bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mechanosensory channel Piezo1 is essential in pathogenic T cell-mediated intestinal inflammation
Authors: Choi Sung Hee , Aguilar Alicia , Myers Jay , Kim Byung-Gyu , Eid Saada , Tomchuck Suzanne , Kingsley Daniel , Huang Alex 
Published: January 2023

Abstract
Inflammatory bowel disease (IBD) is a chronic inflammatory disorder in the gastrointestinal tract. CD4+ T cells are especially known to be the main drivers of IBD when they show an elevated level of activation. Elevation of intracellular Ca2+ is one of the key triggering signals for T cell activation. Piezo1 is a mechanosensitive nonselective Ca2+-permeable cation channel, which is broadly expressed in mammalian cells. However, the role of Piezo1 in the pathogenesis of T cell-mediated colitis remains unknown. We have generated T cell-specific Piezo1 knockout (Piezo1fl/flxCD4-cre) mice and observed that loss of Piezo1 in CD4+ T cell increased Th1 and Th17 cell polarization. RNA-sequence analysis of Piezo1fl/flxCD4-cre T cells identified elevated pathogenic Th17 cell pathway, IFN-γ signaling pathways and inflammatory response gene signature compared to that of wild type. These results suggest that Piezo1 controls the inflammatory response of pathogenic T cells. Next, we examined the function of Piezo1 on intestinal inflammation in vivo using acute and chronic colitis mouse model. For the acute colitis mouse model, we used a chemically induced mouse model of colitis using DSS in drinking water. Piezo1fl/flxCD4-cre mice with DSS developed severe colitis compared to Piezo1fl/fl mice with DSS. However, in chronic colitis mouse model, which is the adaptive transfer of naïve CD4+ T cells (CD4+ CD45RBhigh) from Piezo1fl/fl or Piezo1fl/flxCD4cre into Rag1−/− mice, T cells from Piezo1fl/flxCD4cre mice failed to induce colon inflammation, while mice that received T cells from Piezo1fl/fl mice developed severe intestinal inflammation. Thus, our data demonstrate a critical role of Piezo1 in CD4+ T cell-mediated intestinal inflammation.
Supported by R03 CA230840, P30 CA043703, St. Baldrick’s Foundation, Hyundai Hope-on-Wheels Scholar Hope Grant, Andrew McDonough B+ Foundation, Curing Kids Cancer, Center for Pediatric Immunotherapy at Rainbow
Vitamin A regulates phagocytosis by resident macrophages of the small intestine
Authors: Dende Chaitanya , Pendse Mihir , Propheter Daniel , Quinn Gabriella , Hooper Lora V 
Published: January 2023

Abstract
Intestinal Tim4+ CD4+ macrophages are a distinctive macrophage subset that express Tim4, a receptor for phosphatidylserine on dying apoptotic cells, Unlike other macrophage subsets, they do not depend on blood monocytes for their turnover, instead self-maintained in the small intestine. The signal(s) responsible for the self-maintenance and function of Tim4+ CD4+ macrophages is not known. We have discovered that maintenance of the gut resident Tim4+ CD4+ macrophage population depends on dietary vitamin A and its derivative retinoic acid (RA). Retinoic acid receptors, which direct RA-dependent transcription, were required for maintenance of Tim4+ CD4+ macrophages. Chemical blockade of retinoic acid receptor (RAR) signaling and macrophage-specific genetic inactivation of RARs in mice further revealed that macrophage-intrinsic RARα signaling was required for Timd4 expression and maintenance of Tim4+ CD4+ macrophages. Macrophage RARα signaling was furthermore essential for phagocytosis by Tim4+ CD4+ macrophages. Ongoing studies are examining the role of Tim4+ CD4+ macrophages and vitamin A in the clearance of apoptotic intestinal epithelial cells. Our findings reveal that vitamin A provides an essential dietary signal for the maintenance and function of a gut resident macrophage subset.
Supported by Welch foundation grant I-1874
Targeting tonsillar B cells with IL-9 and IL-33 enhances the rhesus macaque humoral response to DNA/protein-based HIV envelope vaccine
Authors: Kobie James , Spencer David , Barnette Philip , Pandey Shilpi , Basu Madhubanti , Sutton William , Rangel-Moreno Javier , Haigwood Nancy Logan , Hessell Ann Jones , Sarkar Sanghita 
Published: January 2023

