bluuebunny/crossref_metadata_2025_split huggingface dataset:
14-3-3zeta Regulates Transmembrane Receptor Signaling by Binding TRAF Proteins
Authors: Ayyasamy Rathinam , Chattopadhyay Saurabh , Chakravarti Ritu 
Published: August 2023

Abstract
Adaptor proteins are critical for signal transduction events involving cytokine signaling, host-response, and autoimmune diseases. Our laboratory is interested in understanding the immune regulatory role of 14-3-3zeta (z), an adaptor protein, in signal transductions and diseases. We examined the role of 14-3-3z in interleukin-17 (IL-17A) signaling and autoimmune inflammatory arthritis using genetically modified cells and rats. The IL-17A signal transduction requires IL-17 receptor cytoplasmic domain interactions with Act1 and several TRAF proteins. Our results show that 14-3-3z interacts with TRAF6, TRAF2, and TRAF5 upon IL-17A stimulation which has functional consequences. The interaction of 14-3-3z and TRAF protein was found to be specific, as mutation of the interacting residues resulted in the loss of binding with functional impacts. Further interaction studies show a higher affinity of 14-3-3z for TRAF6, which has two putative 14-3-3 binding sites. To examine the generality of 14-3-3z-TRAF interactions and their role in another transmembrane receptor signaling, we examined RANKL signal transduction. Similar to IL-17A, 14-3-3z-TRAF interaction was increased upon RANKL treatment and had functional consequences. While there are common domains and significant sequence conservation between the TRAF proteins, a specificity of interactions exists with 14-3-3z. Our results show that the 14-3-3z-TRAF axis is a novel regulatory mechanism underneath several transmembrane receptor-mediated signaling. Current studies target understanding the molecular basis of interactions between 14-3-3z and specific TRAF proteins.
Development of a specific, active human IL-18 assay for assessing inflammasome activation
Authors: Mamott Daniel B , Mahan Sarah , Cali Jim , Lazar Dan , O’Brien Martha 
Published: August 2023

Abstract
Interleukin-18 (IL-18) was first identified for its ability to induce interferon-γ expression. IL-18 is a proinflammatory cytokine and its release along with IL-1β is a primary output of inflammasome activation. We have developed convenient tools for monitoring inflammasome activation, including assays for caspase-1 activity and IL-1β release. While both IL-1β and IL-18 are expressed as inactive zymogens requiring processing to active forms, they differ in expression and regulation. IL-1β requires upregulation for expression and is primarily restricted to myeloid cells, whereas IL-18 is constitutively expressed more broadly in epithelial cells, as well as myeloid cells. IL-18 is also uniquely regulated by an IL-18 binding protein (IL-18bp). The IL-18bp has a very high affinity for active IL-18 and effectively blocks receptor activation. IL-18-stimulated interferon-γ release stimulates IL-18bp release in a negative feedback loop. The Lumit™ technology is a Nanoluc®-based platform that enables direct, no wash assays for analyte detection using standard luminometers. For Lumit IL-18 assays, we either NanoBiT®-labeled two IL-18 antibodies or NanoBiT®-labeled the IL-18bp as well as one IL-18 antibody. These resulted in, respectively, one assay that measures both IL-18 and proIL-18 and another assay specific for active IL-18. The specific assay was used to measure active IL-18 release from canonical inflammasome-activated THP-1 cells and PBMCs, as well as non-canonical inflammasome-activated lung A549 epithelial cells. Enabling direct quantitation of unbound, active IL-18 is essential for elucidating IL-18’s role in inflammation and disease. A Lumit™ IL-18 assay adds to the toolbox for monitoring inflammasome activation.
TRAF3 regulates IL-6 receptor signaling in T cells through the phosphatase PTPN22
Authors: Hornick Emma , Shao Peng , Butler Noah , Bishop Gail A 
Published: August 2023

