bluuebunny/crossref_metadata_2025_split huggingface dataset:
DRAK2 is a promising target to inhibit GvHD without negatively impacting graft reconstitution after bone marrow transplant
Authors: Duggar Marygrace S , Wehenkel Marie , Vogel Peter , McGargill Maureen Ann 
Published: January 2023

Abstract
Graft versus host disease (GvHD) is the most common adverse event after allogeneic bone marrow transplant (BMT). GvHD and the immunosuppression required to treat it limit the desired graft versus leukemic effect and are associated with significant morbidity and mortality. A treatment that selectively controls the harmful immune response during GvHD, without compromising the response to pathogens or tumors, would be transformative. Previously, we demonstrated that mice deficient in Drak2 (Drak2−/−) are resistant to disease in multiple models of T cell-mediated autoimmunity but maintain effective responses to pathogens and tumors. Since similar immune pathologies mediate GvHD and autoimmunity, we sought to investigate whether loss of Drak2 reduced GvHD, while maintaining effective immunity to pathogens. Thus, we utilized a murine model of acute GvHD (aGvHD) in which lethally irradiated mice received MHC-mismatched bone marrow and purified T cells that were either Drak2+/+ or Drak2−/−. We found that mice given MHC-mismatched bone marrow along with Drak2−/− T cells had less severe aGvHD compared to mice receiving Drak2+/+ T cells, as demonstrated by lower clinical scores. Interestingly, mice given Drak2−/− T cells along with MHC-mismatched bone marrow had increased proportions of regulatory T cells and decreased effector T cells compared to mice given wildtype T cells. These data suggest that Drak2 expression within T cells contributes to GvHD development, and therefore is a promising target in the treatment of GvHD. Ongoing studies are investigating whether the absence of Drak2 influences the effective repopulation of a functional immune system following BMT.
Funding provided by ALSAC and the St. Jude Graduate School of Biomedial Sciences
Seeking the Sertoli cell complement inhibitory signature to prolong transplant survival
Authors: Washburn Rachel , Kaur Gurvinder , Chilton Beverly , Reilly Brian D , Dufour Jannette 
Published: January 2023

Abstract
Clinical transplantation can treat various illnesses, although without the use of toxic immunosuppressants transplanted tissue is rejected by the immune system. Complement, an enzymatic cascade that leads to cytolysis by insertion of the membrane attack complex (MAC), is a significant component in transplant rejection. Interestingly, Sertoli cells (SC) survive long-term after transplantation across immune barriers without immunosuppression. This study used allotransplant (between same species) and xenotransplant (between different species) models to investigate the role of complement inhibition in SC extended survival. Since SC were resistant to in vitro complement killing, mouse or pig SC were transplanted into mice or rats, respectively. Grafts were collected and analyzed for MAC. MAC was not detected on SC transplants, indicating that SC inhibit complement before MAC deposition, possibly through expression of complement inhibitory proteins (CIP). Bioinformatic analyses of mouse or pig SC identified expression of 13 CIP. After expression of two key CIP, CD46 and CD55, were knocked down, SC were killed by complement in vitro. Taken together, these data indicate that the SC complement inhibitory signature is critical for SC graft survival. Understanding SC complement inhibition is important as it could lead to improved transplant survival with a decreased requirement of harsh immunosuppressants, making this procedure more attainable for patients.
Supported by grants from the CH foundation, the South Plains Foundation, and the Mary Lou Clements-Mann Scholarship
Sex-specific T cell behavior drives differential immune responses in mouse glioblastoma models
Authors: Lee Juyeun , Lathia Justin D 
Published: January 2023

