bluuebunny/crossref_metadata_2025_split huggingface dataset:
BAFF-R: A new target for depletion of pathogenic B cells in experimental lupus
Authors: Neeli Indira , Jha Kumar Abhiram , Moore Charles Edward , Marion Tony , Radic Marko 
Published: January 2023

Abstract
Lupus is an autoimmune disease in which autoantibodies with distinct specificities may arise. However, which autoantibodies contribute to specific manifestations in this multi-system disorder is unknown. We recently demonstrated that anti-CD19 CAR T cell therapy effectively depletes CD19+ B-cells and reduces anti-DNA antibodies in autoimmune mice, resulting in improved health and increased life span. Here, we analyzed autoantibody specificities in MRL/lpr mice both before and after 4 months of anti-CD19 CAR T cell treatment. Although antibodies to cardiolipin and histones decreased in parallel with anti-dsDNA in CAR T cell treated mice, autoantibodies to a ribonucleoprotein (RNP) autoantigen with a speckled nuclear distribution remained present. This result argues that anti-RNP IgG are unlikely to contribute to nephritis and skin inflammation, which markedly improved in the anti-CD19 CAR T cell treated mice.
The anti-CD19 CAR T cell treated mice were also used to identify potential B cell surface receptors that could serve as CAR T cell targets on kidney-infiltrating B cells. Kidney RNA transcripts, as measured by RT-PCR, showed only minor changes for CD11c, CXCR4, and BCMA between CAR T-treated and control mice, whereas the expression of BAFF-R decreased proportionally with CD19 in treated mice. Thus, the construction and testing of anti-BAFF-R CARs in mice may provide a useful step toward a potential new synergistic therapy for difficult to treat lupus in patients.
Research was supported by the Lupus Research Alliance and the UTHSC Maturation Award.
Chloroquine modulates inflammatory autoimmune responses through Nurr1 in autoimmune diseases
Authors: Park Tae-Yoon , Kim Kwang-Soo 
Published: January 2023

Abstract
For over a half-century the anti-malarial drug chloroquine (CQ) has been used as a therapeutic agent, alone or in combination, to treat autoimmune diseases. However, neither the underlying mechanism(s) of action nor their molecular target(s) are well defined. The orphan nuclear receptor Nurr1 (also known as NR4A2) is an essential transcription factor affecting the development and maintenance of midbrain dopaminergic neurons. In this study, using in vitro T cell differentiation models, we demonstrate that CQ activates TREG cell differentiation and induces Foxp3 gene expression in a Nurr1-dependent manner. Remarkably, CQ appears to induce Nurr1 function by two distinct mechanisms: firstly, by direct binding to Nurr1’s ligand-binding domain and promoting its transcriptional activity and secondly by upregulation of Nurr1 expression through the CREB signaling pathway. In contrast, CQ suppressed gene expression and differentiation of pathogenic TH17 cells. Importantly, using a valid animal model of inflammatory bowel disease (IBD), we demonstrated that CQ promotes Foxp3 expression and differentiation of TREG cells in a Nurr1-dependent manner, leading to significant improvement of IBD-related symptoms. Taken together, these data suggest that CQ ameliorates autoimmune diseases via regulating Nurr1 function/expression and that Nurr1 is a promising target for developing effective therapeutics of human inflammatory autoimmune diseases.
CRS free T cell therapy: T cell membrane anchored and tumor targeted IL12 (ATTIL12)-T cell therapy eliminates large tumors, overcomes heterogeneity, and avoids adverse effects
Authors: Hu Jiemiao , Zhang Wendong , Xia Xueqing , Yang Qing , Chen Yuhui , Fowlkes Natalie W , Livingston John Andrew , Zhang Zhongting , Mahadeo Kris M , Gill Jonathan B , Dotti Gianpietro , Kopetz Scott , Gorlick Richard , Li Shulin 
Published: January 2023

