bluuebunny/crossref_metadata_2025_split huggingface dataset:
Therapeutic regulation of autoreactive B cells using bispecific molecules
Authors: Griffith Amber Simone , Apley Kyle , Downes Grant , Johnson Stephanie , Ross Patrick , DeKosky Brandon , Farrell Mark , Berkland Cory , Kendall Peggy L 
Published: January 2023

Abstract
Type 1 diabetes (T1D) occurs when autoreactive T cells destroy insulin-producing beta cells. Anti-insulin B cells promote this disease, acting as essential antigen-presenting cells. Rituximab, which globally depletes B cells, provided temporary preservation of insulin production in early onset T1D, but increases susceptibility to infection. Antigen-specific immune therapy (ASIT) may be a viable option for treating T1D by targeting anti-insulin B cells for removal from the B cell repertoire. In this study we investigate the efficacy of soluble antigen arrays containing insulin (SAgAins) developed by the Berkland lab. These constructs consist of a polyethylene glycol (PEG) backbone with four linker arms for bioconjugation. We conducted in vitro experiments to investigate the effects of constructs containing insulin and a trisaccharide ligand (CD22L) for inhibitory B cell receptor CD22 on transgenic anti-insulin B cells. We found that incubating insulin-PEG with anti-insulin B cells stimulates proliferation and CD86 upregulation. However, when CD22L is added (CD22L-PEG-insulin) the expression of CD86 is dampened. This suggests that this novel construct harnesses the regulatory power of CD22 to dampen activation of B cells in an autoantigen-specific manner.
Impact of Swine Leukocyte Antigen on renal endothelial cells in pig-to-non-human primate xenotransplantation in vitro
Authors: Wang Zheng-Yu , Reyes Luz , Estrada Jose , Tector Mathew , Tector Alfred 
Published: January 2023

Abstract

BACKBROUND
The development of a pig lacking the GGTA1, B4GNT2 and CMAH genes (TKO) that produce the three known glycan xenoantigens has provided the decreased background human antibody binding to TKO cells. It can be easily detected for antibody binding to sufficiently low levels of any additional xenoantigens expressed on the pig cells, including swine leukocyte antigen (SLA). The object of this study is to evaluate the role of SLA class I and II in the pig-to-non-human primate (NHP) xenotransplantation in vitro at an organ-specific cell level using renal endothelial cells (REC).

METHODS
Characteristics of genetically-edited REC and Interferon gamma (IFN-γ)-activated REC were analyzed using flow cytometry. Rhesus IgG/IgM binding to REC with or without SLA class I, or SLA class II expression were examined using flow cytometric crossmatch and complement-dependent cytotoxicity assay (CDC).

RESULTS
SLA I-deficient REC with GGTA1/B4GNT2/CMAH TKO background had significantly lower Levels of antibody binding (p=0.02 and p=0.0001, respectively for IgG and IgM) using 25% rhesus sera (n=14). SLA class II expression on SLA I-deficient GGTA1/B4GNT2 REC was induced following 3-day treatment of IFN-γ. These cells with SLA class II expression had significantly increased IgM binding by crossmatch (p=0.002), and had significantly high levels of cell lysis by CDC assay (p&lt;0.0001) using 25% rhesus sera (n=11).

CONCLUSIONS
Gene editing of the SLA class I and II on pig organs might protect against chronic antibody-mediated rejection via reducing antibody binding to REC in pig-to-NHP renal xenotransplantation. Porcine REC cell lines will be a useful reagent for the further study of xenoimmunology.
Supported by Partial 5R01AI126322
Targeting temporal metabolic needs of T follicular helper cells to treat cGVHD utilizing a mitochondrial pyruvate carrier inhibitor
Authors: Mohamed Fathima A , Rhee Stephanie Y , Ly Joanna , Aguilar Ethan G , Sage Peter T , Schumacher Tanner , Thangavelu Govindarajan , Zaiken Michael C , Liu Juan , Mereddy Venkatram , Locasale Jason W , Blazar Bruce R 
Published: January 2023