Abstract
Glioblastoma (GBM) is the most common primary malignant brain tumor and has a median survival of 12–18 months with current standard-of-care of therapy. GBM sex differences in incidence and overall survival, with males experiencing a higher incidence and a worse prognosis compared to females. However, the role of immune cells in GBM sex differences outside of myeloid cells remains poorly understood. Using a syngeneic mouse GBM model, we recapitulated the sex differences observed in patients, with shortened survival in male hosts compared to female hosts. These findings were not recapitulated in immuno-deficient mouse strains such as NSG and RAG1KO mice, suggesting a role for immune cells, specifically T cells, in GBM sex differences. Flow cytometry analysis of tumor-infiltrating leukocytes revealed more T cells in female tumors, whereas male tumors were enriched in macrophages. Additionally, more T cells in male tumors exhibited high levels of inhibitory receptors, whereas most female T cells were more functional as measured by expression of anti-tumor cytokines such as TNF, IFN-gamma, and granzyme B. A bone marrow chimera model revealed that male T cells retained their phenotype in female hosts, whereas female T cell behavior was affected by the male environment. Furthermore, female T cells are more proliferative, with elevated expression of integrins compared to male T cells. Lastly, we found that males have more progenitor exhausted T cells, which led to better response to anti-PD1 treatment. Collectively, these results suggest that both cell-intrinsic and cell-extrinsic factors regulate T cell activity in a sex-specific manner, providing insights to develop sex-specific therapeutic approaches.
Supported by grant from NIH (P01 CA245705)
Senescent cell-derived extracellular vesicles are critical elements in senescence surveillance by recruiting antigen-presenting cells
Authors: Ziglari Tahereh , Claudio Natalie , Guo Zihan , Pucci Ferdinando 
Published: January 2023

Abstract
Cellular senescence is a tumor suppressor mechanism that arrests cell cycle and initiates immune-mediated removal of senescent cells, a process known as senescence surveillance (SS). Senescent cells interact with their microenvironment through senescence-associated secretory phenotype (SASP). Current knowledge about the SASP has focused on soluble elements like cytokines. Extracellular vesicles (EVs) are membrane-bound, nanometer-sized particles that are emerging as a novel class of SASP factors. Whether EVs play a role in SS is unknown. Elucidating the role of EVs in SS will open new avenues for therapeutically targeting cellular senescence. In order to study if and how EVs contribute to SS, we inhibited EV release from senescent cells using dominant-negative mutant Rab35. This approach preserved secretion of soluble SASP elements while decreasing &gt;95% the release of EVs from senescent cells. C57BL6 mice were challenged with EV-inhibited senescent cells and their numbers at the site of injection as well as immune infiltrates were quantified. Interfering with EV release led to the reduction of antigen-presenting cells including dendritic cells, macrophages, and B cells, which may contribute to the observed persistence of senescent cells. Interestingly, EV-inhibition reverted cellular senescence, promoted proliferation and tumor formation. Proteomic analysis revealed 35 proteins that were exclusively present on senescent cell-derived EVs, compared to those from their non-senescence counterparts. These results suggest that EVs have critical role in SS possibly via signaling to immune cells. Future work will explore whether T cells respond to EV-bound senescent cell antigens.
Integrated Training in Quantitative and Experimental Cancer Systems Biology, 5T32CA254888-02 and CRUK/OHSU A29681
The nucleic acid sensor RIG-I as an inducer of anti-tumor response
Authors: Fraile-Bethencourt Eugenia , Khou Sokchea , Anand Sudarshan 
Published: January 2023

Abstract
The immune microenvironment plays a critical role in tumor progression. Recently, approaches to target Pattern Recognition Receptors (PRRs) have emerged as a promising strategy to enhance immune recognition of tumors. Among the PRRs, those which recognize DNA and RNA in the cells are unique in their ability to respond to both pathogenic and self-nucleic acids that are generated as a result of aberrant cell division/transcription. We previously showed that targeting the DNA sensor three prime exonuclease 1 (TREX1) enhanced immune responses in mouse tumor models. Here we show that the RNA sensor Retinoic acid Induced Gene 1 (RIG-I) acts as a robust immune activator across cancer types. According cBioportal data, expression of RIG-I seems to be a prognostic factor for progression free survival. To functional evaluate RIG-I response in tumor models, we used a hairpin RNA derived from H1N1 sequence. Interestingly, activation of RIG-I increased expression of TREX1 revealing a potential crosstalk between the two pathways. Both breast and colorectal tumor cell lines produce IFN-I after RIG-I activation. Consistently, RNAseq showed upregulation of IFN-I response genes and proinflammatory cytokines across different cell lines. Activation of RIG-I in the tumor cells decreased tumor burden with a concomitant broad cellular and molecular anti-tumor response. Thus, results showed an increase in NK and DC population, while the transcription profile was upregulation of innate immune response genes. Interestingly, tumor growth and NK results were consistent when tumor was implanted in immunocompromised mouse model. Taken together, our findings illustrate how tumor cell RIG-I is a potent inducer of cellular and molecular immune responses.
Diminished noncanonical NF-κB signaling induces colitis-associated colorectal cancer susceptibility upon de-differentiation of epithelial cells
Authors: Morrison Holly , Rowe Audrey , Eden Kristin , Baumgarner Katherine , Brown Stephan L , Holl Eda K , Allen Irving Coy 
Published: January 2023

