in liver. Adaptive immunity was evaluated through flow cytometry. Lipids metabolism was determined by metabonomics. Western blotting was utilized to determine the effects of PID1 on PI3K/AKT and RAS/RAF/ERK pathway.

Result:
Overexpression of PID1 specifically in liver induced HCC development within 16 weeks. The frequency of CD4+ and CD8+ T cells decreased whereas the ratio of Gr1+CD11b+ cells increased in mice bearing PID1 plasmid hydrodynamic injection. Moreover, PID1 induced liver metabolism disorder, resulted in upregulation of TG, TC, NEFA, LDL-C as well as lactate levels, thereby involved in regulating TME (tumor microenvironment) and depressed adaptive immunity. Importantly, PID1 inhibited PI3K/AKT pathway, thus restrained phosphorylation of c-RAF on ser259 site induced by p-AKT and promoted phosphorylation of c-RAF on ser338 site, leading to activation of RAS/RAF/ERK pathway. Activated c-RAF targeted to mitochondria to decrease ROS level, which alleviated toxic effect of inflammatory factors and favored HCC cells survival.

Conclusion:
Our results demonstrated that PID1 provoked HCC development through regulating TME to depress adaptive immunity and inhibiting apoptosis induced by inflammatory factors.
Reciprocal interactions between the gut microbiome and mammary tissue microenvironment promote distant regulation of breast tumor metastasis
Authors: Feng Tzu-Yu , Azar Francesca N. , McGinty Mitchell T. , Rutkowski Melanie R. 
Published: February 2023

Abstract
Establishing commensal dysbiosis, defined as an inflammatory gut microbiome with low biodiversity, prior to breast tumor initiation enhances early dissemination of hormone-receptor positive (HR+) mammary tumor cells into the circulation, distal lymph nodes, and lungs. Here, we sought to define mammary tissue mediators of dysbiosis-induced tumor dissemination. Commensal dysbiosis increased mast cell numbers in the pre-malignant mammary tissue, with numbers remaining elevated in the presence of a tumor. During early tumor progression, increased mast cell numbers correlated with increased numbers of activated fibroblasts and collagen I protein levels in mammary tissues. Fibroblast activation and tissue remodeling are associated with enhanced breast tumor metastasis. Although mast cell degranulation is known to trigger fibrosis in other disease models, it is unknown whether mammary tissue mast cells influence dissemination of tumor cells. To investigate the role of mast cells during tumor dissemination, mice were treated with mast cell stabilizers ketotifen or cromolyn. Notably, inhibition of mast cell degranulation significantly reduced fibroblast activation and dissemination of tumor cells into the blood of dysbiotic animals. Collagen levels in mammary tissues from women diagnosed with HR+ breast cancer also correlated with mast cell abundance, suggesting mast cell-mediated fibroblast activation is relevant to human disease. Together, these data demonstrate that mast cells recruited into the pre-malignant mammary tissue in response to commensal dysbiosis orchestrate early dissemination of HR+ breast tumors through activation of tissue fibroblasts.
ST8Sia6 Overexpression Accelerates Tumorigenesis in a Siglec-E Dependent Manner Through Reshaping the Tumor Immune Microenvironment
Authors: Friedman David J , Crotts Sydney B , Shapiro Michael J , Rajcula Matthew , McCue Shaylene , Khazaie Khashayarsha , Dong Haidong , Shapiro Virginia Smith 
Published: February 2023