Abstract
The curative potential of allogeneic hematopoietic stem cell transplantation is limited by development of chronic graft versus host disease (cGVHD) and metabolic regulation of immune cells is a promising therapeutic target. During cGVHD, interactions between T follicular helper (TFH) and germinal center B (GCB) cells result in production of alloreactive class-switched antibodies that attack host tissues, causing multi-organ damage and tissue fibrosis. We show the metabolic dependencies of TFH change throughout cGVHD progression and targeting mitochondrial pyruvate transport is an effective strategy to mitigate disease. Pyruvate, the end product of glycolysis, is transported by the mitochondrial pyruvate carrier (MPC) for use in the tricarboxylic acid cycle and oxidative phosphorylation (OXPHOS). TFH require glycolysis in order to induce a GC reaction, both in an immunization model and in an in vivo systemic cGVHD model with bronchiolitis obliterans (BO). Furthermore, Seahorse data from TFH extracted early in disease development in the BO model demonstrate increased dependency on glycolysis. But Seahorse data also indicates TFH undergo a shift away from glycolysis and toward OXPHOS as cGVHD continues. Based on the central role of MPC in tying pyruvate to multiple metabolic pathways, we examined an inhibitor of MPC (7ACC1) in reducing TFH activity. Administration of 7ACC1 resulted in inhibition of GCB production of class switched antibody in an in vitro assay. These findings were additionally supported in vivo, whereby daily treatment after establishment of cGVHD resulted in a decrease in disease severity. These data provide evidence that inhibiting mitochondrial pyruvate transport is a viable therapeutic strategy for cGVHD.
This work was supported by grants from National Institutes of Health, National Institute of Allergy, and Infectious Diseases R37 AI34495, T32 AI007313; National Heart, Lung, and Blood Institute R01 HL56067, R01 HL11879, R01 HL115114; National Cancer Institute P01 CA065493
A comparison of monoclonal antibodies against murine PD-1 and PD-L1
Authors: Bu Melissa Tian , Yuan Long , Klee Alyssa , Freeman Gordon J 
Published: January 2023

Abstract
Blockade of the PD-L1/PD-1 pathway has proven to be a broadly effective cancer immunotherapy. FDA-approved therapeutic monoclonal antibodies (mAb) targeting the pathway have high affinity, blocking capacity, and low antibody effector activity. A number of rat anti-mouse mAbs have been used to model cancer immunotherapy in mouse models. We set forth the amino acid sequences of mAbs specific for mouse PD-1 (29F.1A12) and PD-L1 (10F.9G2) and compare their avidities, blocking capacities, biological activities, and epitope recognition with other commonly used mAbs. Of note, we found that 29F.1A12 had an approximately 100-fold better affinity and blocking capacity than RMP1-14. Further manipulation of these sequences should facilitate better modeling of immunotherapy in mouse models and the generation of novel agents.
Supported by grants from the National Institutes of Health (P01 AI56299 and P01 CA236749).
MTAP deficiency reshapes the tumor immune landscape in lung cancer
Authors: Li Ji-Min , Zhang Jun , Chang Wen-Hsin , Hsu Ssu-Wei , Chen Ching-Hsien 
Published: January 2023