Abstract
Patients with a medical history of Inflammatory Bowel Disease are predisposed to developing colitis-associated colorectal cancer (CAC), with greater than 20% developing CAC within 30 years of disease onset. CAC is a form of inflammation-induced tumorigenesis, where unregulated inflammation causes cells in the colon to accumulate mutations poising them for neoplasia. We demonstrate that NF-κB inducing kinase (NIK) is a critical regulator of intestinal epithelial cell (IEC) regeneration. We hypothesize that IECs exposed to chronic inflammatory conditions mitigated by dysregulated noncanonical NF-κB signaling are most prone to mutagenesis and cancer-initiating events termed Top-down tumorigenesis. In CAC, NIK depletion results in the accumulation of mature IECs that are increasingly susceptible to mutation and neoplastic transformation. This suggests that terminally differentiated IECs dedifferentiate into a stem-like phenotype. Our work has clinical relevancy, where human CAC biopsy samples have diminished noncanonical NF-κB signaling and NIK (MAP3K14) gene expression. Murine models with NIK deletion in IECs have increased susceptibility to inflammation-induced tumorigenesis following the chemical induction of AOM/DSS to induce colitis. Our mechanistic studies evaluating crypts demonstrate accumulation of mature, non-dividing IECs, which are more susceptible to mutations and malignant transformation. Here, we present a novel role for noncanonical NF-κB signaling in regulating intestinal epithelial cell turnover in protecting against CAC development. Focusing on the key niche signaling interplay following noncanonical NF-κB dysregulation gives greater insight into the CAC tumor microenvironment.
Supported by grants from the iTHRIVE Pilot Translational and Clinical Studies Program, VCOM Center for One Health Research, Virginia Tech Carilion Research Acceleration Program, and the Virginia-Maryland College of Veterinary Medicine.
Egfl6 promotes ovarian cancer progression by inducing the immunosuppressive functions of tumor-infiltrating myeloid cells.
Authors: Cascio Sandra , Sinno Sarah , Bai Shoumei , Coronnello Claudia , Vlad Anda , Buckanovich Ronald J 
Published: January 2023

Abstract
Myeloid-derived suppressor cells (MDSCs) and tumor-associated macrophages (TAMs) are critical negative regulators of immunity in cancer. Understanding factors which regulate these cells could result in the identification of new approaches to enhance anti-tumor immunotherapy. One such factor is epidermal growth factor-like 6 (EGFL6), a secreted factor known to promote cancer stem like cell migration and regulate cancer cell differentiation. We found that mice which overexpress Egfl6 have an increased numbers of granulocytes and monocytes in both the bone marrow and spleen. In vitro and ex-vivo analysis indicated that EGFL6, via binding with beta integrins and activation of SYK and ERK signaling, (i) acts as a chemotactic factor for myeloid cells migration and (ii) promotes their differentiation toward a suppressive state. Suggesting an important role in promoting an immunosuppressive tumor microenvironment (TME), using two syngeneic mouse models of ovarian cancer, we found that expression of Egfl6 in tumor cells resulted in increased accumulation of intra-tumoral MDSCs and TAMs and fewer cytotoxic CD8+ T cells. This was associated with increased tumor growth and shortened animal survival. Gene expression profiling of tumor infiltrating myeloid cells indicated that Egfl6 induced the expression of immunosuppressive factors, including CXCL2, IL-10 and PD-L1. Moreover, in an immune ‘hot’ tumor model, EGFL6 completely inhibited response to a-PD-L-1 therapy. Combined our data show that EGFL6 induces the recruitment of myeloid cells into the ovarian TME and subsequently promotes their immunosuppressive functions. This suggest EGFL6 is a potential novel therapeutic target to enhance response to immune therapy in OvCa patients.
Supported by Ovarian Cancer Research Alliance
Glycoprotein-NMB (GPNMB) is Pro-Tumorigenic in Lymphangioleiomyomatosis (LAM)
Authors: Gibbons Erin , Taya Manisha , Hammes Stephen 
Published: January 2023