Abstract
Checkpoint blockade has improved patient outcome through reinvigorating T cells against cancer. Despite this success, many patients don’t respond, leaving them with few options. Tumors utilize many immunosuppressive mechanisms, including altering the function of innate immune cells. Cancer cells can promote differentiation of macrophages into tolerogenic “M2-like” macrophages, which support tumor growth. Macrophage differentiation by cancer cells is influenced by many receptors, including Siglec-E. Siglec-E binds to α2,8-linked disialic acids, generated by the enzyme ST8Sia6, on O-linked glycoproteins, and functions to inhibit activation of innate immune cells. In some tumor types, higher expression of ST8Sia6 correlates with poorer survival. Therefore, ST8Sia6 overexpressing tumors may engage Siglec-E, leading to immune suppression and accelerating tumor growth. Mice injected with ST8Sia6 overexpressing MC38 and B16-F10 tumors had increased tumor burden, reduced survival and increased M2 macrophage polarization, which suppresses the immune response. This acceleration of tumor growth was abrogated when injected into Siglec-E knockout mice. Thus, the primary function of ST8Sia6 in tumors is an extrinsic alteration of the immune response, rather than an intrinsic effect on cell growth. Similarly, in a spontaneous colon cancer model, ST8Sia6 overexpression in a murine colon cancer model accelerated the development of tumors, decreasing survival to 2.5 months instead of 6 months. The overexpression of ST8Sia6 leads to increased Arg-1 expression by colonic tumor macrophages. Therefore, ST8Sia6 expression in tumors inhibits the immune response to promote tumor growth in a Siglec-E dependent manner.
Genetic linkage analysis using Diversity Outbred mice identifies prolactin as a host-intrinsic regulator of immune checkpoint inhibitor response
Authors: Hackett Justin , Glassbrook James , Múniz Maria , Gibson Heather 
Published: February 2023

Abstract
The use of immune checkpoint inhibitors (ICI) 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. Incorporating what has been learned in autoimmune conditions, we hypothesize the host genetic background plays a significant role in determining response to immunotherapy. To implement genetic diversity, Diversity Outbred (DO) mice were crossed with C57BL/6 to generate (C57BL/6xDO)F1 mice that reliably develop B16F0 tumors after subcutaneous inoculation. (C57BL/6xDO)F1 mice (n=142) treated with combined anti-PD1/anti-CTLA-4 on days 3, 6, and 10 after inoculation show wide variation in tumor latency up to a maximum of 65 days, with 19 mice never developing tumor. Genetic linkage analysis using the R/qtl2 package shows a suggestive association within the murine prolactin family on chromosome 13. Visualization of QTL effects by DO founder identifies the NZO mouse genotype as a driver of increased tumor latency. Validation was performed using Collaborative Cross (CC) mouse lines that were either the NZO or C57BL/6 genotype within this locus. (C57BL/6xCC)F1 mice containing the NZO genotype have lower tumor burden by day 15 compared to mice with the C57BL/6 genotype, validating the locus as influential on the response to ICI. Combination of systemic prolactin treatment and ICI shows evidence of an interaction effect. Mice treated with both prolactin and ICI show lower tumor burden by day 19 versus single therapy or placebo controls. These results show that the host’s genetic background significantly contributes to response to ICI, and modulation of prolactin expression may augment ICI activity.
Tumor-intrinsic IRF5 reprograms the tumor immune microenvironment to inhibit breast cancer metastasis
Authors: Barnes Betsy J , Li Dan , Somerville Carter , Castro-Piedras Isabel , Pruitt Kevin 
Published: February 2023

Abstract
Metastasis of breast cancer and recurrence to incurable disease are leading causes of mortality. Metastasis is driven by extensive cooperation between a tumor and its microenvironment, resulting in the adaptation of molecular mechanisms that evade the immune system and enable pre-metastatic niche (PMN) formation. Little is known of the tumor-intrinsic factors that regulate these mechanisms. Here we show that expression of the transcription factor interferon regulatory factor 5 (IRF5) in breast cancer clinically correlates with tumor immunogenicity, increased T cell and M1 macrophage recruitment and prolonged survival. Conversely, loss of tumor-intrinsic Irf5 establishes a tumor microenvironment and PMN that supports metastasis. Mechanistically, Irf5 alters the composition and secretion of tumor-derived extracellular vesicles (t-dEVs). Upon whole-body pre-conditioning with t-dEVs from Irf5-positive or −negative breast cancer cells, we found increased lung metastatic colonization that replicated findings from orthotopically implanted breast cancer cells. Collectively, our findings uncover an essential role for IRF5 in breast cancer metastasis through its regulation of t-dEV and immune reprogramming.
Hypoxia profoundly alters Natural Killer cell phenotype and function: implications for immunotherapy within the solid tumor microenvironment
Authors: Arvindam Upasana Sunil , Kennedy Philippa , Ettestad Brianna , Phung Shee Kwan , Hinderlie Peter , Lim James , Miller Jeffrey S , Felices Martin 
Published: February 2023

