bluuebunny/crossref_metadata_2025_split huggingface dataset:
Pharmacological activation of protein tyrosine phosphatase SHP-1 ameliorates autoimmune arthritis in mice
Authors: Markovics Adrienn , Toth Daniel M , Glant Tibor T , Mikecz Katalin 
Published: January 2023

Abstract

Background
The Src homology region 2 domain-containing phosphatase-1 (SHP-1) is known to exert negative regulatory effects on immune cell signaling. We observed previously that mice overexpressing an SHP-1 transgene were resistant to the development of autoimmune arthritis and showed decreased immune responses in the cartilage proteoglycan (PG)-induced (PGIA) model of rheumatoid arthritis (RA).
We sought to explore if pharmacological activation of SHP-1 in wildtype (WT) mice prevents or ameliorates arthritis.

Methods
To induce PGIA, mice were immunized with human (PG) in adjuvant. For in vivo SHP-1 activation, mice were treated with regorafenib (BAY 73-4506) per os either before arthritis symptoms developed (preventive treatment) or shortly after the initial signs of arthritis (therapeutic treatment). Regorafenib administration was continued for at least 21 days while arthritis incidence and severity were monitored.

Results
Preventive regorafenib treatment of WT mice significantly decreased arthritis severity (10 or 20 mg/kg) and incidence compared to vehicle treatment. Preventive treatment (10 mg/kg) increased tyrosine phosphatase activity of spleen cells and decreased IFNγ secretion by splenic T cells in vitro. Therapeutic treatment with 15 mg/kg decreased the severity of arthritis and lowered IFNγ production.

Conclusion
Our studies are the first to investigate the role of SHP-1 in autoimmune arthritis. We found that pharmacological activation of SHP-1 in vivo prevented the development of autoimmune arthritis or ameliorated arthritis severity when applied after the disease onset in the PGIA model of RA. Activation of SHP-1 might open a new therapeutic perspective in RA treatment.
CHOP depletion does not prevent gut inflammation in experimental spondyloarthritis
Authors: Navid Fatemeh , Gill Tejpal , Fones Lilah , Brooks Stephen , Colbert Robert A 
Published: January 2023

Abstract
HLA-B27, which predisposes to spondyloarthritis (SpA), can misfold, generate endoplasmic reticulum stress, and activate the unfolded protein response (UPR). The UPR may contribute to SpA pathogenesis by promoting pro-inflammatory cytokine production. CHOP, which is upregulated by the PERK arm of the UPR, is required for excess IL-23 production during UPR-TLR co-activation. Here, we examined whether CHOP deficiency in HLA-B27-transgenic (B27+) rats affects pro-inflammatory cytokine production and gut inflammation.
CHOP-deficient (CHOP-) Lewis rats were generated using CRISPR/CAS9 editing and crossed with B27+ Lewis animals to create B27+CHOP− rats. CHOP expression was not detectable by western blot in CHOP− cells when treated with thapsigargin, to induce UPR and CHOP expression. In contrast to CHOP+ macrophages, CHOP-cells did not express Il23a upon UPR-TLR co-activation. Histological scoring of colon samples from 6-month-old animals showed a trend to increased inflammation in B27+CHOP− compared to B27+CHOP+ rats (8.05±1.11 vs 6.87±1.40; p=0.07). B27-CHOP− rats had no gut inflammation similar to B27-CHOP+ WT rats (scores ≤ 2). RNASeq analysis of colon tissue showed significantly increased expression of 73 genes in B27+CHOP− vs B27+CHOP+ rats, including pro-inflammatory genes like Il1a, Tnf, and Il4r. TOPPGENE analysis revealed that the differentially expressed genes are involved in processes like inflammation, IL-6 production and cytokine secretion. Our data demonstrate that CHOP depletion does not prevent SpA development, but rather it may exacerbate gut inflammation. This suggests that CHOP may play a protective role in HLA-B27 associated disease. This axis warrants further investigation as a therapeutic avenue.
Chimeric antigen receptor signaling confers antitumor activity to human regulatory T cells
Authors: Ferreira Leonardo , Muller Yannick D. , Kaul Anupurna M. , Shaikh Hashim , Guerrero-Moreno Ryan , Yao Lilian E. , Goodman Daniel B. , Bluestone Jeffrey A , Tang Qizhi 
Published: January 2023