Abstract
Lymphangioleiomyomatosis (LAM) is an estrogen-sensitive lung disease found almost exclusively in women. LAM is characterized by the hyperproliferation of smooth muscle cells creating small tumors throughout the lungs, resulting in the formation of large cysts that replace normal alveolar space. Growth of these tumors and progression of the cyst development leads to pulmonary function loss, and sometimes subsequent lung transplantation. However, LAM recurs after transplantation and LAM cells can be found circulating in body fluids suggesting a metastatic nature of the disease. Due to LAM’s estrogen sensitivity, female specificity, and metastatic nature, we previously proposed that LAM cells originate from the uterine myometrium. We therefore designed a uterine-specific TSC2-null mouse model where all of the mice generate uterine tumors characteristic of LAM and half develop lung metastases. Using RNASeq analysis of uterine tissue from this mouse model, when focusing on genes regulated by estrogen and TSC2, we discovered increased expression of melanocytic markers, including Glycoprotein Non-Metastatic Melanoma Protein B (GPNMB), which is known to be oncogenic in other cancers. GPNMB was not only highly expressed in our mouse model, it was also expressed in TSC2-null cell lines, and LAM patient lungs and uteri. Interestingly, serum GPNMB is a potential biomarker, as LAM patients have higher serum GPNMB levels compared to healthy controls. Knocking down GPNMB expression by siRNA decreased proliferation, migration, and invasion in TSC2-null cells. Further, knocking out GPNMB in TSC2-null cells using CRISPR/Cas9 decreased xenograft tumor growth in mice and GPNMB is a potential therapeutic target for LAM.
Supported by grants from NIH (R01 CA193583, T32HL066988), the LAM Foundation, and DOD
Defining myeloid plasticity and heterogeneity in immunotherapy response
Authors: Dinh Huy , Golfinos Athena , Wang Wei , Mergaert Aisha , Lambert Paul 
Published: January 2023

Abstract
Like many cancer types, Head and neck cancers (HNC) have a &lt;20% response rate to immune checkpoint inhibitor (ICI) therapy, making it imperative to learn what causes high treatment resistance. In addition, dissecting the tumor immune heterogeneity and plasticity is crucial to understand and overcome these challenges. Recent scRNA-seq studies have extensively characterized lymphocyte cells in HNC. However, myeloid cells are not well characterized, despite composing many in tumors and exhibiting pro- and anti-tumoral phenotypes. Here we show a comprehensive cross-species characterization of myeloid cells in HNC humans and mouse models and how they influence ICI response. To this end, we generated scRNA-Seq of immune cells in tumors from two HNC mouse models with different responses to ICI. Our comparative analysis identified critical changes in the myeloid compartment in responder and non-responder mice receiving ICI (combined anti-PD1 + anti-CTLA4) therapies. Examples are macrophage and dendritic cell subsets expressing CXCL9 and PDL1, two predictors of ICI response reported in HNC clinical trials and recently published work. Our bioinformatics approaches also defined ligand-receptor cell-cell interactions between myeloid and T cells that influence downstream gene expression networks as potential underlying mechanisms and actionable targets for ICI response. Interestingly, we observed a high concordance between mouse and human myeloid cells from ours and publicly available scRNA-Seq data. Our data and analysis provide the first translational mouse-to-human atlas of myeloid cells in HNC, providing a way to inform biomarkers and immunotherapeutic targets for potential ICI therapy success in HNC and other cancer types.
This research was supported by the University of Wisconsin Head and Neck SPORE grant CEP award (P50DE026787 to H.Q.D)
Cellular avidity as a novel biomarker for candidate selection of cell-based immunotherapies
Authors: Eberlein Jens , Moser Justin , Zhong Zhou , Bailey Keith A , Hoang Song-My , Schultz Aric J , Candelli Andrea , Reijmers Rogier M , Singleterry Will 
Published: January 2023