Abstract
Methylthioadenosine phosphorylase (MTAP) deficiency occurs in a broad range of malignancies; notably, loss of this metabolizing enzyme was reported to be associated with poor survival in cancer patients. However, the mechanisms underlying tumor progression due to MTAP loss are yet to be elucidated. Utilizing RNA-sequencing, reverse phase protein arrays, and Luminex assays, we demonstrated that MTAP deficiency alters tumor-intrinsic, immune-related pathways and reprograms cytokines toward a tumor-favorable environment in both lung and kidney cancer. In addition, MTAP-knockout cells exhibited a marked increase in the immune checkpoint protein PD-L1. Upon co-culturing primary T cells with cancer cells, we found that MTAP loss-mediated PD-L1 upregulation inhibited T cell-mediated killing activity and induced several T cell exhaustion markers. In two xenograft tumor models, we showed that the average volume of tumors derived from MTAP-deficient cells is slightly greater than that of MTAP-proficient tumors. Surprisingly, a remarkable increase in tumor size was also observed in humanized mice bearing MTAP-deficient tumors, as compared to their MTAP-expressing counterparts. Following immunophenotypic characterization of tumor-infiltrating leukocytes by mass cytometry analysis, compared to MTAP-expressing tumor xenografts, MTAP-deficient tumors was found to display decreased immune infiltrates that consist of abundant immunosuppressive cells and lower portions of both T lymphocytes and natural killer cells. Taken together, our results suggest that MTAP deficiency reconstructs the tumor immune microenvironment via cytokine reprogramming and immune remodeling, promoting tumor progression and immune evasion.
This work was supported by grants funded by the Department of Defense Kidney Cancer Research Program (KCRP) W81XWH1910831 (log# KC180170) and the California UCOP Tobacco-Related Disease Research Program (TRDRP) T29IR0704.
Hematopoietic protein RELT is upregulated in human breast and lung cancers and binds the cytoskeletal protein Filamin A.
Authors: Cusick John K , Yanumula Anusri , Shyu Yennie , Wang Wenjia , Abram Alyssa , Alcaide Jessa , Zhou James , Gillespie Brittany , Senderovich Michelle , Budhiraja Anuj , Qureshi Vaniya , Bilasy Shymaa , Shi Yihui 
Published: January 2023

Abstract
Receptor Expressed in Lymphoid Tissues (RELT), is a human TNFR superfamily member (TNFRSF19L) expressed most prominently in the hematopoietic system. Previous reports using RELT knockout mice indicate that RELT functions in part by inhibiting the proliferation and activation of T lymphocytes. RELT is an orphan receptor that has two homologous binding partners, RELL1 and RELL2, and collectively, these three proteins are referred to as RELT family members. This study sought to identify novel protein interactions with RELT family members and to compare the expression of RELT in normal and diseased human tissues. A yeast two-hybrid screen utilizing RELL1 as bait identified the cytoskeletal protein Filamin A (FLNA) as a prospective binding partner and physical interaction between FLNA and RELT family members was confirmed by in vitro co-immunoprecipitation. A truncated mutant of FLNA disrupts the ability of RELT family members to activate the p38 pathway and blocks the ability of RELT to induce death in human epithelial cells. Western blotting revealed endogenous RELT family members are most significantly upregulated in the breast cancer cell lines MDA-MB-231 and MCF-7, as well as the lung cancer cell line H358. Preliminary immunohistochemistry results reveal a higher intensity of RELT staining in both breast cancer and lung cancer human biopsies versus non-malignant tissue. Additionally, overexpression of RELT family members enhanced apoptosis in MDA-MB-231 and H358 cells as measured by X-gal morphology and luciferase assays. Collectively, these results further our understanding of signal transduction pathways associated with RELT family members and report the novel finding that RELT is upregulated in human breast and lung cancers.
Supported by California Northstate University, College of Medicine, Seed Grant.
An Emerging and Previously Unrecognized Paradigm of Cxcl12/Cxcr4 Pathway Implication in Breast Cancer Metastasis
Authors: Lagou Maria K , Sanchez Luis R , Duran Camille L , Burt Joseph , Chen Xiaoming , Lin Yu , Eddy Robert , Harney Allison S , Entenberg David , Condeelis John S , Oktay Maja H , Karagiannis George S 
Published: January 2023