Abstract
Development of an effective HIV vaccine is dependent on the quantity and quality of HIV Envelope specific antibody at the mucosal sites. Priming of the oral mucosa is a promising approach to generate mucosal antibody at HIV entry sites. Our recent work in mice has demonstrated the ability of IL-9 and IL-33, known regulators of mucosal immunity, to promote robust development of HIV Env-specific IgG; in addition they increase the breadth, magnitude, and durability of the Env-specific IgG response when combined with the DNA/protein HIV Env immunogen platform VC10014, which elicits Tier 2 neutralizing antibody in rhesus macaques. We hypothesized that conditioning the tonsil microenvironment with IL-9 or IL-33 would enable the rapid induction of durable and effective mucosal humoral immunity by the VC10014 DNA/protein HIV vaccine platform. Rhesus macaques were primed with VC10014 gp160 DNA plasmids in the absence or presence of IL-9 or IL-33 intra-tonsillar (IT), followed by intramuscular (IM) boosting with the DNA/protein HIV vaccine regimen. IT immunization with IL-9 or IL33 induced plasma Env-specific IgG that was present as early as week 6, mucosal Env-specific IgG and heterologous plasma neutralizing antibody. IL-33 treated animals exhibited the greatest number of germinal centers (GCs) and largest GCs following the first IM boost whereas IL-9 treated animals showed the greatest number of GCs following the second IM boost in the draining lymph nodes. The difference in kinetics, with IL-33 promoting an earlier B cell response, and IL-9 promoting greater magnitude later, may suggest that IL-9 and IL-33 promote the development of qualitatively different B cells subsets and may have synergistic potential in HIV vaccine development.
Supported by a grant from NIH (R01DE027245).
Microbiota regulation of intestinal inflammation influences colorectal cancer
Authors: Ruiz Daniel Fernando Zegarra , Kim Dasom V , Saldana-Morales Fatima B , Ng Charles , Callaghan Ryann , Uddin Maisha , Chang Lin-Chun , Longman Randy S , Diehl Gretchen E 
Published: January 2023

Abstract
Inflammatory bowel disease (IBD) patients with poorly controlled intestinal inflammation are at an elevated risk for colorectal cancer (CRC). IBD patients exhibit intestinal dysbiosis with expanded proteobacteria such as E. coli. Here, we find that colonization with an E. coli isolated from the intestine of an IBD patient (E. coli 541-15) prevents tumorigenesis in an inflammation-related model of CRC. Colonization increased tumor infiltration of T helper 1 (Th1) cells, cytotoxic T lymphocytes (CTLs), and type 1 innate lymphoid cells (ILC1s) and decreased myeloid derived suppressor cells (MDSCs) and regulatory T cells (Tregs). Prevention of tumorigenesis occurs if colonization takes place before induction of inflammation. Intestinal inflammation in colitis models was ameliorated by E. coli 541-15 and this protection depended on IL-10 production by macrophages and IL-10 signaling to the intestinal epithelium. Colonization with E. coli 541-15 also promotes these IL-10 pathways if colonization occurs after tumorigenesis is established. However, this leads to worse CRC outcome, with increased tumor burden alongside decreased tumor infiltration of Th1 cells, CTLs, and ILC1s and increased MDSCs and Tregs. These results identify activation of an IL-10 signaling loop between immune cells and the intestinal epithelium after E. coli colonization that modulates intestinal inflammation and CRC. Importantly, these pathways can be protective or pathogenic depending on timing of activation.
Supported by 2021 Ludwig Center Basic and Translational Research Award
Migratory type 2 dendritic cells mediate mucosal Th17 response to gut commensal bacteria
Authors: Ngoi Soo M , Yang Yi , Iwanowycz Stephen , Gutierrez Jennifer , Hill Megan , Allen Carter , Chung Dongjun , Liu Bei 
Published: January 2023

Abstract
Dendritic cells (DCs) are professional antigen-presenting cells equipped with MHC-restricted antigens, co-stimulations, and cytokines that effectively prime and differentiate naïve T cells into distinct functional subsets. The immune signals that DCs carry reflect the route of antigen uptake and the innate stimuli they received. In the mucosal tissues, owing to the great variety of foreign antigens and inflammatory cues, DCs are predominantly activated and migratory. The gut commensal bacteria contribute to immune health by educating the immune system at this unique location interfacing with the outside environment. In the small intestine, CD4 Th17 cells are abundant and play a critical role in mucosal defense against bacterial and fungal infections. We identified that the early priming step of commensal-driven Th17 cells is controlled by bona fide Zbtb46-expressing DCs. CCR7-dependent migration of DC2s from the small intestine to the mesenteric lymph nodes is essential for the activation of naïve CD4 T cells. The migratory DC2 population in the MLN are almost exclusively Esam+ cells. Single-cell RNA sequencing highlighted the abundance of co-stimulatory markers (CD40 and OX40) and chemokines (Ccl22 and Cxcl16) on MLN migratory DCs. Further resolution of MLN migratory DC2s revealed that the Th17-polarizing cytokine IL-6 colocalizes with DC2s expressing CD40, Ccl17, and Ccl22. Our study indicates that the mucosal Th17 cell response is regulated by a restricted subset of migratory DC2s in the gut draining lymph nodes.
Supported by NIAID U01AI125859
Interactions Between Aryl Hydrocarbon Receptor and the Microbiome Regulate Intestinal Macrophages in Non Obese Diabetic Mice
Authors: Ehrlich Allison , Malany Keegan , Sandoval Simone , Shulzhenko Natalia , Morgun Andrey 
Published: January 2023