Abstract
Cellular immunotherapies are increasingly complex in design. Affinity readouts, i.e., the measurements of the strength of single receptor-ligand interactions, are frequently used to assess the binding of chimeric antigen receptors (CARs) or T cell receptors (TCRs) to their ligand. However, affinity is a poor predictor of effector function of the engineered downstream product. Therefore, new tools are needed to evaluate the binding strength of CAR and TCR-transgenic cells to their cognate targets in a more biologically relevant context.
Recent studies demonstrated the utility of measuring cellular avidity as a novel biomarker for identifying and developing potent and safe immunotherapies. Unlike affinity, cellular avidity is driven by the overall strength of dynamic surface interactions between effector cells and their targets by integrating receptor density, the sum of individual affinities, and engagement of the multitude of co-receptors within the immunological synapse.
Here, we show that increased specific avidity, i.e., TCRs with the strongest antigen binding and the lowest background, correlated with improved effector function both in vitro and in vivo. For CAR-T, higher cellular avidity was significantly correlated with improved tumor control in in vivo murine models, but also associated with toxicities in patients, suggesting fine-tuning of cell therapies to the desired avidity for ideal function is needed.
TIM-3+ T regulatory cells alter the lymphoid compartment in cold and Hot solid tumor Model.
Authors: Banerjee Hridesh , Nieves-Rosado Hector M , Murter Benjamin , Kane Lawrence P 
Published: January 2023

Abstract
There is a progressive TIM-3+ T reg increase in solid tumors as the tumor progress. W have found that knocking out the ability of T reg to induce TIM-3 lead to delayed progression of tumors in the TIM-3 conditional knockout mice upon both hot (MC38) and cold (B16F10) syngeneic tumor challenges. We are employing our conditional T reg specific TIM-3 Knockout mouse to study if presence or absence of TIM-3 on T reg cells within these tumor models. Preliminary data has shown that our conditional Tim3 knockout T reg cells are found at a much lower frequency, and have an altered suppressive phenotype within the microenvironment of these tumors. We are currently investigating the other changes in the different subsets of lymphoid compartment in tumors. These data suggest that altering the expression of TIM-3+ on T reg has the potential to act as an effective target to alter tumor microenvironment.
Targeting TIM-3+ T reg may therefore be attractive model to study, along with other combination immunotherapy to increase the success in immunotherapy on patients who develop resistance to primary immunotherapy.
Identification of adjuvants for γδ T cells in response to a novel M. tuberculosis vaccine antigen
Authors: Phelps Kathleen , Xia Mei , Rasi Valerio , Fox Christopher , Petrovsky Nikolai , Carter Darrick , Hoft Daniel 
Published: January 2023

Abstract
γδ T cells are ideal effector cells against infectious diseases and cancer. They reside in blood, lymphoid organs, and mucosa, readily available for host defense. γδ T cells develop memory responses independent of MHC presentation. They perform immune functions such as cytolysis, cytokine production, antigen presentation, dendritic cell maturation, and promotion of B cell and T cell responses. Our lab reported that γδ T cells responsive to M. tuberculosis-specific 6-O-methylglucose-containing lipopolysaccharides (mGLP) inhibit intracellular mycobacterial growth. Preliminary data from NHP studies indicate that vaccination with mGLP and IL-2 greatly reduces Mtb burden post-challenge. IL-2 is not translatable for adjuvant use in humans. Therefore, novel adjuvants for γδ T cells are needed to develop optimal vaccine strategies. We are investigating adjuvant candidates to determine their ability to induce optimal activation and expansion of γδ T cells with mGLP or the phosphoantigen HMBPP. To screen adjuvants, PBMCs are incubated with antigen and adjuvant for 7 days, then analyzed for expansion of γδ T cells. Initially few adjuvants promoted γδ T cell expansion in response to HMBPP and none to mGLP. When matured monocyte-derived DCs were added, multiple adjuvants promoted γδ T cell expansion in response to HMBPP and mGLP stimulation without IL-2. We will determine mechanisms involved in novel adjuvant activity, whether adjuvant and mGLP-expanded γδ T cells inhibit intracellular mycobacterial growth, identify cytokines involved in candidate adjuvant effects, characterize cell surface markers upregulated on DC and γδ T cells, and test the best candidate adjuvant(s) with mGLP in NHP trials.
Supported by Gama Delta R01 AI048391