bluuebunny/crossref_metadata_2025_split huggingface dataset:
Bone marrow derived dendritic cells modified by lentiviral-mediated RelB shRNA possess tolerogenic phenotype and functions on lupus splenic lymphocytes
Authors: Mok Mo Yin , Wu Haijing , Lo Yi 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is an autoimmune disease that is characterized by high morbidity and mortality and remains challenging in treatment. Dendritic cells (DCs) have been shown to participate in the initiation and perpetuation of lupus pathogenesis. DCs that can induce tolerogenicity appear as potential cell-based therapy in this condition. In this study, we examined the in vitro tolerogenic properties of bone-marrow derived DCs (BMDCs) in the murine lupus setting. We used lentiviral transduction of RelB-silencing shRNA to modify expression of RelB, a key transcription factor regulating DC maturation, in BMDCs from MRL/MpJ mice. Tolerogenic properties of RelB-modified DCs were compared to scrambled control (SC)-modified DCs. RelB expression was found to be significantly reduced in RelB-modified DCs derived from MRL/MpJ mice, wild type of the same genetic background as MRL/lpr lupus-prone mice. These MRL/MpJ RelB-modified DCs displayed semi-mature phenotype with expression of lower levels of co-stimulatory molecules compared to SC-modified DCs. RelB-modified DCs were found to be low producer of IL-12p70, can induce hyporesponsiveness of splenic T cells from MRL/MpJ and lupus-prone MRL/lpr mice. Furthermore, they downregulated IFN-γ expression and induced IL-10 producing T cells in MRL/MpJ splenocytes, and attenuated IFN-γ and IL-17 expression in MRL/lpr splenic CD4+ lymphocytes. Splenocytes primed by RelB-modified DCs demonstrated antigen-specific suppressive effect on allogeneic splenocytes. In conclusions, RelB-silencing in DCs generates DCs of tolerogenic properties with immunomodulatory function and appears as potential option of cell-targeted therapy.
WX486, a novel inhibitor of BTK, demonstrated dose dependent inhibition of arthritis in mouse CIA and rat PG-PS models
Authors: Lin Jian , Chan Chichung , Wu Chengde , Shen Chunli , Han Shuhua , Li Xin , Guo Tingting , Chen Shuhui , Xiao Qiang , Huang Lu , Chen Hailiang , Tu Ronghua , Xu Yong , Wang Xuehai 
Published: January 2023

Abstract
Bruton’s Tyrosine Kinase (BTK) plays a crucial role in B cell differentiation and proliferation, making it an attractive target to treat inflammatory and autoimmune diseases. A small molecule BTK inhibitor, WX486, showed a high potency and selectivity, with an in vitro IC50 of 2.1 and 59nM in biochemical and cellular assays, respectively. In vivo anti-arthritis effect of WX486 was evaluated in two animal models, a mouse Collagen-Induced Arthritis (CIA) model and a rat peptidoglycan-polysaccharide (PG-PS) induced arthritis model. In both models, the administration of WX486 decreased ankle swelling and reduced morphologic pathology in a dose-dependent manner. In mouse CIA model, the inhibition of WX486 was 65.46%, 82.36%, 83.91% at 1, 3, 10mpk, respectively. In rat PG-PS model, WX486’s inhibition effect was 47.7%, 64.7%, 77.3% at 1, 3, 10mpk. In addition, WX486 at 3mpk demonstrated superior inhibition of arthritis over CC-292, a BTK inhibitor currently in phase II clinical trials for rheumatoid arthritis (RA), in both animal models.
Discovery of WX486 as a novel, potent, selective, and orally bioavailable irreversible inhibitor of BTK
Authors: Wu Chengde , Shen Chunli , Zhu Yuchuan , Hu Guoping , Lin Jian , Chen Shuhui , Xiao Qiang , Huang Lu , Chen Hailiang , Tu Ronghua , Xu Yong , Wang Xuehai 
Published: January 2023

Abstract
Bruton’s tyrosine kinase (BTK) has emerged as an attractive target for therapeutic intervention in human malignancies and autoimmune disorders, such as rheumatoid arthritis. Compounds were designed and evaluated in CADD based on published crystal structure. Selected compounds were synthesized and evaluated by BTK enzymatic assay. A chemical series where the central ring of ibrutinib was replaced by a bioisosteric motif showed strong enzyme inhibition activity. Further SAR of terminal aryl ring and Michael acceptor in terms of enzymatic activity, DMPK, hERG activity, and solubility was conducted. As a result, WX486 was identified as a potent (IC50=2.1 nM), selective (200~5000-fold over other 24 kinases), orally bioavailable (F=26.2%, 42.7%, 40.8% for mice, rats, and dogs, respectively), and soluble (solubility=266.40, 214.47, 209.01 μM at pH 2.5, 6.5, 7.4 respectively) preclinical candidate.
MyD88-dependent PKD1 is essential for development of polyarthritis spontaneously occurring in IL-1R antagonist-deficient mice.
Authors: Yi Ae-Kyung , Yoon Tae Won , Cho Hongsik , Park Jeoung-Eun , Hasty Karen A , Stuart John M 
Published: January 2023