Abstract
Adoptive transfer of genetic engineered T cells becomes an effective approach in treating hematologic malignancies, but shows poor responses against solid tumors due to multifactorial reasons. Interleukin-12 exhibits robust T cell activating and anti-tumor efficacy. However, its clinical application was impeded due to the lethal adverse effects. We demonstrated previously that tumor targeted IL-12 (ttIL12) gene therapy avoids the toxicity of wildtype IL12 and effectively eliminate tumor metastasis. In this study, we developed a second generation of ttIL12— membrane-anchored ttIL-12 (attIL12)—to modify naive T cells, CD19CAR-T cells and OT1 T cells. This attIL12-modified T cells silent the toxicity of both IL12 and CAR-T cells associated cytokine release syndrome (CRS). Surprisingly, this ATTIL12-T cell therapy is able to eliminate multiple types of tumors with either homo- or hetero-generous environment or large size. We are in full speed to prepare a phase I clinical trial.
Engineering Chimeric Antigen Receptor T cells Against Immune Checkpoint Inhibitors PD-1/PD-L1 for Treating Pancreatic Cancer
Authors: Liu Chao Lien , Fan Ming Huei , Miao Carol H. , Liao Yi Jen 
Published: January 2023

Abstract
Pancreatic ductal adenocarcinoma (PDAC) is an aggressive disease with a 5-year survival rate of 6%. Major obstacles to successful treatment of pancreatic cancer are the immunosuppressive tumor microenvironment (TME) and antigenic complexity or heterogeneity. Programmed death-ligand 1 (PD-L1) is expressed on PDAC and immunosuppressed cells within the TME, providing suitable immunotherapy targets. We applied a chimeric antigen receptor (CAR) strategy to target immune checkpoint PD-1/PD-L1 interactions. Lentiviral vectors were used to express the extracellular domain of human PD-1 (PD1ACR) or the scFv region of aPD-L1 (PDL1CAR) that bind to PD-L1, and each was fused to intracellular signaling domains containing CD3 zeta, CD28, and 4-1BB (CD137). Both engineered CAR T cells recognized and eliminated PD-L1-overexpressing CFPAC1 cells efficiently at approximately 80% in vitro. Adoptive transfer of both CAR T cells enhanced T-cell persistence and induced specific regression of established CFPAC1 cancer &gt;80% in a xenograft model. Ki67 expression in tumors decreased, whereas proinflammatory cytokines/chemokines increased in CAR T-cell-treated mouse sera. PD1ACR and PDL1CAR obtained a similar therapeutic efficacy. Thus, these armed third-generation PD-L1-targeted CAR T cells confer antitumor activity and ability to combat T-cell exhaustion, providing a potentially new and innovative CAR T cell immunotherapy against pancreatic cancers.
Modular Antigen-Specific T Cell Suppression Assay for In Vitro Drug Screening
Authors: Kleiman Eden , Sierra Gloria , Daftarian Pirouz 
Published: January 2023

Abstract
T cell exhaustion (TEX) describes a T cell differentiation continuum of hierarchical progressive diminution of effector function. Many factors have been attributed to this including expression of multiple co-inhibitory receptors, altered transcription factor expression, epigenetic rewiring and dysregulated metabolism. Antigen persistence is necessary for driving TEX maintenance in both chronic viral infection and cancer settings. In addition to persistent antigen exposure, tumor-infiltrating lymphocytes (TILs) within the tumor microenvironment (TME) encounter immunosuppressive metabolic byproducts, suppressive cytokines, hypoxia and cellular debris which converge to suppress T cell function. These suppressed T cells are incapable of mounting an optimal anti-tumor response due to dampened proliferation, metabolic fitness and/or cytolytic capability. Our team has previously deployed usage of the recall antigen assay to screen immune checkpoint inhibitor (ICI) drug candidates. We have shown that in some donor PBMCs, ICI drugs such as pembrolizumab are able to boost antigen-specific T cell recall. We adapted this antigen recall assay to study TME-mediated T cell suppression. We show application of this TME-based recall assay by addition of adenosine which suppresses antigen-specific T cell expansion without altering T cell viability. Interestingly, T cell suppression was partially reversed with addition of CMV lysate. This assay is modular in that TME metabolites can be used alone or in combination, with the potential to screen for donors who are in vitro “responders” and “non-responders” to various drug classes.
The Force Awakens: Illuminating the Role of Kynurenine in Cancer Progression and Treatment
Authors: Ashoura Norah E , Dekker Joseph , Triplett Todd A , Garrison Kendra , Blazeck John , Karamitros Christos , Lamb Candice , Tanno Yuri , Ehrlich Lauren Ilyse Richie , Zhang Michelle , Manfredi Mark G. , Stone Everett , Georgiou George 
Published: January 2023

