bluuebunny/crossref_metadata_2025_split huggingface dataset:
The RNA-binding protein Mex3B regulates airway remodeling through modulation of TGFBR3 mRNA stability in chronic rhinosinusitis
Authors: Nan Chen Ao 
Published: August 2023

Abstract

Objective
To explore the pathogenic function of Mex3 RNA binding family member B (Mex3b) in chronic rhinosinusitis(CRS).

Methods
The expression of Mex3b, TGF-β receptor III (TGFBR3), TGF-β2, p-Smad2 were studied by quantitative RT-PCR, immunohistochemistry, western blotting, and ELISA. The binding of MEX3b with TGFBR3 was test by improved RNA immunoprecipitation–coupled high-throughput sequencing (iRIP-Seq).

Results
Compared to control subjects, mNRA and protein expression of Mex3b in nasal epithelial cells from CRSwNP patients are upregulated. And Mex3b mRNA and protein levels in cultured human nasal epithelial cells(HNECs) obtained from control subjects were upregulated by LPS, CpG and IL-1β. Incresed Mex3b downregulates TGFBR3 expression in airway epithelial cells by binding to TGFBR3 mRNA, which results in downregulated level of TGFBR3 in CRS compared to control subjects. Loss of TGFBR3 inhibits TGF-β2 induced gene transcription in nasal epithelial cells. And Mex3b suppresses TGF-β2 signaling activation by downregulating TGFBR3 expression in nasal epithelial cells. Mex3b and TGFBR3 expression levels are correlated with tissue edema in CRSwNP.

Conclusions
The upregulation of Mex3b in CRSwNP suppresses TGF-β2 signaling activation by downregulating TGFBR3 expression in nasal epithelial cells, and thefore regulates tissue remodeling of CRS.
Commensal probiotic provides long-term protection in a murine model of allergic lung disease by altering the host microbiota
Authors: Swartzendruber Julie A , Del Toro Rosalinda Monroy , Incrocci Ryan , Negris Olivia , McGrath Shaina 
Published: August 2023

Abstract
The incidence of allergic diseases is increasing, with house dust mite allergy, afflicting up to 20% of the population in developed countries. House dust mites (HDM) are a common allergen that can be difficult to avoid and triggers allergic reactions, including wheezing and shortness of breath. Bacillus subtilis is a gut-associated commensal probiotic that can prevent murine HDM-induced allergic lung inflammation. We exposed C57Bl/6 mice to B. subtilis via oral gavage and challenged with intranasal HDM for up to 8 weeks. We found that pretreatment with B. subtilis was able to provide protection from eosinophil infiltration, measured in bronchial alveolar lavage fluid (BALF), for 3 weeks. This loss of protection correlated with an increase in the eosinophil chemoattractant CCL24 levels in the BALF and reduced abundance of the bacterial phylum Verrucomicorbiota in collected feces. Additionally, we found that B. subtilis treatment altered the bacterial composition by increasing the abundance of the bacteria phylum Bacteroidetes. These results support B. subtilis as a prophylactic for preventing allergic lung disease and highlights that protection can last up to 3 weeks. This work also expands our understanding of how B. subtilis mediates protection and that in addition to modifying the immune system, it is also altering the host microbiota.
This work is supported by start-up funds and core facility outsourcing funds from Midwestern University.
Lung microbial dysbiosis during early life promotes predisposition to allergic asthma
Authors: Bloodworth Jeffrey C , Hoji Aki , Cook-Mills Joan 
Published: August 2023

Abstract

Background
Allergic asthma is associated with lung microbial dysbiosis. We hypothesized that the microbiome from the lungs of allergy-predisposed mouse pups is sufficient to predispose pups to development of allergy. Allergic inflammation in pups of allergic dams is blocked by α-tocopherol supplementation of the dam during gestation and nursing, but it is unknown whether α-tocopherol alters microbial composition of the lung.

Methodology
Mouse pups were predisposed to allergy by inducing allergy in the mothers. Microbiome was separated from the bronchoalveolar lavage (BAL) of pups of allergic and non-allergic mothers and transferred intranasally at postnatal day 4. 16S rRNA sequencing was performed on the Illumina MiSeq platform.