Abstract
IL-1 receptor antagonist-deficient (IL-1rn−/−) mice develop spontaneous polyarthritis that mimics human rheumatoid arthritis. We investigated whether a Toll-like receptor (TLR)/IL-1R signaling adaptor MyD88 and its downstream protein kinase D1 (PKD1) play a role in the development and progression of arthritis in IL-1rn−/− mice. Serum cytokine levels, and macrophage accumulation, matrix metalloproteinase activities and cartilage damage in joints were significantly lowered in MyD88−/−IL-1rn−/− mice compared to MyD88+/+ IL-1rn−/− mice. Development of arthritis was completely inhibited in MyD88−/− IL-1rn−/− mice, indicating an absolute requirement for MyD88 in the development of arthritis in IL-1rn−/− mice. Because PKD1 is essential for proinflammatory responses mediated by MyD88-dependent TLRs, we further investigated the contribution of PKD1 to the development of arthritis in IL-1rn−/− mice. Suppression of PKD1 expression in vitro resulted in inhibition of TLR-mediated cytokine production in IL-1rn−/− macrophages. Daily treatment with a PKD inhibitor substantially reduced the incidence and severity of arthritis in IL-1rn−/− mice. In addition, deletion of the PKD1 gene in IL-1rn−/− mice using a tamoxifen-inducible system significantly delayed development of arthritis and reduced the incidence and severity of arthritis in IL-1rn−/− mice. Our findings demonstrated that MyD88 and PKD1 are necessary for development of arthritis in IL-1rn−/− mice. This also implies that PKD1 might be one of the key factors that modulate proinflammatory responses in rheumatoid arthritis, and may serve as a therapeutic target.
CancerNet: Molecular profiling of tumor microenvironment for biomarker discovery
Authors: Chenchik Alex , Makhanov Mikhail , Komarov Andrey , Sastry Sunitha , Frangou Costa 
Published: January 2023

Abstract
Human carcinomas consist of a complex mixture of neoplastic epithelial cells, endothelial cells, fibroblasts, myofibroblasts, and immune cells, which collectively form the tumor stroma. These cells are implicated in disease progression, metastasis and drug resistance. The clinical importance of tumor immune infiltrates has been an emerging area in triple negative breast cancer (TNBC) research where increased immune infiltrate predicts both response to chemotherapy and improved survival. Quantitative molecular profiles of tumor-associated normal cells may provide important insights into tumor biology and facilitate the discovery of new biomarkers and therapeutic targets. To this end, we have developed CancerNet, a comprehensive targeted RNA-Seq cancer immuno-panel that profiles ~2,000 genes and distinguishes 37 human hematopoietic cell phenotypes, (including naïve and memory B cells, seven T-cell types, dendritic cells, plasma cells, natural killer (NK) cells and myeloid subsets), genes involved in checkpoint blockade and immunotherapy biomarkers, immune cell activation and canonical immune pathway genes for both innate adaptive and humoral immune responses. In this study, we present an assay that characterizes the cellular composition of the immune/stromal/cancer cell tumor microenvironment and determines the activation status of infiltrating immune cells in primary tumor tissues from TNBC patients. Preliminary studies demonstrate the assay’s unparalleled specificity and sensitivity resulting in better detection of low abundance mRNA transcripts as well as an improved cost-effectiveness for high-throughput clinical Next-Gen applications.
Age effects on tumor B7-H1 and related immune therapies in an aggressive model of ovarian cancer.
Authors: Hurez Vincent , Padron Alvaro Souto , Drerup Justin Michael , Liu Aijie , Curiel Tyler J 
Published: January 2023