Abstract
Cancer is the second leading cause of death in the United States, with an estimated 40% of all Americans expected to be diagnosed with cancer in their lifetime. Despite progress in treatment options, major obstacles in current therapies must be overcome to limit their harmful side effects on patients. To evade immune clearance, many cancers elevate tryptophan (Trp) catabolism in the tumor microenvironment (TME) by upregulating the enzyme indoleamine 2,3-dioxygenase (IDO). As a result, cancer cells (1) monopolize extracellular tryptophan and (2) release L-kynurenine into the TME, an independent immune signal. This change in metabolism generates immune suppression in the TME, but whether the cause arises from Trp depletion or the accumulation of the IDO product kynurenine (L-Kyn) remains highly controversial. Kynurenine is known to induce immunosuppressive phenotypes through aryl hydrocarbon receptor (AhR) activation. However, the functional role of this binding is still poorly understood, despite an explosion of interest and information in the field. Our strategy uses the enzyme kynureninase (KynU) to degrade kynurenine into non-toxic products, thereby restoring anti-tumor immunity without harming the tryptophan metabolism of healthy cells. Moreover, this work aims to determine kynurenine’s molecular effect on T cells to better understand why its depletion successfully relieves tumor burden.
Antifungals improve anti-PD1 efficacy in a murine breast cancer model
Authors: Kershaw Kathleen , Henson Regina M. , Abrajan Brandon , Choi Rachel Y. , Underhill David M. , Shiao Stephen L. 
Published: January 2023

Abstract
Every year over 200,000 North American women are diagnosed with breast cancer, which is the second leading cause of cancer death among women. The introduction of effective immunotherapies has revolutionized patient care in many cancers, including breast, but only a subset of patients exhibit clinical response. Factors determining a patient’s response to immunotherapy is an area of intense research. The microbiome is one potential contributing factor. The human intestinal microbiome is comprised largely of bacteria, but also harbors fungi, viruses and archaea that undoubtedly have significant biological functions. Recent reports have detailed the association of specific strains of bacteria with positive response to PD-1/PDL1 therapy in several cancers however, little is known regarding the involvement of commensal fungi. Previous work in our lab found a high diversity of fungi associated with the human gut, which interact with the immune system and influence the severity of gastrointestinal inflammation and allergic airway disease. Using a syngeneic murine model of triple negative breast cancer, we found that antifungal treatment sensitizes previously resistant tumors to anti-PD1 therapy, leading to a significant decrease in growth rate as well as increased survival compared to anti-PD-1 treatment alone. Antifungal treatment altered the microbiome composition as evidenced by 16s and ITS1 sequencing of fecal pellets, as well as altered the tumor immune compartment to promote anti-tumor activity when combined with anti-PD1. These data suggest fungi have a role in shaping the immune-tumor microenvironment and provide insight into how perturbation of the microbiome might improve response to immunotherapy.
Determining the mechanism of the immune response to histotripsy ablation of pancreatic cancer
Authors: Hendricks Alissa Danielle , Brock Rebecca Michaela , Gannon Jessica , Zeher Allison , Sereno Jacqueline , Vlaisavljevich Eli , Allen Irving C 
Published: January 2023