Abstract
Regulatory T cells (Tregs) are a subset of T cells dedicated to suppressing immune responses to ensure immune self tolerance. Treg therapy offers the opportunity to treat transplant rejection and autoimmunity without the toxicity associated with immunosuppressive regimens. Conferring antigen specificity to Tregs using a chimeric antigen receptor (CAR) dramatically expands what targets can be pursued using Treg therapies. However, the consequences of CAR-mediated signaling for human Treg biology remain poorly understood. To address this, we generated anti-CD19 CAR Tregs with a tandem CD28-TCRζ intracellular domain. Upon in vitro co-incubation with CD19-expressing cells, CAR Tregs upregulated activation markers, proliferated, secreted IL-10, and suppressed T cell proliferation, while maintaining high FOXP3 expression and a demethylated TSDR. In humanized mice harboring CD19+ tumor cells, CAR Tregs suppressed CAR T cell proliferation. Strikingly, CAR Tregs also suppressed tumor growth, even in the absence of CAR T cells. This phenomenon was observed across three tumor cell types (B-cell leukemia, myeloid leukemia, and epithelial carcinoma) and two routes of delivery (intravenous and subcutaneous). Real-time monitoring of tumor cell survival and proliferation in vitro confirmed that CAR Tregs suppress tumor cell growth. Interestingly, single-cell cytokine analysis revealed that CAR-mediated activation of Tregs leads to higher production of IFN-γ, TNF-α, perforin, and granzyme B than anti-CD3/28 stimulation. CRISPR/Cas9-mediated ablation of granzyme B in CAR Tregs diminished their antitumor activity in vivo. These results demonstrate that CAR Treg cytotoxicity is an important concern for safe clinical translation.
Novel mRNA-based Therapeutics to Modulate T and B Cell Signaling and Function
Authors: Kenney Laurie Lea , Fu Ying , Stein Paul , Huang Eric 
Published: January 2023

Abstract
Autoimmunity is considered a T cell driven disease and in many cases B cells contribute to the underlying pathology. Many of the current therapeutics used to treat autoimmunity and organ transplantation are broadly immunosuppressive and can result in increased susceptibility to infection and cancer. We have designed several mRNA-encoded chimeric polypeptides containing an inhibitory motif and a location motif that are expected to disrupt signaling downstream of the T and B cell receptor (T/BCR) and blunt activation. We found that lipid nanoparticle (LNP)-encapsulated mRNA encoding TCR disruptors (Immune LNP-TCRi) containing either a constitutively active CSK (CA.Csk), ITIM, or SHIP1 inhibitory motif reduced in vitro proliferation and cytokine production in primary T cells. In an in vivo xeno-GVHD model Immune LNP-TCRi significantly delayed T cell-mediated mortality. BCR disruptors were engineered by creating mRNA encoding chimeric proteins containing components of the BCR complex (CD79a, CD79b and CD19) and either the CD22 intracellular ITIM or SHIP1 inhibitory motif. Encapsulated in Immune LNP, these BCR disruptors downregulated B cell activation as measured by phosphorylation of Syk, suppressed BCR-induced IgM/G, IL6 and IL10 secretion, as well as reduced B cell in vitro proliferation. In NSG mice engrafted with healthy human PBMC, Immune LNP-BCRi also suppressed humoral immunity against tetanus toxoid and reduced serum IgM and IgG levels. Unlike corticosteroids and anti-CD20 Ab, co-engagement of chimeric T/BCR disruptors did not significant decrease the number of lymphocytes. Thus, amplification of T/BCR disruptor pathway may represent a novel therapeutic approach for autoimmune diseases.
Combining tolerogenic agents with dexamethasone synergistically inhibits innate inflammatory responses
Authors: Genito Christopher J , Stiepel Rebeca T , Batty Cole J , Bachelder Eric M , Ainslie Kristy M 
Published: January 2023

Abstract
Excessive activation of innate immune cells drives many diverse inflammatory human diseases, such as septic shock and autoimmunity. Many modern therapies employ immunosuppressants and other tolerogenic agents (TAs) to treat inflammation through inhibiting inflammatory cytokines and other effector molecules secreted from innate immune cells. TAs carry side effects due to general immunosuppression and off-target effects. One alternative is TA combination therapy, which could reduce side-effects through dose sparing and may improve therapeutic efficacy. To this end, multiple commonly used TAs were screened for synergistic inhibition of inflammation. Screened TAs included immunosuppressants and steroids used in the clinical treatment of allergy, arthritis, and autoimmune disease, as well as candidate drugs in preclinical testing. Anti-inflammatory potential was determined by inhibition of reactive oxygen species from activated macrophages in culture. Synergistic inhibition was calculated mathematically for TAs combined in various ratios. At least 4 synergistic hits were obtained from the screen, with minimum combination indices ranging from 0.5 (good synergy) to 0.04 (extreme synergy). All other tested TAs showed additive improvement in combination. Each synergistic combination included dexamethasone (DXM). Using DXM in combination with an otherwise ineffective TA may have reduced serum levels of inflammatory cytokines in a mouse model of septic shock. We conclude that using DXM in combination with other TAs may improve the inhibition of macrophage inflammation in multiple clinical applications.
Antigen-specific B cell depletion with desmoglein 3 chimeric autoantibody receptor T cells (DSG3-CAART) for targeted therapy of mucosal pemphigus vulgaris
Authors: Lundgren Daniel Kjell , Lee Jinmin , Mao Xuming , Manfredo-Vieira Silvio , Wang Baomei , Nunez-Cruz Selene , Williams Erik , Fraietta Joseph , Milone Michael , Payne Aimee 
Published: January 2023