Abstract
Type 1 diabetes (T1D) arises from a complex interaction between genetic and environmental factors. Multiple environmental signals predicted to impact T1D development occur at the intestinal surface, as a result of exposures through ingestion, dietary antigens, and microbial colonization. It is hypothesized that an inappropriate immune response to these exposures precipitates overt T1D. We discovered a novel mechanism by which environmental surveillance interacts with the immune system: immunosuppressive lamina propria (LP) macrophage regulation via the aryl hydrocarbon receptor (AhR). In NOD mice, AhR negatively regulates a macrophage subset (CD11b+ CX3CR1+ MHCIIhi; AhR-MFs) that have a strong positive correlation CD4+ IL10R+ Foxp3+ Tregs in the small intestine. To identify the potential source of AhR ligands that regulate these resident-like macrophages, mice were fed an AhR ligand-deficient diet. These cells were not further regulated by dietary ligands, suggesting that AhR activation originates from ligands produced by the intestinal microbiome. Therefore, germ-free NOD AhR knockout mice were derived and the AhR-MF population was measured. In the absence of the microbiome, both AhR knockout and wild type mice had high levels of AhR-MFs, demonstrating that the microbiome is required for AhR-mediated regulation of LP macrophages. RNAseq analysis of sorted AhR-MFs demonstrate that these cells are antigen presenting but lack an inflammatory gene signature. Currently, single cell sequencing of CD45+ LP cells from AhR knockout NOD mice is being performed to identify how AhR alters macrophage differentiation and predict mechanisms of crosstalk with intestinal CD4+ T cell populations.
Supported by R00 DK117509-03, T32 ES007059
Modeling Inflammatory Immune Cell Recruitment and Response on Human Colon Intestine-Chip
Authors: Carman Christopher , Kanellias Marianne , Ramos Dennis , Maniar Kairav , Saud John , Lucchesi Carolina , Kulkarni Gauri , Apostulou Athanasia , Kujala Ville J , Ewart Lorna 
Published: January 2023

Abstract

Objective
Immune cell recruitment into tissues is an essential step in inflammatory responses. This occurs in a highly tissue- and stimulus-specific manner, which presents a significant challenge to modeling disease and testing therapeutics ex vivo. We previously developed an advanced primary human vascularized Colon Intestine-Chip model and showed that it recapitulates physiologic cell composition, morphology and barrier function. The goal of this study was to test the ability of this system to model inflammatory bowel disease (IBD)-like immune cell reactions ex vivo.

Methods
We perfused primary human peripheral blood mononuclear cells (PBMC) across the vascular channel in untreated ‘resting’ or TNFα/chemokine-treated ‘inflamed’ Colon Intestine-Chips. We analyzed total cell recruitment, inflammatory cytokine secretion and barrier function in the following 24–72 hours.

Results
We show that the perfused PBMC efficiently adhered and transmigrated to the epithelial channel in an inflammation-specific manner. This was followed by an accumulation of proinflammatory cytokines characteristic of IBD (e.g., INFγ, IL1β, IL18), as well as loss of barrier function, the hallmark feature of IBD. We further showed that 1) the recruited cells were strongly enriched in the ‘gut trophic’ α4β7+/CCR9+ PBMC subsets and 2) this recruitment could be blocked with the IBD therapeutic Entyvio, which targets the α4β7-MAdCAM-1 interaction.