Abstract
Histotripsy is a novel, non-thermal, image guided ablation modality that can rapidly kill cells in a targeted region with millimeter precision. Previous work established that there is an immune response to histotripsy ablation, however a mechanism behind this response has not been established. For the current study, we utilized an in vitro model, Pan02 (pancreatic adenocarcinoma), transfected with a known antigen (hemaglutanin/HA), and verified findings in vivo. Cells were ablated at therapeutic and sub-therapeutic doses to determine the release of damage associated molecular pattern (DAMP). For both treatments, gel electrophoresis confirmed the release of damaged DNA, and western blots verified the release of intact HMGB1 and HA proteins. To determine the efficacy of the released DNA to stimulate an immune response, we exposed naïve macrophages to isolated DNA from the treatment supernatant and showed activation by 24 hours. Additionally, to show the potential for a systemic immune response, complete supernatant was given to naïve dendritic cells, that were in turn co-cultured with naïve T cells. CD4+ and CD8+ T cell activation was confirmed with IL-2 and INFg ELISAs as well as flow cytometry. To verify these findings, mice with Pan02 tumors were treated, and demonstrated similar DAMP and antigen release. Further, changes in immune cell populations within the tumor were consistent, with increased macrophages and dendritic cells 24 hours and 7 days and in T cell populations at 7 and 14 days after treatment. These results show the role of specific DAMPs and immune cell populations in responding to histotripsy ablation. Overall, these results indicate a robust immunomodulatory response to histotripsy tumor ablation.
NKG2A inhibits the anti-tumor CD8 T cell immune response elicited by radiotherapy
Authors: Battaglia Nicholas Griffin , Caldon Johnathan J. , Okoshi Ethan N. , Gerber Scott A. , Lord Edith M. 
Published: January 2023

Abstract
More than half of patients with solid tumor malignancies undergo treatment with radiotherapy (RT). In addition to causing direct tumor cell death, RT results in release of tumor antigen and damage associated molecular patterns that elicit a CD8 T cell and IFN-g dependent anti-tumor immune response. To further bolster this response, RT has been combined with immune checkpoint inhibitors such as anti-PD-1 and anti-CTLA-4. Despite a modest increase in survival when these therapies are used alone or together in conjunction with RT, many patients fail to respond to treatment, suggesting other mechanisms of immune suppression exist in the tumor microenvironment (TME). Using syngeneic B16 F10 melanoma and C38 colorectal adenocarcinoma models, we have studied the role of the inhibitory receptor NKG2A. We have observed that NKG2A is expressed only on tumor infiltrating lymphocytes (TILs) and expression of this receptor does not change with RT. Furthermore, RT increases intratumoral expression of the ligand for NKG2A, Qa-1b, as does IFN-g stimulation in vitro. Blockade of NKG2A alone through use of B16 F10 cells lacking Qa-1b expression or blocking antibodies did not significantly increase survival of mice treated with RT. Further analysis revealed that among CD8 TILs, only a minority of cells express NKG2A alone, with most TILs co-expressing NKG2A and PD-1 or PD-1 alone. Thus, we combined anti-NKG2A and anti-PD-1 blockade and observed increased survival in mice treated with RT, whereas either therapy alone was ineffective. These results suggest that NKG2A blockade could be combined with RT and existing immunotherapies clinically to improve patient response. Supported by awards T32AI007285 from the NIAID and R01CA028332 from the NCI.
Tumor cell-intrinsic PD-L1 signals promote DNA damage responses that mediate resistance to Chk1 and PARP inhibitors in vivo
Authors: Kornepati Anand V , Zhang Deyi , Padron Alvaro S , Boyd Jacob T , Deng Yilun , Osta Erica G , Reyes Ryan M , Shen Hailian , Wang Juan , Kari Suresh , Clark Curtis A , Hu Yanfen , Li Rong , Gupta Harshita B , Zhao Weixing , Curiel Tyler J 
Published: January 2023