Abstract
Chimeric autoantibody receptors (CAARs) direct T cells to kill autoantigen-specific B cells. We previously established proof-of-concept for the CAAR approach using gene-engineered T cells expressing CAARs comprising DSG3, the autoantigen in the autoimmune blistering disease mucosal pemphigus vulgaris, fused to CD137-CD3ζ signaling domains. Here, we present results of preclinical studies that supported the DSG3-CAART investigational new drug application.
DSG3-CAART demonstrated activity related to dose in a passive transfer polyclonal anti-DSG3 hybridoma mouse model of mucosal pemphigus vulgaris, resulting in reversal of the rising serum anti-DSG3 antibody titers, target cell burden, and IgG deposition in epithelial tissues, as well as increased DSG3-CAART engraftment relative to dose. We further evaluated DSG3-CAART efficacy in a novel exploratory active immune mouse model with physiologic levels of circulating anti-DSG3 IgG, which demonstrated decreased antibody responses against pathogenic DSG3 epitopes and autoantibody binding to epithelial tissues, leading to clinical and histologic resolution of blisters. High-throughput screening of a membrane protein array with soluble DSG3 CAAR did not identify verifiable off-target toxicities.
These preclinical studies have enabled a first-in-human clinical trial of DSG3-CAART in mucosal pemphigus vulgaris and provide a foundation that may inform the preclinical development of future CAAR T cell therapies for antigen-specific B cell depletion in other autoantibody-mediated diseases.
JMJD3-deficient impair CD4 T cell trafficking in autoimmune disease models and adoptive immunotherapy models by targeting cytoskeleton genes
Authors: Fu Chuntang , Xing Changsheng , Yin Bingnan , Chu Junjun , Duan Tianhao , Liu Xin , Wang Rongfu 
Published: January 2023

Abstract
Histone H3K27 demethylase, JMJD3 plays a critical role in gene expression and T-cell differentiation. However, the role and mechanisms of JMJD3 in T cell trafficking remain poorly understood. Here we show that JMJD3 deficiency in CD4+ T cells resulted in T cell trafficking deficiency autoimmune disease models and adoptive immunotherapy models. We identified PDLIM4 and S1P1 as significantly down-regulated target genes in JMJD3-deficient CD4+ T cells by gene profiling and ChIP-seq analyses. The further study showed that PDLIM4 regulate actin cytoskeleton, thus serving as a key regulator of T cell trafficking. These results demonstrate that manipulating JMJD3 expression can benefit autoimmune disease treatment and generate better T cell grafts for adoptive immunotherapy.
Use of CART cells to selectively target autoantigen-specific T cells for the treatment of autoimmune diabetes
Authors: Yu Hua , Bettini Maria , Ellis Gavin , Riley James L , Collins Jesse , Preston-Hurlburt Paula , Korah Maria , Mallone Roberto , Deng Songyan , Wang Xiaoli , Fremont Daved H. , Spiegel David , Cresswell Peter , Herold Kevan C 
Published: January 2023