Abstract
The Cxcl12/Cxcr4 signaling axis promotes metastasis in mouse models of breast carcinoma and is linked with increased metastatic risk in humans. Prior studies have linked Cxcl12/Cxcr4 to breast cancer cell homing and survival at metastatic sites. However, the precise mechanism via which Cxcr4+ breast cancer cells escape primary tumors, which also express Cxcl12, is poorly understood. By using a novel imaging method for visualizing chemokine gradients, we show that Cxcl12 is concentrically expressed around cancer cell intravasation doorways, known as Tumor Microenvironment of Metastasis (TMEM). Using distance analysis algorithms, we also show that TMEM-mediated Cxcl12 gradients contextually associate with Cxcr4+ breast cancer cells migrating towards TMEM doorways. As such, pharmacological inhibition of the Cxcl12/Cxcr4 axis significantly abrogates translocation of Cxcr4+ cancer cells to TMEM doorways, thus suppressing metastatic dissemination. However, the targeted elimination of Cxcr4 from breast cancer cells paradoxically results in a suboptimal response compared to pharmacologic inhibition, implying the existence of a compensatory mechanism. Using clodronate-mediated macrophage depletion, we demonstrate that, in the absence of Cxcr4 expression in tumor cells, accompanying Cxcr4+ macrophages may still “read” TMEM-generated Cxcl12 chemotactic gradients. Indeed, Pdgfrb+ stromal cells may express low levels of Cxcl12, but TMEM-generated Cxcl12 gradients are primarily linked to a subset of Iba1+ perivascular tumor-associated macrophages. Overall, our data reveal a new paradigm for the implication of Cxcl12/Cxcr4 in early stages of metastasis and propose an avenue for rationalized antimetastatic treatments.
Supported by Recruitment Funds RECR 3A3218
Multi-modal immune profiling of mucinous ovarian carcinoma: analysis from the Ovarian Tumor Tissue Analysis/Multidisciplinary Ovarian Cancer Outcomes Group consortia
Authors:   Ovarian Tumor Tissue Analysis Consortium, Meagher Nicola S , Hamilton Phineas T , Anglesio Michael S , Harris Holly R , Konecny Gottfried E , Schildkraut Joellen M , Talhouk Aline ,   Australian Pancreatic Genome Initiative, Friedlander Michael L , Pearce Celeste L , Pike Malcolm C , Köbel Martin , Doherty Jennifer A , Goode Ellen L , Nelson Brad H , DeFazio Anna , Ramus Susan J 
Published: January 2023

Abstract
Mucinous ovarian carcinoma (MOC) is a rare histotype of ovarian cancer. Women with advanced disease (FIGO stage III/IV) have very poor outcomes, with 5-year survival of &lt;20%. Little is known about the anti-tumor immune response in MOC, limiting potential immunotherapeutic options for patients.
Immune data from 3 platforms were analysed. A NanoString mRNA expression panel with 39 immune-related genes (n=60 MOC, n=4198 other ovarian histotypes), a NanoString mRNA plexset with PD-1, PD-L1 (n=241 MOC, n=115 upper and lower gastrointestinal (GI) cancers), and 2 multicolor immunofluorescence (mcIF) panels (n=121 MOC) for CD68, PD-L1, PD-1, CD8 and FOXP3. Data overlapped all platforms for 22 MOC. Immune marker expression by ovarian cancer histotype was measured. Uni- and multivariable Cox Proportional Hazards assessed overall survival (OS) within MOC. Hierarchical clustering looked for subgroups of patients based on co-expression of immune genes.
Compared to more common high-grade serous ovarian cancer (HGSC) and GI cancers, MOC were less immunologically active. High IGHM expression was associated with poorer OS (HR 1.31 (95% CI 1.02–1.69), p=0.033). High tumor densities of FOXP3 and PD-1 positive cells conferred poorer OS in a model with age, stage and site; however, these and PD-L1 and CD68 were associated with grade. The 39 gene NanoString and mcIF datasets revealed 4 clusters, roughly translating to immune cold, moderate and hot (groups 3,4). mcIF analysis revealed a subset with PD-L1 expression in the absence of T-regulatory cell infiltrates.
This study provides novel insights into the MOC immune landscape. Despite relatively low immune activity overall, immunotherapy could be explored for a subset of patients with MOC.
Supported by NSW Ministry of Health and UNSW Sydney, through the NSW Health PhD Scholarship Program 2017-2022; Translational Cancer Research Network top-up scholarship 2021
Upregulated Ly-6 gene expression is associated with poor overall survival in uterine corpus endometrial carcinoma patients
Authors: Rathbun Luke , Magliocco Anthony , Bamezai Anil K 
Published: January 2023