Abstract
Tumor PD-L1 mediates diverse cell-intrinsic signals that increase cancer pathogenesis including the DNA damage response (DDR). Although immune consequences of tumor-extrinsic PD-L1 blockade are known, novel therapeutic vulnerabilities from inhibiting tumor-intrinsic PD-L1 signals are little studied. To assess tumor-intrinsic PD-L1 DDR control, we depleted PD-L1 by CRISPR/Cas9 or shRNA (PD-L1KO) in mouse B16 (melanoma), 4T1 (breast), ID8agg (ovarian), and human bladder RT4 cancer lines. We identified specific DDR signaling defects in PD-L1KO versus control (con) cells after exposure to distinct DNA damaging agents. 24h gemcitabine (or 2 Gy X-ray) in vitro in PD-L1KO vs. con RT4 cells elevated γH2AX (DNA damage marker) and reduced Chk2 DDR gene expression but not ATM, ATR, Chk1, or BRCA1 (immunoblots), and decreased BRCA1/p-RPA32 nuclear foci formation (confocal) altogether suggesting tumor-intrinsic PD-L1 promotes specific DDR/DNA repair pathways. PD-L1 loss could thus predict synthetic lethality to DDR inhibitors (DDRi) Chk1i or PARPi. PD-L1KORT4, ID8, and 4T1 cells were more sensitive to small molecule Chk1i in vitro (&gt;15 fold vs. con, MTT) and in vivo in NSG mice. Chk1i and PARPi led to γH2AX increase in PD-L1KO B16, RT4, and 4T1 vs. sham treated tumor cells as predicted by DDR defects. Genetic cellular PD-L1 depletion, but not anti-PD-L1 (extrinsic), strongly sensitized B16 and 4T1 tumors to PARPi in vivo. Strikingly, PARPi had no effect on PD-L1KO B16 in RAG2KO mice despite treating WT mice, indicating a strong immune component to DDRi treatment efficacy. Thus, DDRi plus inhibiting tumor intrinsic PD-L1 signals could improve immunotherapy in immunotherapy-sensitive or resistant tumors.
Targeting GARP-TGFβ axis enhances anti-tumor efficacy of immune checkpoint blockade therapy
Authors: Li Anqi , Wu Bill X. , Riesenberg Brian P. , Li Zihai 
Published: January 2023

Abstract
Although immune checkpoint blockade (ICB) has emerged as a promising cancer therapy, a majority of cancers, such as pancreatic cancer and triple negative breast cancer, fail to respond to ICB. A potential mechanism is the accumulation of transforming growth factor-β1 (TGFβ1) in tumor microenvironment (TME) which drives immune dysfunction by inducing regulatory T cells (Treg), excluding and inhibiting the function of effector CD8+ T cells. Glycoprotein-A repetitions predominant protein (GARP) is a cell surface docking and activating receptor for latent TGFβ1 and highly expressed in breast cancer cells, Tregs and platelets. Previous work from our lab has demonstrated a critical role for GARP in dampening anti-tumor immunity via TGFβ1 pathway. To validate if GARP is a therapeutic target, we generated an anti-human GARP antibody (4D3). We found that 4D3 blocked GARP-latent TGFβ1 interaction and limited active TGFβ1 release. We expressed human GARP in murine 4T1 breast cancer cells. In this orthotopic syngeneic mouse tumor model, combination therapy with 4D3 and anti-PD1 antibody suppressed tumor growth and metastasis more effectively than treatment with either agent alone, resulting in significantly enhanced survival. Mechanistically, the combination therapy reduced p-SMAD3 expression as well as collagen deposition in the TME. In addition, 4D3 treatment reduced tumor-associated macrophages and their PD-L1 expression. Collectively, our preclinical data demonstrated that simultaneous blockade of the PD1 and GARP-TGFβ1 axis elicits potent and superior anti-tumor efficacy relative to monotherapies, suggesting a potentially more effective strategy against cancers that are resistant to current immune checkpoint blockade.
Characterization of the immune microenvironment in melanoma patients resistant to targeted and immunotherapies by a multidisciplinary approach
Authors: Veigas Florencia , Mahmoud Yamil D , Flores Ana , Morales Rosa , Gatto Sabrina , Saez Juan Manuel Perez , Fernandez Elmer , Rabinovich Gabriel A , Girotti María Romina 
Published: January 2023