Abstract
Previous clinical trials using biologics-based broad-spectrum T cell- and B cell-depleting molecules for the treatment of autoimmune diabetes have shown promising, yet mixed, results. Their varied extent of success may be due to their non-specific action and failure to permanently and completely remove the pathogenic subpopulations. As CD8+ T cells, the most dominant cell type in human insulitis, are thought to be the primary mediator of β-cells damage, we thus designed a strategy by adapting chimeric antigen receptor engineered T (CART) technology to directly target these pathogenic T cells.
The newly generated CAR construct maintains original transmembrane and intracellular components, while the extracellular scFv antigen-binding domain was replaced with HLA-A2/β2-microglobulin (B2M) complex that is linked with either diabetes-associated immunodominant peptide zinc transporter 8(ZnT8)186–194 or negative control peptide HIV Gag77–85. We have shown that HLA-A2/B2M complexes were correctly folded and presenting right peptide epitopes on CART cells. The CAR signaling was also sustained, as the expression levels of CD25 and CD69 were significantly elevated only on CAR-transduced Jurkat cells presenting ZnT8186–194 peptide co-cultured with a T-cell line that expresses TCRs recognizing the same peptide. To determine the killing ability of CART cells, we further engineered primary human T cells to express our CAR construct and demonstrated that CART cells can selectively deplete human antigen-specific CD8+ T cells in vitro. Therefore, our results have provided proof-of-principle for the development of a novel immunotherapy for disease treatment.
Identification of T cells specific for DNAJB1-PRKACA fusion neoantigens in fibrolamellar carcinoma
Authors: Kirk Allison M , Crawford Jeremy Chase , Zamora Anthony E , Allen Kaity , Zhang Xiaoyu , Strome Scott E , Thomas Paul G 
Published: January 2023

Abstract
Fibrolamellar carcinoma (FLC) is a rare liver tumor with poor outcomes and for which novel therapies are urgently needed. A potential therapeutic vulnerability in FLC is a gene fusion between DNAJB1 and PRKACA that is expressed in all FLC tumors and has been shown to drive FLC tumorigenesis. Genetic mutations that alter amino acid sequence, including gene fusions, can be presented as neoantigens by MHC class I molecules on tumor cells and thereby elicit anti-tumor CD8 T cell responses. We hypothesize that DNAJB1-PRKACA fusion neoantigens can elicit T cell responses in FLC patients and serve as targets for T cell-based immunotherapies for FLC. To test this hypothesis, T cells isolated and expanded from an FLC patient tumor were stimulated with DNAJB1-PRKACA fusion peptides, which reproducibly induced cytokine production measured by intracellular cytokine staining. Single cell gene expression and T cell receptor (TCR) repertoire analysis of patient tumors using the 10X Genomics platform identified clonally expanded TCR clonotypes that were associated with expression of cytokines and activation markers in response to fusion peptide stimulation. These TCRs were cloned and expressed in Jurkat 76 cells to test their fusion specificity and their ability to mediate in vitro functional responses to fusion neoantigens. When stimulated with their cognate fusion neoepitope, TCR-expressing cells produced cytokines, expressed the early activation marker CD69, and downregulated surface expression of TCR and CD3. These studies have therefore identified T cell responses against the DNAJB1-PRKACA fusion for the first time, as well as identified DNAJB1-PRKACA-specific TCR clonotypes that could be used to develop novel immunotherapies for FLC.
Treatment of CEA-positive solid tumors with anti-CEA chimeric antigen receptor T-cells in CEA transgenic mice
Authors: Cha Seung , Yazaki Paul J , Brown Christine , Shively John 
Published: January 2023

Abstract
Chimeric antigen receptors (CARs) combine specificity of antibody to target antigen on cancer cells with the potent cytotoxic activity of T-cells without the need for MHC recognition. Chimeric antigen receptor (CAR) T-cells that were designed to express the single chain variable fragment binding domain of human anti-CEA antibody BW431/26 on mouse splenocytes were able to reduce CEA-expressing pancreatic adenocarcinoma tumor in CEA-transgenic (CEAtg) mice without significant off-target effects (Chmielewski et al., 2012).
In this study, an anti-CEA CAR construct derived from murine anti-CEA antibody (T84.66) expressed on human T-cells targeted CEA+ colon carcinoma cells (LS174T/CEA and MC38CEA) with high specificity in vitro. When co-incubated with CEA-positive or CEA-negative mouse cancer cells, anti-CEA CAR expressed on mouse T-cells specifically targeted CEA-positive cancer cells in vitro. Compared to mock T-cells control, anti-CEA CAR T-cells injected intravenously delayed subcutaneous MC38CEA tumor growth in CEAtg mice. Lymphodepletion via cyclophosphamide injected before anti-CEA T-cells therapy further delayed subcutaneous MC38CEA tumor growth in CEAtg mice. To improve the therapeutic potential of anti-CEA CAR T-cells, IL2 was conjugated to a humanized anti-CEA antibody to form an immunocytokine (ICK). The ICK was injected interperitoneally after anti-CEA CAR T-cells into lymphodepleted CEAtg mice. A single injection of the ICK delayed subcutaneous MC38CEA tumor growth even further. Four injections of ICK after anti-CEA CAR T-cells eradicated subcutaneous MC38CEA tumors. These data show the therapeutic potential of anti-CEA CAR T-cells to target CEA-positive tumors.
Immunotherapy for solid tumors: Chimeric antigen receptor (CAR) T-cells and the tumor microenvironment
Authors: Nwanze Chiadika , Kratzmeier Sabina , Kaplan Rosandra 
Published: January 2023