Results
Compared to pups born to non-allergic dams and dams administered with a-tocopherol supplementation, mouse pups born to allergic dams had elevated abundance of several genera of the γ-Proteobacteria, Firmicutes, and Verrucomicrobia (p&lt;0.05). There was decreased abundance of genera from Bacteroides, Firmicutes, and α-Proteobacteria (p&lt;0.05). Transfer of lung microbiome from neonates of allergic dams to neonates of non-allergic dams was sufficient to confer responsiveness to allergen in the recipient pups. Neonates of allergic dams were not protected from development of allergy by transfer of donor lung microbiome from neonates of non-allergic dams or neonates of αT-supplemented allergic dams. The donor microbiome from pups of allergic mothers colonized the pup lung, suggesting that early life detrimental bacterial species persists.

Conclusion
The lung microbiome from allergy-predisposed pups is dominant and sufficient for enhanced neonate responsiveness to allergen.

Supported by grants from NIH (R01AI127695)
Impact of the LPS from Cyanobacteria Brasilonema sp. on Lung Inflammation and Glucocorticoid Expression
Authors: Swanson-Mungerson Michelle Anne , Del Toro Rosalinda Monroy , Jiang Kallie , Incrocci Ryan , Mayer Alejandro M S , Williams Philip , Swartzendruber Julie A 
Published: August 2023

Abstract
Inhaled glucocorticoids (GCs) are the first-line treatment for asthma. However, patients with asthma characterized by recruitment of neutrophils show poor sensitivity to GC therapy. A potential etiology of this GC-resistant asthma is aerosolized cyanobacterial lipopolysaccharide (LPS) from cyanobacterial blooms. Previously, we demonstrated that inhalation of LPS from Geitlerinema sp. of cyanobacteria induces neutrophilic lung infiltration in mice. Our current data shows that the LPS from another cyanobacteria, Brasilonema sp., also induces lung inflammation after inhalation. Flow cytometric data demonstrate that Brasilonema sp. LPS induces neutrophil infiltration and ELISA identified increases in pro-inflammatory cytokine levels in bronchoalveolar lung fluid after Brasilonema sp. LPS exposure. Furthermore, IHC data demonstrate that mice exposed to Brasilonema sp. LPS show an increased level of lung epithelial GCRβ, a glucocorticoid receptor (GR) analog that inhibits GC activity. Conversely, expression of GCRα, the GR isoform responsible for glucocorticoid sensitivity, remains unchanged. These data help to elucidate the mechanisms of Brasilonema sp. LPS-induced lung-inflammation and potential glucocorticoid resistance, while highlighting the importance of the toxin in promoting human disease near cyanobacterial blooms.
This work is supported by the Kenneth A. Suarez Fellowship and Intramural Funds at Midwestern University.
Probiotic-derived exopolysaccharide interferes with TH2 priming and inhibits allergic inflammation
Authors: Hollinger Maile Kananiokala , Velez Tania Estefania , Zamora-Pineda Jesus , Knight Katherine L , Sperling Anne I 
Published: August 2023

Abstract
Allergic asthma is an increasing burden in industrialized countries, where treatments are limited to targeting symptoms and end-stage effectors rather than root causes. In recent years, the use of prophylactic, inhaled probiotic cocktails has shown promise in preventing allergic asthma. However, the mechanisms by which such probiotics prevent allergic symptoms and the exact bacterial product(s) responsible for probiotic activity remain to be elucidated. In this study, we focused on exopolysaccharide (EPS) from Bacillus subtilis, a ubiquitous soil bacterium that is commonly found as a dietary supplement. We found that in a mouse model of allergic airway disease, EPS treatment, prior to allergic sensitization and challenge, limits lung mucus hypersecretion and eosinophilia. Furthermore, EPS treatment inhibited accumulation of TH2, but not TH1 and TH17 subsets. To determine whether EPS was affecting TH2 cell priming by dendritic cells (DCs), we compared the composition and phenotype of DCs in the lung and mediastinal lymph nodes (medLN) after EPS treatment. Surprisingly, pre-treatment with EPS induced DC maturation as measured by an increase in costimulatory molecules and a decrease in the ability to uptake allergen and migrate to the medLN. DC maturation by EPS was TLR4-dependent, as TLR4 KO DCs did not respond to EPS. Together, our data suggest that EPS inhibits allergic sensitization by inducing DC maturation prior to allergen exposure, thereby leading to decreased TH2 priming and allergic inflammation. Thus, we provide a potential mechanism by which inhalation of a single probiotic-derived molecule can prevent the development of allergic inflammation.
Supported by grants from NIH (3R01 AI125644-05S1, 1F31 HL156659-01A1)
Loss of microRNA-29 in B cells leads to atopic skin phenotype in mice
Authors: Borbet Timothy C , Hines Marcus J , Koralov Sergei B 
Published: August 2023