Conclusion
Our findings indicate that our Colon Intestine-Chip can model inflammatory immune cell recruitment and in situ immune reactions that reflect key clinical correlates of IBD. This model may prove effective for development of new anti-inflammatory therapeutics for human intestinal diseases.
Supported by Emulate Bio
High-precision micro-wells in polymer for single cell cloning and monitoring
Authors: Schimke Magdalena M 
Published: January 2023

Abstract
STRATEC Consumables is focusing on collaborations in development of smart consumables that enable precise clinical diagnostics, single cell handling and detection of rare proteins out of complex samples. Creation of micro-and nano-features in polymers that allow high-volume replication with great accuracy to ensure reliable results in clincis and laboratories. Micro-wells are supportive to novel approaches in single cell handling, cloning, sequencing and proteomics. STRATEC’s partnership with ALS GmbH from Jena, led to the development and production of nanowell plates for automated screening and recovery of single cells, under the name “CellCelector™”. This unique system combines high-resolution fast fluorescence imaging, sensitive cell detection technology as well as patented robotics for fast recovery of pure and viable cells and is compatible with various upstream cell enrichment approaches.
In order to realize the nanowell plates STRATEC Consumables made use of mastering of high aspect ratio features up to 1:10, followed by injection molding with a high pattern fidelity, assembly to a standard micro-titer plate and surface functionalization to enable the application of ALS. Together, this project shows outstanding results though tight collaboration combining capabilities to innovate in the vibrant field of single cell understanding and applications.
Incorporation of Id3 enhances CAR T cell therapeutic efficacy against solid tumors in an immunocompetent murine model
Authors: Reed Steven James , Tower Samantha J , Dunga Meenakshi , Toumi Ryma , Kalia Vandana , Sarkar Surojit 
Published: January 2023

Abstract
T cell exhaustion in the tumor microenvironment (TME) has hindered the success of T cell-based immunotherapy against multiple types of solid tumors. To combat this exhaustion, we have engineered a CD19-targeted fourth generation CAR construct to express the pro-memory transcription factor Inhibitor of DNA Binding 3 (Id3). Id3 critical for the formation of long-lived memory T cells in viral infection models, and retrovirally-induced expression of Id3 in T cells has been shown to enhance the survival of effector T cells in both acute infection and tumor models.Using an immunocompetent murine model of syngeneic solid tumors expressing truncated human-CD19 (hCD19t), we demonstrate that ectopic expression of Id3 via 4th generation CARs containing Id3 (Id3-CAR) augments CAR T cell therapeutic efficacy against solid tumors. Amongst cells recovered from established solid tumors, Id3-CAR T cells retained a memory-precursor effector cell (MPEC) like phenotype including elevated expression of TCF-1, while the 3rd generation CAR T cells largely displayed a terminally exhausted (TEx) phenotype. Importantly, the inclusion of Id3 did not compromise CAR T cell proliferation or expression of the effector cytokines IFN-γ and TNFα following restimulation. In addition to superior control of primary tumors, the Id3-CAR T cells instilled greater long-term protection from tumor relapse in mice, which cleared primary tumors (6/7) compared to 3rd generation CAR T cells (3/7). Highlighting Id3 as a candidate, these data present incorporation of pro-memory transcription factors as an attractive engineering strategy to intrinsically combat CAR T cell exhaustion in the TME and promote long-term CAR T cell memory and protection from tumor relapse.
Research reported in this publication was supported by the National Cancer Institute of the National Institutes of Health under Award Number F32CA265056, and Pediatric Cancer Research Foundation.
The effect of alloferon on chemosensitivity of pancreatic cancer through the regulation of SLC6A14 expression
Authors: Jo HyeJung , Lee DaHae , Go Cheolhyeon , Kim Yejin , Kang Jae Seung 
Published: January 2023

Abstract
Pancreatic cancer is one of the most malignant solid tumors with a high mortality rate. Although there have been many trials for effective therapies using chemotherapeutic drugs, such as gemcitabine, pancreatic cancer still shows significantly higher mortality than other types of cancer. Therefore, it is needed to examine the effective therapeutic ways to improve conventional therapy for pancreatic cancer. It has been previously reported that cancer cells possess a high rate of glutamine uptake needed for their rapid proliferation. In addition, it has recently been reported that amino acid transporter SLC6A14 is up-regulated in some cancers in close relation to glutamine metabolism. Alloferon is a peptide isolated from the insect immune system and shows anti-viral and anti-inflammatory effects based on its immune modulating function. In addition, its anti-tumor effect was recently discovered, but the related mechanisms are largely unknown. For this reason, I investigated the effect of alloferon on pancreatic cancer cell lines Panc-1 and AsPC-1. When Panc-1 and AsPC-1 were exposed to alloferon for 3 weeks, I found down-regulation of SLC6A14 expression and decreased glutamine uptake. Given that SLC6A14 is involved in tumor progression and survival by promoting glutamine uptake into cancer cells, alloferon could be an effective way with conventional chemotherapeutic drug, gemcitabine. Therefore, alloferon could be a useful adjuvant for an effective therapeutic way with conventional chemotherapeutic drug, gemcitabine.
This study was funded by the NRF of Korea (No.:2017R1A2B2010948, 2017R1A6A3A11032576, 2020R1C1C1009334)
Coupling the immunomodulatory properties of the HDAC6 inhibitor ACY241 with Oxaliplatin promotes robust anti-tumor response in Non-small cell lung cancer
Authors: Adeegbe Dennis O , Bag Arup , Schultz Andrew , Bhimani Saloni , Dominguez William , Cen Ling 
Published: January 2023