Abstract
For Natural Killer (NK) cell-based immunotherapy to succeed against solid tumors, NK cells need to enter a highly immunosuppressive tumor microenvironment (TME) and retain functionality. The objective of this study is to evaluate how hypoxia exerts an immunosuppressive effect on NK cells. We used the novel AVATAR™ system to model oxygen levels of three key tissues that NK cells inhabit in vivo: the peripheral blood (12% O2), the bone marrow (5% O2) and the TME (1% O2). NK cells incubated at 1% O2 have decreased proliferation and cytotoxicity compared to NK cells incubated at higher oxygen conditions. To assess what contributes to these changes, we conducted a mass cytometry (CyTOF) analysis, real time imaging assays, metabolic assays and gene expression (RNA-seq and ATAC-seq) analysis. We observed oxygen dependent changes in expression of activating receptors, Ki-67, perforin and granzyme. Hypoxia impacts aggregation of perforin and granzyme granules at the immune synapse. Under hypoxic conditions, NK cell metabolism resembles cancer cell metabolism with increased glycolysis, amino acid synthesis and central carbon metabolism. These changes are accompanied by mitochondrial defects. Gene expression analysis revealed that changes in NK cell metabolism and function are mediated at the epigenetic level by histone demethylases. Treatment with histone demethylase inhibitors rescued NK cell cytotoxicity as measured using an IncuCyte machine. These results indicate that NK cells who enter the TME are fundamentally different than those in the bone marrow or blood stream. The insights gained from this study will be leveraged to overcome hypoxia induced immune suppression in the TME to enhance NK cell-based immunotherapy for solid tumors.
Tipifarnib enhances eradication of acute myeloid leukemia by altering CXCL12/CXCR4 signaling in AML and by modifying the bone marrow microenvironment
Authors: Sterner Rosalie , Greif Lauren E , Khan Rehan , Kwok Tina , Peterson Kevin L , Kaufmann Scott H , Kessler Linda , Scholz Catherine , Gualberto Antonio , Hedin Karen E 
Published: February 2023

Abstract
The prognosis of acute myeloid leukemia (AML) remains poor in part due to the leukemic bone marrow microenvironment. Our lab has found that CXCL12, a chemokine abundant within the leukemic bone marrow microenvironment, induces apoptosis of AML cells expressing CXCR4, the receptor for CXCL12. However, this CXCL12/CXCR4-induced apoptosis is inhibited by differentiating osteoblasts, which protect AML cells from apoptosis in the bone marrow. Tipifarnib is a farnesyltransferase inhibitor shown to increase progression-free survival in AML patients that express high levels of CXCL12. Here, we report that tipifarnib inhibits the CXCL12/CXCR4-directed migration of AML cells via an ERK independent pathway. Furthermore, tipifarnib enhances CXCL12/CXCR4-mediated AML cell apoptosis via a mechanism that alters expression of apoptosis-regulating proteins. In addition, tipifarnib disrupts AML protection by osteoblasts, increasing AML cell apoptosis. Tipifarnib inhibits the osteoblast-mediated protection of AML cells via disrupting COL1A1 and TNAP, proteins essential for extracellular matrix production. In conclusion, tipifarnib alters the bone marrow microenvironment which is predicted to enhance eradication of AML via inhibiting CXCL12/CXCR4 directed cellular migration of AML cells, reducing the protective effects of differentiating osteoblasts by disrupting matrix protection proteins, and increasing CXCL12/CXCR4-mediated apoptosis.
Investigating the role of KLRG1 in regulatory T-cells and implications for anti-tumor immunity in Non-small Cell Lung Cancer
Authors: Bag Arup K , Schultz Andrew R , Goala Payal , Karreth Florian A , Adeegbe Dennis O 
Published: February 2023