Abstract
Developing B cells are highly dependent on microRNA (miRNA) regulation. We have previously demonstrated the miR-29 family of miRNAs contributes to the regulation of B cell survival through fine-tuning of PTEN expression, a negative regulator of B cell receptor (BCR) signaling. When the miR-29 alleles are selectively deleted in B cells, this leads to reduced survival which can be rescued by ablating one copy of Pten to restore normal BCR signaling. We observed that mice with B cells lacking miR-29 also develop an inflammatory skin phenotype associated with lesions, leukocyte infiltration into the skin, and persistent pruritis. Nearly 50% of miR-29-deficient animals die within 6 months of birth, and ablation of Pten to restore BCR signaling did not improve the survival rate. This suggests that additional targets of miR-29 in B lymphocytes are involved in promoting this inflammatory skin condition. Epidermal thickening, granulocyte plasma cell infiltration in the skin, and high serum IgE titers suggest that miR-29 may contribute to the regulation of IgE-driven immune responses and that in the absence of this miRNA family, animals develop atopic dermatitis. Our bioinformatic analysis revealed that miR29 is predicted to regulate the expression of the Il4Ra transcript, and in vitro differentiation assays revealed that miR-29-deficient B cells have a higher propensity for IgE class switch recombination in response to IL-4 and/or IL-13 stimulation compared to wild-type control B cells. We conclude that miR-29 is critical in the regulation of both Pten and Il4ra expression in mature B cells thereby influencing the survival and function of peripheral B lymphocytes, which can have profound repercussions for the development of autoimmune disease.
The Bernard Levine Fellowship
Bispecific antibody Armed activated T cells exhibit time-dependent co-expression patterns of inhibitory ligands and gene expression programs
Authors: Barnes Rob , Thakur Archana , Dolatshahi Sepideh , Lum Lawrence 
Published: August 2023

Abstract
Immunotherapy success has been limited in treating solid tumors. Bispecific antibody Armed activated T cell (BATs) therapy activates, expands, and arms patient T cells ex vivo with bispecific antibodies (BiAb) to produce a “drug” that combines non-MHC restricted cytotoxicity of T cells and specificity of BiAb. BATs therapy promises to improve survival in patients with metastatic breast, prostate, and pancreatic cancer without dose limiting toxicities. However, the anti-tumor potency of BATs varies between patients and decreases with time. Understanding the mechanisms behind this decrease of potency will help identify therapeutic combinations and dosing strategies to maximize patient response.
Low effector:target coculture conditions allowing target cell escape from BAT killing were identified. BAT function was measured by anti-tumor cytotoxicity, flow cytometry, and RNA sequencing. Samples were taken at set timepoints to produce a dynamic representation of BAT functional changes in target escape conditions. We observed time-dependent co-expression of inhibitory ligands in BATs, including increased co-expression of PD-1 and LAG3 during the first 24h of coculture, followed by increased co-expression of TIGIT and LAG3 at later timepoints. RNA-seq revealed upregulation of genes associated with T cell dysfunction, including NR4A and checkpoint proteins TIGIT, LAG3, and PD-1, as early as 1 hr in coculture. These findings were integrated into a mechanistic model of BAT-target cell interactions. Our model demonstrated time-dependent emergence of therapeutically targetable inhibitory receptors, suggesting optimizing dosing targets and administration times may improve synergistic immunotherapy combination outcomes.
Examining methotrexate’s safety and efficacy in combination with immune checkpoint inhibitors to control immune related inflammatory arthritis in cancer
Authors: Papadopoulos Theodore , Santinon Francois , Hudson Marie , Del Rincon Sonia Victoria 
Published: August 2023