Abstract
Most melanoma patients who develop resistance to targeted therapies do not respond to subsequent immunotherapy. Therefore, establishing which patients would derive clinical benefit from immunotherapy is a compelling clinical question. We performed a meta-analysis of published transcriptome datasets from melanoma biopsies before and after development of resistance to BRAFi or BRAFi/MEKi. We observed an increase in M2-macrophages infiltrate by CIBERSORT in relapsed biopsies as well as an increase in T-cell exhaustion markers and immunosuppressive cytokines. Our preliminary findings show changes in the galectin/glycan axis in melanoma resistance to BRAFi/MEKi. Particularly, we observed that the expression of Galectin-1 (Gal-1) is increased in human melanoma resistant cells. We analyzed single-cell and bulk RNA-Seq gene expression profiles from melanoma biopsies before and during treatment with anti-PD-1/anti-CTLA-4 and found that macrophages and CD8 T-cells of non-responders upregulate a specific glycosylation-related signature. Macrophages from non-responding patients show an immunosuppressive expression profile accompanied by the appearance of a novel sub-group during treatment associated with resistance. We hypothesize that there are Gal-1/glycan interactions that promote an M2 immunosuppressive microenvironment in resistant tumors preventing response to immunotherapy. Finally, we developed MIXTURE, a novel algorithm to infer the tumor immune infiltrate that overcomes CIBERSORT limitations. MIXTURE analysis revealed a differential immunosuppressive infiltrate in melanoma biopsies with high intratumoral heterogeneity and in biopsies from non-responding patients to anti PD-1 immunotherapy.
Molecular iodine activates cytotoxic immune response in breast cancer tumor microenvironment
Authors: CUENCA OLGA , GONZÁLEZ EVANGELINA DELGADO , VELASCO CARMEN ACEVES 
Published: January 2023

Abstract
Present work analyzes the tumoral immune response of oral molecular iodine (I2) supplementation with the conventional treatment (5-fluorouracil/epirubicin/ cyclophosphamide or taxotere/epirubicin; FEC/TE) in patients with breast cancer from Hospital General of Queretaro, Mexico. Patients diagnosed with stage II were given placebo (vegetable colored water) or I2 from diagnosis to mastectomy (7 to 40 days) while patients with stage III received placebo or I2 with neoadjuvant FEC/TE treatment (40–70 days). Transcriptomic analysis and GESA studies showed that I2 supplementation alone or in presence of FEC/TE up-regulate Th1 and Th17 differentiation pathways, activation by T-cell receptor, cytotoxicity by NK and CD8+, lymphocyte migration and formation of tertiary lymphoid structures. Deconvolution analysis indicate an increase in macrophages/dendritic cells in I2 group while in FEC/TE+I2 group exhibited increased B cells fractions. Analysis of Th1/Th2 response markers showed significant increase expression in Tbx-21 (RT-PCR) and Interferon gamma (immunohistochemistry) in the FEC/TE+I2 group. This anti-tumor immune activation correlated with the longer disease-free period (43%) and 5 years-survival (60%) observed in FEC/TE+I2 patients. Studies to analyze the effect of I2 supplementation in immunotherapies are in course.