Abstract
IL-6 is an important signal three cytokine for T helper 17 (Th17) and T follicular helper (Tfh) differentiation. Previous work has suggested a possible role for TRAF3 in the function and/or differentiation of these two subsets, but no direct studies have yet been performed. In this study, we examine the role of TRAF3 in IL-6R signaling in T cells. TRAF3 regulates IL-2R and type I interferon receptor signaling in T cells by altering recruitment of the phosphatases PTPN2 and PTPN22, respectively. B cell TRAF3 curtails IL-6R signaling by recruiting PTPN22, thus we hypothesized that TRAF3 regulates T cell IL-6R signaling through a PTPN22-dependent mechanism.
We investigated the role of T cell TRAF3 using a TRAF3-deficient human T cell lymphoma line and a T cells from a T cell-specific TRAF3 knockout mouse (T-Traf3−/−). We found reduced IL-6-induced STAT1 and STAT3 activation in human and mouse T cells. STAT1 and STAT3 activation are both required for full differentiation of mouse Tfh cells, whereas STAT3 is required for and STAT1 antagonizes Th17 differentiation. Interestingly, in vitro differentiation of naïve CD4 T cells to Th17 cells was not affected by TRAF3 deficiency. To determine the mechanism by which TRAF3 promotes STAT1 and STAT3 activation in response to IL-6R signaling, we treated WT and Traf3−/− T cells with IL-6 in the presence of PTPN22 inhibitor or vehicle control. We found that PTPN22 inhibition restored activation of STAT3, but did not restore activation of STAT1, suggesting different mechanisms of regulation. Together these data implicate TRAF3 in regulation of IL-6R signaling in a partly PTPN22-dependent manner, lending an additional level of understanding to TRAF3 regulation of Th17 and Tfh differentiation.
Supported by NIH R01 AI123107, R01 AI162656, T32 AI007260
B-cell activation state determines aryl hydrocarbon receptor regulation of CCR9 expression
Authors: Bauerle Logan V , Dekrey Gregory K 
Published: August 2023

Abstract
The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that regulates CCR9 expression on activated mouse B cells. Because AhR expression levels are themselves regulated by B cell activation state, in this study, we hypothesized that CCR9 regulation by the AhR would vary over time after B cell activation. This was tested in vitro using B cells isolated from mouse spleens (magnetic separation) followed by stimulation with 25 μg/mL LPS, 300 ng/mL anti IgD-dextran, and 2 ng/mL human TGF-Beta for 96 hours. Cells were exposed to AhR ligands at zero or 24 hours of culture as follows: 100 μM of the antagonist CH22319 or 50 nM of the agonist 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). CCR9 expression was determined by flow cytometry. As was shown previously, AhR agonism at zero hours of culture significantly enhanced CCR9 expression, relative to AhR antagonism (P &lt; 0.05). In contrast, AhR agonism at 24 hours of culture did not lead to significant alteration of CCR9 expression, relative to AhR antagonism. This finding suggests that migration of B cells to the gut may be enhanced by AhR activation, but only if AhR activation occurs simultaneously with antigen-mediated cell activation.
Supported by internal grants from the University of Northern Colorado, College of Natural and Health Sciences (NHS Student Research Fund, SBS Summer Research Fund).
Attenuated Salmonella blocks cancer metastasis by activating NK cells in an IFN-γ-dependent manner
Authors: Huang Jiandong , Lin Qiubin , Rong Li , Lin Shu-hai , Fu Guo 
Published: August 2023

Abstract
Metastasis accounts for 90% of cancer related deaths and blocking of metastatic cascade has critical clinical impact. However, the clinical drug development for cancer treatment, including cancer immunotherapies, is evaluated largely depending on their ability to cause tumour shrinkage and ignores the effect on metastasis as it has proven challenging to target. Therefore, there is an urgent need for novel therapeutic strategies and agents targeting metastasis. Using an attenuated Salmonella typhimurium strain YB1 engineered by our lab, we have found a potent suppressive effect of attenuated salmonella on cancer metastasis, regardless of cancer types and genetic background, by evoking strong anti-metastatic immune response. Further study showed that suppression of cancer metastasis by attenuated Salmonella only requires the innate immune response. Among the many induced cytokines, we identified IFN-γ as an indispensable factor for inhibiting cancer metastasis. Based on CyTOF (mass cytometry or cytometry by time of flight) analysis of the innate immune responses after Salmonella treatment and antibody-mediated cell depletion, we further demonstrated that NK cells are the major factor involved in Salmonella-provoked metastasis suppression. We found that IFN-γ was mainly produced by NK cells during early Salmonella infection, and in turn, IFN-γ promoted the accumulation, activation and cytotoxicity of NK cells. The IFN-γ-dependent NK cells directly eliminated newly accumulated cancer cells in the lung to block the cancer metastasis cascade in response to the Salmonella treatment.
This work was supported by Shenzhen Science and Technology Innovation Committee Basic Science Research Grant (JCYJ20170413154523577) to Dr. Jian-Dong Huang.
Targeting Fibrosis to Improve Cancer Outcomes
Authors: Breckenridge Joey Emery , Dell Garrison , Steele Hope , Cheng Jason Liang , Ghastine Andrew , Culberson Erica , Herro Rana 
Published: August 2023