Abstract
Immune checkpoint inhibitors (ICIs) are associated with high rates of toxic side effects known as immune related adverse events such as inflammatory arthritis (ir-RA). ir-RA is empirically treated with drugs for rheumatoid arthritis such as methotrexate (MTX) and hydroxychloroquine which was recently shown to reduce the anti-tumor benefit of ICIs in mouse models of cancer. We have set out to determine the safety of MTX use in combination with ICIs, its effect on the anti-tumoral immune response and its efficacy in treating ir-RA in a novel arthritis-B16 PD-L1 tumor mouse model. We observed that MTX did not diminish the anti-tumor benefit of ICIs in our melanoma model and significantly altered the intra-tumoral CD8+T cell population by promoting the formation of central memory cells and diminishing PD1 expression. We have identified that MTX acts on neuropilin-1 (NRP1), a receptor identified as a CD8+T cell exhaustion marker promoting terminal exhaustion and limiting memory cell formation in tumors. We have shown that MTX treatment in vitro reduces the expression of NRP1 on stimulated CD8+T cells. We have also advanced in the development of a novel arthritis-tumor model showing that MTX can control and diminish inflammation in the delayed type hypersensitivity arthritis (DTHA) and reduced the frequency of Th1 and Th17 cells in the popliteal lymph nodes of mice, with Th17 cells known to have a crucial role in ir-RA. Together these results show that MTX is safe for use in combination with ICIs in our tumor model and is effective in our DTHA model that mimics the immune profile of ir-RA. With the development of our combined arthritis-tumor model we will be able to assess MTX’s efficacy in controlling arthritis in ICIs treated tumor bearing mice.
Generation of novel anti-tumor chimeric antigen receptors incorporating downstream T-cell signaling proteins
Authors: Angeles Benjamin C , Moreno Taylor , Balagopalan Lakshmi , Yi Jason , Qin Haiying , Alvinez Neriah , Yamane Hide , Lee Mariah , Pallikkuth Sandeep , Tran Andy , McIntire Katherine , Taylor Naomi , Samelson Lawrence E 
Published: August 2023

Abstract
Chimeric antigen receptors (CAR) are molecules with an antibody-derived extracellular domain combined intracellularly with T cell receptor (TCR) signaling proteins. Although CAR-T cells have been a breakthrough clinically, several challenges persist: adverse effects from cytokine secretion, CAR-T cell exhaustion, and limited sensitivity in tumor microenvironments with low density of target antigen (i.e. tumor relapse and solid tumors). While many efforts have focused on identifying new cell surface targets, we have modified CAR intracellular domains. This idea arose from our super-resolution microscopy study that revealed that adapter molecules form clusters distinct from the TCR complex and that full activation necessitates passing a signaling threshold. From these findings, novel Chimeric Adapter Proteins (CAPs) were designed to trigger signaling downstream of the TCRζ chain. CAPs replace the TCRζ with ZAP70 intracellularly and are fused to an extracellular targeting domain. Thus, the potency problem associated with CARs would be ameliorated as CAPs would bypass kinetic proofreading steps defining the signaling threshold and the inhibitory regulation of upstream molecules. Indeed, CAPs exhibited high anti-tumor efficacy, and significantly enhanced long-term in vivo persistence of tumor clearance in leukemia-bearing NSG mice as compared with conventional CD19–28ζ CAR-T. Mechanistically, CAPs were activated in an Lck-independent manner and displayed slower phosphorylation kinetics and a longer duration of signaling compared with 28ζ-CAR. The unique signaling properties of CAPs may therefore be harnessed to improve the in vivo efficacy of T cells engineered to express an anti-tumor chimeric receptor.
Hinging on Success: Leveraging the Power of CAR T-Cell Therapy through In-Silico Modeling of Hinge Length and Epitope Location
Authors: Mirazee Justin M , Aranganathan Akashnathan , Achar Sooraj , Jia Dongya , Chen Xiang , Vani Bodhi , Chien Christopher D , Pouzolles Marie , DeDe Kniya , Youkharibache Philippe , Walters Kylie , Tiwary Pratyush , Altan-Bonnet Grégoire , Taylor Naomi 
Published: August 2023