Abstract
Although it is well established that CD4+CD25+Foxp3+ regulatory T cells (Tregs) dampen anti-tumor T cell responses in cancer, their molecular signature and clonal heterogeneity in NSCLC remains poorly understood. In this study, comparative phenotypic assessment of tumor and lymphoid tissue resident Tregs in KRAS/p53 mutant NSCLC mouse model revealed the presence of a distinct tumor infiltrating Treg population which express KLRG1. Compared to their counterparts, tumor associated KLRG1+ Treg showed higher expression of inhibitory receptors and activation markers, suggesting that KLRG1+ Tregs represent an activated subpopulation which may contribute to immunosuppression. Ex-vivo Treg suppression assay revealed that tumor associated KLGR1+ Tregs exhibited higher suppressive activity compared to KLRG1-Tregs. Although T cell development was normal in in-house generated KLRG1 KO mice including thymic and peripheral Treg numbers, KLRG1 KO Tregs showed less suppressive capacity compared to wild type, implying KLRG1 in Tregs functional program. Furthermore, α-KLRG1 antibody treatment in lung tumor bearing KP mice showed prolonged survival, accompanied by reduced proportion of KLRG1+ Tregs in the tumor. Finally, immunogenomic characterization of KLRG1 KO/wt and tumor infiltrating KLRG1+/− Tregs revealed distinct gene expression patterns and associated immune pathways. Collectively, our findings indicate that KLRG1 is not critical for Treg development and maintenance but may play a non-redundant role in Tregs differentiation and function in inflammatory settings. Therefore, targeting KLRG1 to curtail the inhibitory function of the highly suppressive KLRG1+ Treg subset could facilitate antitumor responses in NSCLC.
Perturbed function of natural killer cells by inflammatory cytokines in chronic myeloid leukemia
Authors: Kuznetsova Valeriya , Camacho Virginia , Patel Sweta , Matkins Victoria , Welner Robert S 
Published: February 2023

Abstract
Natural killer (NK) cells’ mature phenotype and counts positively correlate with prolonged treatment-free survival in chronic myeloid leukemia (CML), while dysfunctional NK cells are predictive of relapse in leukemic patients. However, the molecular drivers of NK cell alterations during leukemia remain unclear. Given an established role of inflammatory cytokines in CML progression, we define the leukemic environmental cues impacting NK maturation and function in a BCR-ABL chimeric mouse model of CML where NK cells do not carry the CML oncogene. Reflecting clinical observations, NK cell frequencies and numbers are severely decreased in leukemic mice. Moreover, terminally mature NK subsets are the most affected, implying preferential loss of cells with potent anti-leukemic activity. In addition, this phenotype suggests a shift toward the immature cytokine-producing state of NK development, which could favor the survival of leukemia-initiating cells. We map the transcriptional landscape of NK cells using single-cell RNA-sequencing from multiple tissues. NK cells exposed to CML microenvironment possess a lowered expression of surface activating receptors such as Nkp46 while upregulating inhibitory molecules Tim-3 and TIGIT. BCR-ABL reversion for six weeks partially restored these defects, consistent with findings in patients before and after TKI initiation. Finally, we found that leukemia-associated soluble factors dampen target-specific degranulation of NK cells by applying serum samples from healthy or leukemic mice. These data indicate that the altered cytokine milieu contributes to NK cell dysfunction and represents an optimal target for NK-boosting CML immunotherapies of leukemia.
NK-92 serial killers - characterization of potential limitations to cytotoxic capacity.
Authors: Navarrete-Galvan Lydia T , Guglielmo Michael , Sung Alexander , Smith-Gagen Julie , Lombardi Vincent , Merica Rebecca , Hudig Dorothy 
Published: February 2023

Abstract
The NK cell line NK-92 and transgenic variants are receiving attention as adoptive transfer immunotherapies to treat a range of malignancies. However, since NK-92s are themselves tumors, they require irradiation prior to transfer and are potentially susceptible to attack by patients’ immune systems. In the present study, we investigated NK-92 serial killing against various tumor cells as well as the effects of gamma-irradiation and ligation of the death receptor Fas (CD95) on NK-92 cells and the susceptibility of NK-92s to attack by primary NK cells. To evaluate serial killing, we used a 51Cr-release assay with very low effector to target (E:T) Raji, Daudi or K562 tumor cell ratios at 2, 4, 6, and 8 hour time points. NK-92s that were maintained in high concentrations of recombinant IL-2 were able to kill as many as 14 Raji cells per NK-92 cell in 8 hours at an E:T of 1:32. NK-92 cells retained high cytotoxic activity immediately after irradiation with 10 Gy but lost &gt;50% activity 24 hours after just 7 Gy irradiation. Despite strong expression of CD95, NK-92s maintained viability after 24 hours of Fas-ligation with an anti-CD95 IgM as compared to cells treated with an isotype control antibody. While NK-92s survived 16 hours of exposure to unstimulated primary NK cells, they were killed when exposed to IL-2 activated primary LAK cells. In conclusion, we recommend that adoptive transfer of irradiated NK-92 cells into patients be made as quickly as possible post-irradiation of the NK-92s. Genetic modification of NK-92 cell lines to secrete IL-2 as an autocrine growth factor may need reconsideration because in vivo activation of endogenous LAK cells by IL-2 could reduce the number of NK-92 cells within tumors and thus decrease therapeutic efficacy.
Expression of Tim-3 drives Treg to an effector-like state with enhanced suppressive activity and altered response to tumor
Authors: Banerjee Hridesh , Nieves-Rosado Hector , Murter Benjamin Matthew , Kane Lawrence P 
Published: February 2023