Abstract
Age is the greatest cancer risk factor, but contributions of aging to tumor intrinsic properties and immunotherapy responses are poorly understood as most preclinical studies are in young hosts. We assessed age effects on tumor properties, progression, immunity and response to immunotherapies in an aggressive ID8 ovarian cancer line (ID8agg) that is poorly immunogenic and fatal in ~5 weeks. Young (3 mos) and aged (24 mos) BL6 females were injected intraperitoneally with 4×106 syngeneic ID8agg cells. In young mice, ID8agg ascites had more regulatory T cells (Tregs) and myeloid suppressor cells (MDSCs). Young mice gained 30% weight from ascites ~30 days after ID8agg challenge and formed 1–2 large, discrete peritoneal tumor masses. Aged mice tended to lose weight although ascites volumes were comparable to young, suggesting cachexia, and formed multiple small peritoneal nodules. Aged ascites had more tumor cells, Tregs, monocytic MDSCs and PD-1+ T cells versus young. Strikingly, tumor cells and MDSCs in aged ascites upregulated the immune checkpoint molecule B7-H1 greater than in young ascites. The combination of depleting Tregs and MDSCs with denileukin diftitox (DD), an FDA-approved IL-2/diphtheria toxin + αGr-1 temporarily slowed tumor progression in aged mice with increased IFNγ+ CD8 T cells, consistent with our prior B16 data. Treg depletion with either DD or αCD25 alone, or plus αCTLA4 did not significantly prevent ID8agg progression in young or aged mice. In young hosts, αB7-H1 slowed tumor growth and increased survival, with both clinical outcomes potentiated by adding αCTLA4. αCTLA4 + αB7-H1 effects in aged hosts are under study. Thus, we identified major age effects on tumor progression, and immune and immunotherapy responses.
DEFINING THE IMMUNE MICROENVIRONMENT IN PATIENTS WITH ACUTE MYELOID LEUKEMIA
Authors: Lind Evan F , Lamble Adam , Kosaka Yoko 
Published: January 2023

Abstract

Introduction
Acute myeloid leukemia (AML) is the most common blood cancer in adults. Despite intensification of chemotherapeutic regimens, survival rates have plateaued. AML is therefore a candidate for novel immune therapeutics such as immune checkpoint inhibitors (ICIs). We aim to identify the relevant lymphoid and myeloid cell subsets present in the bone marrow of patients with AML, evaluating their differentiation and activation status. We are also performing functional assays to further define the status of T cells in the AML microenvironment.

Methods
Immunophenotyping was performed on mononuclear cells from fresh AML patient bone marrow using mass cytometry consisting of 5 panels of 35 antibodies each. Functional assays measured both the proliferative capacity and cytokine production profile of bone marrow T cells in the presence of various ICIs.

Results
Bone marrow samples from 17 patients with AML and 7 healthy donors have been collected and analyzed to date.

Conclusion
Our results show that the immune microenvironment is very heterogeneous between patients, with some differential responses to ICIs suggesting that certain subsets of AML may be potential candidates for immunotherapy.
CD49a and CD103 expression on tumor infiltrating lymphocytes in B16-OVA tumors depends on location and is correlated with a naive phenotype
Authors: Melssen Marit Martina , Rodriguez Anthony B. , Slingluff Craig L. , Engelhard Victor H 
Published: January 2023

Abstract
Tissue resident memory (TRM) cells are found in certain peripheral tissues long after resolution of infections that generate them. TRM formation is induced by TGFβ and has been associated with upregulation of CD49a and CD103. Interaction of CD49a and CD103 with their respective ligands is thought to allow long term persistence of TRM cells in peripheral tissues. In melanoma patients, increased expression of CD49a and CD103 has been observed on tumor infiltrating lymphocytes (TIL) compared to PBMCs. These phenotypic similarities suggest that a subset of TIL might function as TRM. However, the impact of continuous antigen presentation and/or differential TGFβ expression levels on this property of TIL is currently unknown. Here we demonstrate that, in B16-OVA melanoma tumors, CD49a and CD103 expression on CD8 TILs is upregulated compared to lymph node (LN) derived CD8 T cells from non-tumor bearing mice. Additionally, relative expression of CD49a and CD103 differs between subcutaneous and intraperitoneal tumors. The capacity of T cells to persist in tumors thus potentially changes based on tumor location, which may be dependent on TGFβ levels. In contrast to LN of non-tumor bearing mice, CD103 is expressed on 50–90% of LN CD8 T cells in tumor bearing mice. Furthermore, CD103+CD8 TILs have a more naïve phenotype, whereas CD103neg CD8 TILs mainly express memory/effector T cell markers. The opposite is observed when T cells are activated in vitro. These data suggest that CD103+CD8 T cells in tumors are largely due to a tumor induced systemic induction of CD103 on naïve T cells, rather than intratumoral TRM differentiation. The importance of CD103 in retaining naïve T cells for functional differentiation within the tumor remains to be established.
Hypofractionated radiotherapy treatment preserves immune function and improves tumor control vs. hyperfractionated treatment.
Authors: Barlow Margaret Lauren , Battaglia Nicholas , Gerber Scott A , Lord Edith M 
Published: January 2023