Abstract
The human Ly-6 (hLy-6) gene family has recently gained interest for its possible role in tumor progression. High expression of hLy-6 genes is associated with poor overall survival in pancreatic ductal adenocarcinoma patients. To expand upon this work, we have carried out in silico analyses of all known hLy-6 gene expression in other cancers using TNMplot which shows differential gene expression in cancer tissues. We have also analyzed the patient survival by Kaplan-Meier plotter after mining the TCGA database. We report that upregulated expression of several hLy-6 genes is associated with poor cancer patient survival in many other cancer types, especially uterine corpus endometrial carcinoma (UCEC). Importantly, the expression of a number of hLy-6 genes is elevated in UCEC tissue when compared to the expression in normal uterine tissue. Further analysis of tumor-specific expression of the hLy-6 gene family is needed to uncover the function of Ly-6 proteins as well as the signaling pathways these proteins trigger that endow tumor survival and poor patient prognosis. Human Ly-6 gene products can potentially be used as biomarkers for cancer detection and as tumor associated antigens to identify high-risk cancer patients and appropriately design and/or calibrate cancer treatments for such patients.
Analysis of chemokine network in primary human glioblastoma
Authors: Lachota Mieszko , Zielniok Katarzyna , Gozdz Agata , Szpak Patrycja , Kalaszczynska Ilona , Zagozdzon Radoslaw 
Published: January 2023

Abstract
Chemokines play an integral role in cancer immunoevasion by modulating leukocyte infiltration. They selectively increase the migration of immunosuppressive cells while impairing the infiltration of cytotoxic lymphocytes with potent antitumor activity. Some cancer-associated chemokines may also enhance tumor progression by acting as growth factors or promoting epithelial-mesenchymal transition.
Here, we acquired fresh human glioblastoma (GBM) tissues to define chemokine expression patterns in GBM tumors by integrating bulk and single-cell RNA sequencing of both tumor and healthy brain tissues along with Luminex secretome profiling of GBM primary cell cultures. Extensive mapping of GBM-associated chemokines has enabled us to gain insight into intratumoral chemokine-mediated cell-to-cell communication. Among others, we identified CCL7 and midkine (MDK) to play a crucial role in the formation of the hostile tumor microenvironment (TME) in GBM. We hypothesize that these chemoattractants can facilitate the GBM tumor growth by recruiting M2 macrophages, promoting epithelial-mesenchymal transition, and activating the prosurvival PI3K-Akt pathway. Indeed, high expression of both chemokines was linked to poor overall survival.
Additionally, by integrating the data, we further identified the cellular source of the most abundantly secreted chemokines in GBM tumors. Since immune cells require TME infiltration to mediate an appropriate antitumor response, impaired tumor infiltration is one of the critical limiting factors for cancer cell therapy. Our chemokine atlas provides novel targets to enhance adoptive cell therapies by improving the potential of effector cells for tumor homing and anticancer activities in GBM.
This research was funded by the National Science Centre, Poland (2020/37/B/NZ6/02191), the Polish Ministry of Science and Higher Education (grant number DI2018 020548), and the National Centre for Research and Development (STRATEGMED3/307326/6/NCBR/2017).
Leucine-rich repeats and immunoglobulin-like domains protein 1 is highly expressed on tumor-infiltrating T cells and may play a role in modulating T cell activation
Authors: Ta Hieu M , Roy Dia , Zhang Keman , Dong Juan , Gilmour Cassandra , Yoo Seong-Keun , Alban Tyler , Makarov Vladimir , Parthasarathy Prerana Bangalore , Chan Timothy , Wang Li 
Published: January 2023