Abstract
Regulatory T cells (T reg) are mediators effective anti-tumor immunity. We and others previously reported that 40–60% percent of T reg-infiltrating head and neck cancer (HNC) and other tumors highly express Tim-3, compared with about 5% in lymphoid organs. Tumor-infiltrating Tim-3+ T reg also have enhanced suppressive function. Using a novel mouse model with cell type-specific inducible Tim-3 expression, we show here that expression of Tim-3 by T reg is sufficient to drive Treg to a more effector-like phenotype, resulting in enhanced suppressive activity and increased tumor growth. TIM-3 +T reg also seem to be metabolically altered driven by active signaling in ERK and m-TOR pathway. We further see that T reg specific deletion of TIM-3 leads to decrease in T reg frequency in tumor infiltrating lymphocytes and delay in tumor progression. These findings suggest to a possibility of using Tim-3 as a target to alter the tumor progression and clearance highliting a qualitatively different role for TIM-3 in T reg cells. Thus, we propose that Tim-3 regulates anti-tumor immunity at least in part through enhancement of T reg function. To our knowledge, this is the first example in which expression of a single co-stimulatory molecule is sufficient to drive differentiation of T reg and alter immune response in this manner.
Loss of Adar1 increases T cell migration and prolongs survival in ovarian cancer
Authors: Gomez Stephanie , Grundy Erin E , Kanholm Tomas , Hadley Melissa , McDonald James I , Rentia Uzma , Kobyra Julie , Palmer Erica , Saunthararajah Yogen , Chiappinelli Katherine B 
Published: February 2023

Abstract
Novel therapies are urgently needed for ovarian cancer (OC), the fifth deadliest cancer in women. OC is generally characterized by an immunosuppressive tumor microenvironment (TME) and less than 10% of patients respond to immune checkpoint blockade (ICB) therapy, though better OC prognosis is associated with high tumor-infiltrating CD8 T cells. DNA methyltransferase inhibitors (DNMTis) remove methylation and activate transcription of double-stranded (ds)RNA in OC. These dsRNAs are sensed in the cytoplasm and trigger the induction of type I IFN and also the transcription of interferon-stimulated genes (ISGs). Adenosine deaminase 1 (ADAR1) is an ISG that edits mammalian dsRNA with an A-to-I nucleotide change. These edited dsRNAs cannot be sensed by dsRNA sensors, and thus ADAR1 can inhibit this type I IFN response. Our preliminary RNA-seq analysis from human OC cell lines indicates that RNA editing by ADAR1 is increased after DNMTi treatment.
To test the hypothesis that Adar1 loss will amplify the DNMTi-induced IFN response in OC, we implanted mice with syngeneic ID8 murine OC Adar1 knockdown (KD) cells or control cells and treated with DNMTi. We show that DNMTi treatment reduces tumor burden and extends survival in this OC model. Loss of Adar1 significantly prolongs survival in this model, which is further improved with DNMTi treatment. DNMTi treatment of murine OC cells combined with Adar1 KD increases T cell migration in transwell migration assays and also enhances expression of chemokines CCL5, CCL2, and CXCL10. We are currently performing immunophenotyping analyses to interrogate the composition of immune cells in the TME in the mouse model of OC. These studies thus describe a role of Adar1 in the DNMTi-induced immune response in OC.