Abstract
Radiotherapy is one of the most effective means of treating solid tumors in cancer patients. The radiation acts in two ways to control tumor growth. First, by inducing cell death due to direct damage to the DNA, then by the dying tumor cells releasing cellular signals that boost antitumor immunity. Unfortunately, radiotherapy may also damage the healthy tissue surrounding the tumor, causing adverse side effects in patients. To limit this damage, radiotherapy in the clinic is generally delivered as multiple small doses (hyperfractionation) over the course of several weeks. However, in preclinical radiotherapy studies, radiation is often given as a single or a few high ablative doses (hypofractionation). We previously observed improved tumor growth control and an increased number of tumor infiltrating lymphocytes (TIL) in tumors given an ablative dose when compared to a hyperfractionated scheme. As TILs are radiosensitive, we hypothesize that the multiple doses of a hyperfractionated scheme may result in increased killing of the TILs, reducing the effectiveness of the anti-tumor immune response. To further investigate this, we compared the responses of the mouse Colon38 tumor model to a hypofractionated vs. a hyperfractionated scheme, using doses based on current clinical protocols. In vitro, the hypofractionation treatment was more effective at killing tumor cells and inducing the tumor cells to produce IL-1β, TNFα, IL-6 and IP-10. In vivo, the hypofractionated treatment also resulted in a significant delay in tumor outgrowth and improved survival rates when compared to the hyperfractionated treatment. Therefore, hypofractionated radiotherapy may increase immunity and be more beneficial in tumor control. Funded by NIH R01CA28332.
miR-30e* is overexpressed in prostate cancer and promotes NF-κB-mediated proliferation and chemotherapeutic resistance
Authors: Egan Shawn M , Gollnick Sandra O 
Published: January 2023

Abstract
Prostate cancer (CaP) is a critical problem for the aging male population. CaP is managed using a combination of prostatectomy, androgen ablation, chemotherapy and radiation. Subsets of patients develop androgen-insensitive metastatic CaP for which therapeutic options for are limited. Androgen-insensitive CaP clinically presents with constitutive NF-κB activation. NF-κB activation in the primary tumor is correlated with metastatic disease, biochemical relapse and drives tumor promoting chronic-inflammation. The NF-κB pathway is tightly regulated by many highly conserved mechanisms, the most well characterized being IκBα which sequesters NF-κB in the cytoplasm. The mechanisms that perpetuate constitutive NF-κB activation in CaP are largely unknown. microRNAs (miRs) translationally repress a number of genes involved with NF-κB activation. We report that miR-30e* is significantly overexpressed in multiple murine CaP models. miR-30e* functions to repress the translation of IκBα and drive NF-κB activation. The miR:30e* : IκBα axis drives CaP cell proliferation. miR:30e* drives cyclin D1 and subsequent phosphorylation of Rb which facilitates CaP cell proliferation. Inhibiting the miR:30e* : IκBα axis is sufficient to significantly increase the survival of mice bearing prostate tumors and reduce tumor burden. Additionally, inhibition of miR-30e* sensitizes CaP cells to the chemotherapeutic docetaxel. Elevated miR-30e* is a mechanism that CaP cells utilize to bypass critical NF-κB regulatory checkpoints and hijack this pathway. Targeting the miR-30e* : IκBα axis provides a novel opportunity to repair a critical checkpoint to NF-κB activation and open a window of therapeutic intervention.
Adoptive transfer of induced pluripotent stem cell-derived regenerated T cells mediates effective regression of large tumors and establishes immunological memory
Authors: Ito Fumito , Saito Hidehito 
Published: January 2023

Abstract
Current approaches to adoptive T cell therapy are limited by the difficulty of obtaining sufficient numbers of T cells against targeted antigens with effective in vivo characteristics. Theoretically, this limitation can be overcome by using induced pluripotent stem cells (iPSCs) that could provide an unlimited source of autologous T cells. However, therapeutic efficacy of iPSC-derived regenerated T cells remains to be elucidated. To this end, we established iPSCs from Pmel-1 T cell receptor (TCR) transgenic CD8+ T cells specific for the gp100 melanocyte differentiation antigen as a preclinical model of evaluating antitumor reactivity of regenerated antigen-specific T cells from iPSCs (Pmel-1 iPSCs). Pmel-1 iPSCs retained the same rearranged configuration as the original Pmel-1 CD8+ T cells, and differentiated into CD4+CD8+ double positive (DP) T cells that expressed CD3 and the desired TCR in vitro. Stimulation of iPSC-derived DP T cells with the cognate antigen in the presence of IL-7 and IL-15 followed by expansion with IL-2, IL-7 and IL-15 generated large numbers of less-differentiated CD8+ single positive T cells with antigen-specific potent cytokine production and cytolytic capacity. Furthermore, adoptively transferred iPSC-derived CD8+ T cells escaped immune rejection, mediated effective regression of large tumors, improved survival, and established antigen-specific immunological memory. These findings support the translational potential of iPSCs to provide an unlimited number of phenotypically defined, functional and expandable autologous antigen-specific T cells harboring characteristics necessary for in vivo effectiveness.
Imprime PGG, a yeast β-glucan PAMP elicits a coordinated immune response in combination with anti-PD1 antibody
Authors: Qiu Xiaohong , Chan Anissa S.H. , Jonas Adria Bykowski , Kangas Takashi , Ottoson Nadine R. , Graff Jeremy R. , Bose Nandita 
Published: January 2023