Abstract
Although immunotherapy provides durable therapeutic response in a subset of Non-small cell lung cancer (NSCLC) patients, there is still a need for broadly applicable therapeutic strategies. HDAC inhibitors are a promising class of drugs whose immunomodulatory properties are now being appreciated. In the present study, we evaluated the immunomodulatory properties of the HDAC6 inhibitor, Citarinostat (ACY241) on lung tumor immune compartment and its therapeutic potential in combination with Oxaliplatin. Lung adenocarcinoma-bearing mice were treated with ACY241 or vehicle after which immune profiling and RNA-sequencing were conducted in tumor-associated T cells and macrophages. Ex vivo T cell functional studies, tumor growth assessment and survival of tumor-bearing mice treated with ACY241 and/or Oxaliplatin were also conducted. ACY241 treatment promoted increased presence of T and NK cells as well as activation, proliferation, and effector profile of T cells in the lung tumors of treated mice. Furthermore, tumor-associated macrophages exhibited increased expression of MHC and co-stimulatory molecules while expression of inhibitory ligands were reduced. RNA-sequencing of tumor-associated T cells and macrophages revealed significant genomic changes that is consistent with ACY241-mediated enhancement of immune priming. Finally, ACY241 treatment led to significantly enhanced tumor-associated T cell effector functionality in lung cancer-bearing mice and in patient-derived tumors when combined with the chemotherapy drug Oxaliplatin. Collectively, our studies support ACY241 as a promising HDAC6 inhibitor which coupled with Oxaliplatin promotes robust therapeutic outcomes in a pre-clinical model of NSCLC.
High-dimensional single-cell analysis of the immune response in multiple myeloma and profiling of the T cell repertoire in response to immunomodulatory treatment
Authors: Prazich Jack 
Published: January 2023

Abstract
Multiple myeloma is a largely incurable malignancy of plasma cells in the bone marrow. The standard of care for this disease has increasingly included the use of immunomodulatory drugs such as thalidomide and its derivatives. However, high-dimensional analysis of the immune response in multiple myeloma tumors and the resultant response to immunomodulatory treatment is lacking. Here, we present a single-cell multi-omic analysis of the immune response in refractory multiple myeloma patient tumors including profiling of CD8+ T cell antigen specificities. We also analyze changes in the T cell repertoire and gene expression following treatment of pomalidomide. We observe a contraction of the repertoire that includes the expansion of select clones within the repertoire. These results provide a clearer map of the immune response in multiple myeloma and how patients respond to immunomodulatory treatment.
Supported by grants from CRI Lloyd J. Old STAR Program and NIH R33 IMAT 2018
Evaluation of tumor derived Serpina3n in experimental glioblastoma
Authors: Ayasoufi Katayoun , Wolf Delaney , Zheng Jiaying , Tritz Zachariah Peter , Jin Fang , Gulbicki Lauren , Hansen Michael , Johnson Aaron J 
Published: January 2023

Abstract
Glioblastoma (GBM) is an incurable brain cancer which is associated with severe peripheral immune suppression. Using experimental GBM, we recently determined that peripheral immunosuppression involves thymic atrophy, stunted T cell proliferation, and release of circulating factors that inhibit immune responses. RNA sequencing and proteomic studies have recently determined that transcripts and protein levels of Serpina3n are upregulated in the brain and serum of GBM patients and correlates with poor prognosis. We determined Serpina3n is a highly upregulated gene in the brain and the thymus of glioma-bearing mice using RNA sequencing technologies. We therefore sought to determine the role of Serpina3n in peripheral immunosuppression in GL261 glioma harboring wild type and Serpina3n knockout mice. Glioma implantation in Serpina3n knockout mice resulted in similar survival and peripheral immunosuppression compared to controls. Western blotting experiments confirmed that Serpina3n is expressed and secreted by GL261 gliomas isolated from WT and Serpina3n knockout mouse brains. Together this data suggests that tumor-derived, but not host derived, Serpina3n needs to be evaluated as a putative factor associated with peripheral immunosuppression.
Supported by Brains Together for a Cure grant