Abstract
Genetic engineering of T-cells using chimeric antigen receptors (CARs) has shown promise in the treatment of relapsed/refractory B-cell leukemia. However, a significant percentage of patients will eventually relapse, often due to the emergence of tumor cells expressing low levels of the target antigen. Previous research has demonstrated that the hinge domain of the CAR plays a crucial role in regulating the cytotoxic capabilities of these cells. In this study, we assessed the potential to model optimal hinge length for a given single-chain variable fragment (scFv) in relation to the location of the targeted epitope. We designed an array of hinges originating from CD8α and incorporated these hinges into CARs targeting CD19, CD22, CD33, and CD20 antigens. We found that the location of the epitope is critical in determining the optimal hinge; a narrow range of short hinges is more effective against membrane-distal epitopes while long hinges are more effective against membrane-proximal epitopes. To build an in-silico model, we used AlphaFold2 (AF2), reducing the length of the multiple sequence alignment (MSA) clusters to introduce stochasticity and increase the conformational diversity of the structural predictions for the CAR hinges and target proteins. We discovered that CARs with a predicted intermembrane distance of 15nm between the CAR and ligand were particularly effective against antigen-low leukemic cells. Our findings suggest that intermembrane distance can predict CAR responsiveness against a specific epitope, enabling the rapid optimization of CARs by adjusting hinge length. The development of this modeling strategy will facilitate the design of CARs with optimized cytotoxic potential against a wide range of novel targets.
Intramural Research Program of the National Cancer Institute (Bethesda, MD)
Functional and transcriptome analysis of peripheral blood lymphocytes in pancreatic cancer patients before and after chemotherapy
Authors: Brugiapaglia Silvia , Bulfamante Sara , Curcio Claudia , Follia Laura , Spadi Rosella , Cristiano Carmen , Bonello Lisa , Cappello Paola , Novelli Francesco 
Published: August 2023

Abstract
Pancreatic Ductal Adenocarcinoma (PDA) is one of the most aggressive malignancies with a 5-year survival rate of 11% due to its high resistance to chemotherapy (CT).
Understanding how to better induce an effector response following tumor associated antigen (TAA) stimulation is critical to set up an efficient tailored immunotherapy. Peripheral blood mononuclear cells (PBMC) from PDA patients, before and after gemcitabine-based CT, were in vitro stimulated with four TAA (ENO1, FUBP1, K2C8 and G3P) in presence or absence of immune-check point blockade (ICB). Gemcitabine increased the number of TAA recognized by patient T cells, which positively correlated with survival, a high-rate of TAA-induced proliferative responses and high IFN-γ production. By contrast, ICB suppressed TAA-induced proliferative responses before or after CT. Transcriptional analysis revealed that after CT only, responding patients displayed more differentially expressed genes than non-responding patients, suggesting that Gemcitabine modulates gene expression. The clonotypic analysis of ENO1-stimulated T cells from patients showed that TCRβ repertoire was modulated by CT as some clonotypes. These data indicate that Gemcitabine alone, as opposed to ICB, shifted the immunological tone toward a more effector phenotype. The expansion of V-J rearrangements due to TAA stimulation was enhanced by CT, with a stronger reactivity of precise TAA-specific T cell clones after CT, which support the development of precision immunotherapy in selected patient groups, based on CT combined with TAA vaccination.
Supported by grants from AIRC (ID 19931)
Super2-IL-33 armored CAR T cells reshape the tumor microenvironment to universally suppress solid tumor growth
Authors: Ferry Shannon Lynn , Brog Rachel A , Weyburne Emily , Murad Joana , Huang Yina H 
Published: August 2023

Abstract
Tumor associated immunosuppressive cells limit the efficacy of CAR T cells against solid tumors. To alter immunosuppression in the tumor microenvironment, CAR T cells were armored with an IL-2 superkine (Super2) and alarmin IL-33 and adoptively transferred into immunocompetent mice with established primary or metastatic B16F10 melanoma or intradermal MC38 colon carcinoma. We show that Super2-IL-33 CAR T cells universally promoted the control of solid tumors in mice. Analysis of tumor infiltrating lymphocytes (TILs) revealed that Super2 and IL-33 expanded CAR T cells and endogenous lymphocytes cells, resulting in an increased ratio of CD8 effector T cells to regulatory T cells. The expression of the CAR or effector molecules was dispensable for CAR T cell-induced tumor control, underscoring the contribution of endogenous immune cells in mediating therapeutic efficacy. We next evaluated the effects of Super2-IL-33 CAR T cells in bone, liver, and thymus (BLT) humanized mice to assess efficacy against human tumors in an immunocompetent model. BLT mice were intradermally inoculated with Caki1, a human renal cell carcinoma in which non-treated tumors are highly enriched for regulatory T cells. To determine the impact of Super2-IL-33 armored CAR T cells on human tumors and infiltrating immune cells, tumor growth and TILs were analyzed. Super2-IL-33 armored human CAR T cells recruit endogenous immune cells to the tumor that facilitate the suppression of tumor growth. Thus, the Super2-IL-33 cytokine combination may serve as a universal platform capable of overcoming immunosuppression to promote broad anti-tumor immunity.
Supported by grants from NIH (R43-CA271880 and R01-CA254042)