Abstract
Imprime PGG (Imprime), a b glucan PAMP is currently in clinical development in combination with tumor-targeting antibodies, anti-angiogenic antibodies and immune checkpoint inhibitors. Randomized Phase 2 clinical trials of Imprime in the 1st-line treatment of stage IV non-small cell lung cancer have shown promising efficacy in terms of both objective tumor response and survival. As a PAMP, Imprime is readily recognized by innate immune cells, triggering immune responses including repolarization of M2 macrophages and dendritic cell (DC) maturation. Earlier results have shown that both the M2 macrophages and DC derived from Imprime-treated monocytes have higher expression of PD-L1 and CD86. Since PD-L1 expression, a marker for adaptive immune resistance, and CD86, a co-stimulatory marker for T cell activation could potentiate the activity of anti-PD1 antibody, we hypothesized that the combination will show synergistic T cell stimulatory activity. Ex- vivo treatment of T cells with anti-PD1 antibody, nivolumab showed increased proliferation in response to CD3/CD28 stimulation. Coculture of these T cells with Imprime-treated M2s or DCs drove further T cell expansion and increased production of several cytokines including IFN-γ, IL-2, TNF-α, GM-CSF, and IP-10. We next evaluated the anti-tumor efficacy of Imprime and murine anti-PD1 in a syngeneic tumor model. In the CT-26 syngeneic model, the median tumor volume in the anti-PD1 alone treatment group was 936 mm3, while it was only 172 mm3 in the Imprime and anti-PD1 combination group. Collectively, these data suggest that the combination of Imprime and checkpoint inhibitors holds a lot of promise for cancer immunotherapy.
Shortened T cell culture with IL-7 and IL-15 provides the most potent chimeric antigen receptor (CAR)-modified T cells for adoptive immunotherapy.
Authors: Ghassemi Saba , Bedoya Felipe , Nunez-Cruz Selene , June Carl , Melenhorst Jos , Milone Michael 
Published: January 2023

Abstract
Adoptive T cell immunotherapy involves the isolation, ex vivo expansion and reinfusion of T cells, and is dependent on T cell engraftment and persistence for efficacy. IL-2 is the most commonly used cytokine for ex vivo T cell culture; however, there is renewed interest in IL-7 and IL-15 due to their ability to enhance the survival and proliferation of stem cell memory (Tscm) T cells. Peripheral blood (PB) contains substantial numbers of Tscm, central memory (Tcm) and effector subsets. During ex vivo culture, PB T cells differentiate into cells with a predominantly CD45RO+, CD27−, CCR7− effector phenotype. Exogenous IL-7/15 delays this transition in phenotype and preserves a greater proportion of Tscm and Tcm cells. We hypothesize that limited ex vivo T cell culture in the presence of IL-7/15 rather than IL-2 will enhance engraftment and persistence of T cells in vivo. We show that T cells can be harvested from ex vivo cultures as early as day 3 (d3) following activation. T cells expressing a chimeric antigen receptor targeting CD19 (CART-19) show potent yet specific cytotoxicity in vitro. We investigated the therapeutic potential of cells harvested at d3 versus later time points using a Nalm-6 leukemic cell xenograft mouse model. We demonstrate that d3 CART-19 cells show potent anti-leukemic activity compared to day 5 or day 9 cells. Comparing CART19 cells cultured in IL-2 or IL-7/15 for 3 or 9 days, we show that mice treated with d3 cells cultured in IL-7/15 exhibit the greatest anti-leukemic efficacy at a 10-fold lower dose compared with day 9 cells. In summary, we show that limiting T cell culture ex vivo to the minimum required for lentiviral transduction, in the presence of IL-7/15, provides the most efficacious T cells for adoptive immunotherapy.