Abstract
Immune checkpoint receptors (ICRs) regulate T cell-mediated immune responses to self - and neo-antigens, thereby play an important role in the development of autoimmunity and immune responses to infections. In cancer, ICRs such as PD-1 and CTLA4 impair immuno-surveillance against neoplastic cells, while other costimulatory molecules like ICOS, OX40 could augment the immune responses. Immunotherapy that targets either inhibitory or costimulatory immune checkpoint have become the breakthrough therapy for cancer treatment. However, the efficacy of this class of therapy is limited to a fraction of patients, pointing the need to overcome non-redundant immuno-inhibitory pathways or discover new immuno-costimulatory pathways. This study identified the expression of Leucine-rich repeats and immunoglobulin-like domains protein 1 (LRIG1) in activated CD4+ and CD8+ T cells. LRIG1 is a type-I transmembrane protein with 1092 amino acids (120 kD) and ubiquitous tissue expression patterns. We show that LRIG1 expression is not detectable on naïve CD4+ and CD8+ murine T cells but is induced upon T cell receptor (TCR) stimulation in vitro, and in tumor infiltrating lymphocytes (TILs) from multiple murine models. We also detected LRIG1 expression on CD4+ and CD8+ TILs in several types of cancer patients such as non-small-cell lung cancer, breast cancer, melanoma, B cell lymphoma. LRIG1 was originally identified as a tumor suppressor gene in a variety of tissues, but its expression and function have not been explored in immune cells. Our data suggest that LRIG1 is highly expressed in TILs in multiple cancer types and might play a role in regulating anti-tumor immune response
This project is also supported by fundings 2R01CA164225 (NCI), 1R01CA223804 (NCI), and American Cancer Society (ACS) RSG-18-045-01
CD4+ T cells mediate non-canonical rejection of major histocompatibility class-I deficient pancreatic tumors independently of CD8+ T cells
Authors: Byrne Katelyn T , Kim Samuel I , Arora Charu , Verginadis Ioannis I , Cassella Christopher R , Markosyan Nune , Koumenis Constantinos , Vonderheide Robert H 
Published: January 2023

Abstract
Cytolytic CD8+ T cells are a major mediator of immune-induced tumor rejection, but many patients have primary or acquired resistance to CD8+ T cell immunity. Reduced major histocompatibility complex I (MHC I) expression contributes to resistance, including treatment-refractory pancreatic ductal adenocarcinoma (PDAC). Interrogating interventions in PDAC will reveal novel mechanisms regulating sensitivity to immunotherapy. Using a genetically engineered mouse model of PDAC, we previously reported that agonistic anti-CD40 and dual immune checkpoint blockade (ICB) induces CD8+ and a CD4+ T cell responses mediating tumor rejection. Here, we utilized CRISPR− Cas9 to disrupt the dominant MHC I (H-2Kb) allele expression in PDAC cell lines, precluding direct antigen presentation to CD8+ T cells. Injection of MHC I-deficient tumor clones revealed normal tumor growth at baseline, and significantly delayed tumor growth in response to CD40/ICB in a CD4+ T cell-dependent manner (p &lt; 0.0001 vs. vehicle-treated mice). There was no direct contribution of tumor rejection by CD8+ T cells, despite the generation of a tumor-specific response (24.5% vs. 1.98% in vehicle-treated mice, p&lt;0.003). CD4+ T cells upregulated IFN-γ production (52.3% vs. 11.9% in vehicle-treated mice, p&lt;0.0001), and host IFN-γ expression was required. No changes were observed in the expression of cytotoxic molecules by CD4+ T cells after CD40/ICB, and depletion of myeloid cells did not dampen treatment efficacy. Thus, CD4+ T cells mediated PDAC rejection via non-canonical mechanisms after treatment with CD40/ICB. This study provides critical insight into novel mediators of tumor clearance, and opportunities to exploit CD4 T+ cells as regulators of tumor immunity.
Funded by the Parker Institute for Cancer Immunotherapy.