Abstract
Patients with solid tumors continue to experience decreased survival rates, with metastasis being the leading cause of mortality in these patients. This has prompted the development of novel immunotherapies, with cellular immunotherapy giving rise to the development of Chimeric Antigen Receptor (CAR) T-cells, an approach combining specificity for tumor antigens and cytotoxicity independent of major histocompatibility complex molecule co-stimulation. This modality has proven efficacious in Acute Lymphoblastic Leukemia and B-cell Lymphoma. However, progress has remained limited in solid tumors despite targeting of various antigens, one of such being disialoganglioside (GD2) which has proven a viable target in neuroblastoma. This study investigates the efficacy of GD2 as an antigen target for CAR T-cell therapy in osteosarcoma. Surface flow cytometric analysis revealed the presence of GD2 on human 143B and Hu09H3 osteosarcoma cell lines. Of note, the tumor microenvironment offers an additional potential therapeutic target, as we observed increases in myeloid-derived suppressor cells alongside decreases in CD4+ T cells in primary tumor and metastatic tissues. In summary, we report the utility of GD2 as a target for cellular immunotherapy in osteosarcoma and the role of the tumor microenvironment in anti-tumor therapeutic modalities, a therapeutic combination that could be applicable to optimizing potent therapeutic modalities for solid tumors.
THERAPEUTIC EFFECTS OF TRAIL-SECRETING MESENCHYMAL STEM CELLS WITH FOCUSED ULTRASOUND-INDUCED TEMPORARILY OPENING OF BLOOD-BRAIN BARRIERS IN THE BRAIN TUMOR
Authors: Park Sang In , Jung Ho Yong , Lee Eui-Jin , Maeng Lee-So 
Published: January 2023

Abstract
Mesenchymal stem cell (MSC)-based gene therapy is a promising tool for the treatment of various neurological disease including brain tumor. The tumor necrosis factor-related apoptotic ligand (TRAIL) is believed to have promise as a cancer the therapy. For stem cell therapy in the brain tumor, therapy efficiency of stem cells by non-invasive method is extremely limited due to blood brain barrier (BBB). Noninvasive pulsed focused ultrasound (FUS) can temporarily opening the BBB of specific areas in the brain. Herein, we investigated the tumor-targeted BBB temporarily opening by FUS and therapeutic efficiency of NIR fluorescence-labeled MSCs-TRAIL by intravenous injection in brain tumor. In addition, we observed the inflammatory changes of microglia cells by FUS using PET-CT and histological analysis. As a results, In vivo survival experiments showed that FUS treated-MSC-TRAIL group has greater therapeutic efficacy than FUS untreated-groups. The number of MSCs accumulated specifically site in the tumor region. MSC migration toward FUS treated-tumor site greater compared to the FUS untreated-tumors. Furthermore, the changes of microglia activity by FUS was no difference between the other groups. Collectively, these results suggest that therapeutic efficacy of MSC-TRAIL increases by temporarily opening the BBB using the FUS, which may be a more useful strategy for cancer therapy.
Lactate is critical for cytokines production of T cell after TCR activation
Authors: Zhang Yaguang , Wen Jing , Sun Bing 
Published: January 2023

Abstract
Glycolysis is considered to be a key regulator of T cell maturation and function. After TCR activation, T cell accelerates the metabolic process of glycolysis, which produces a large amount of lactic acid. Because the dissociation of lactic acid in solution is effectively complete and there is a strong systemic acid-base equilibrium in our body, lactate exists in the main form of lactic acid ion after TCR activation. This neutral environment with a high concentration of lactic acid ion gives us a hint that T cells may recycle this free lactic acid ion for energy recovery or other vital processes. In this study, we found that lactate as the end-product of glycolysis is critical for T cell cytokines production, especially in T cell-dependent tumor immunology. In in vitro T cell activation, the addition of excess sodium lactate promoted more cytokines production (such as IL-2/TNF) than sodium chloride or sodium pyruvate, whose difference depended on TCR/CD3 activation, but not CD28. Further work showed that those “up-regulated” cytokines were owing to post-transcription, which was regulated by enhanced glycolysis after TCR activation with sodium lactate. This enhanced glycolysis attenuated GAPDH binding ability to cytokines mRNA, which permitted more cytokine protein translation. In vivo study showed that sodium lactate treatment could inhibit tumor growth in the MC38 tumor model with more cytokines production from T cells compared to sodium chloride treatment group. Taken together, our finding indicates a new regulatory mechanism of glycolysis in T cell function.