Abstract
Scleroderma (SSc) is an idiopathic autoimmune connective tissue disease. The three hallmarks of SSc include 1) vasculopathy resulting in 2) progressive fibrosis or thickening of the skin, lungs, and internal organs and 3) aberrant immune infiltration. Fibrotic pulmonary involvement is the leading cause of death in SSc patients. Recent data from John Hopkins Scleroderma Center have shown a significantly increased risk of lung cancer in SSc patients. However, the mechanisms linking the two diseases are not well understood. Moreover, due to the close temporal relationship between SSc and lung cancer diagnosis, it is unclear whether lung fibrosis precedes tumorigenesis or conversely lung tumorigenesis precedes fibrosis. It has become evident that certain fibrotic diseases can progress towards cancer. Seemingly, non-alcoholic steatohepatitis or HBV/HCV infection-induced liver cirrhosis can progress to hepatocellular carcinoma. Vice versa, many solid tumors are fibrotic, and desmoplasia is referred to the pathological remodeling observed in the tumor microenvironment. There seems to be a tight link between fibrosis and cancer although the directionality of this progression is unknown. Moreover, several fibrotic processes involving the ß-catenin/Wnt, YAP/TAZ and hedgehog pathways in addition to fibrogenic growth factors (TGFß, VEGF, FGF) have been known to drive cancer. We are developing a murine model to study the mechanisms driving fibrosis to cancer progression in the lungs.
Proteolysis-targeting chimera against NR4A1 for cancer immunotherapy
Authors: Wang Lei , Xiao Yufeng , Master Rohan , Luo Yuewan , Zheng Guangrong , Zhou Daohong , Zhang Weizhou 
Published: August 2023

Abstract
Modulating tumor-infiltrating immune cells is the key mission for the development of cancer immunotherapy. Based on recent literature and our bioinformatic analysis, nuclear receptor subfamily 4 group A member 1 (NR4A1) is an ideal target for cancer immunotherapy due to its important role in T cell exhaustion, regulatory T (Treg) cell function. Here we targets NR4A1 via proteolysis-targeting chimeric (PROTAC) technology, expecting to achieve immune activation and cancer clearance.
We designed and synthesized over 80 NR4A1 PROTACs based on several reported NR4A1 ligands, such as celastrol. We found that several celastrol-based PROTACs achieved effective degradation of target protein NR4A1. We studied the therapeutic efficiency of the lead NR-V04, a celastrol- and von Hippel-Lindau (VHL)-based PROTAC. As expected, we confirmed that the NR-V04-induced NR4A1 degradation is through the uniquitin proteasome system with a tenery complex formation among NR4A1, VHL and NR-V04. The lead NR-V04 exhibits outstanding pharmacokinetics and effectively inhibits tumor growth at a low dose, 1.8mg/kg twice a week, in B16F10, Yummer 1.7, and MC38 mouse tumor models. We proved that NR-V04 significantly enhances anti-tumor immunity with seemingly distinct mechanisms of immune activation in a tumor-dependent manner. For example, NR-V04 induces the expansion of B cells and the reduction in myeloid-derived suppressor cells (MDSC) in B16F10; the reduction of Tregs, and the induction of CD8 effector population in Yummer 1.7 and MC38 tumors. we did not find obvious toxicity at these effective doses for up to 2 months. Therefore, the lead NR-V04 or other better ones under test have the potential for translation as a novel immunotherapeutic strategy.
BC200100 - “Developing a Novel PROTAC-Based NR4A1 Degrader for Breast Cancer Therapy” (DEPARTMENT OF THE ARMY)
YAP inhibition by CO promotes ferroptosis via ATF4-dependent REDD1 increase in melanoma
Authors: Park Jeongmin , Gong Jeong Heon , Joe Yeonsoo , Chung Hun Taeg 
Published: August 2023

Abstract
Ferroptotic cell death is a recently recognized form of non-apoptotic regulated cell death and exerts cytotoxicity in cancer cells. Carbon monoxide (CO) as an endogenous physiological mediator confers therapeutic potential in various cancer, while the therapeutic effects of CO on melanoma and its mechanism remain elusive. In this study, we suggest that CO induces ferroptosis via YAP inhibition and ATF4 activation in melanoma. YAP phosphorylation on serine 127 by CO-releasing molecule 2 (CORM2) caused cytoplasmic retention of YAP in B16F10 mouse melanoma cells, resulting in inhibition of YAP activity for transcription of target genes. Thus, CO-mediated YAP inhibition decreased SLC7A11 expression in melanoma cell lines, leading to ferroptosis. Moreover, CO increased REDD1 expression through PERK-ROS-ATF4 pathway. ATF4-REDD expression increased by CO became vulnerable to ferroptosis in melanoma. Thus, CO-induced YAP inhibition can induce ferroptosis via ATF4-mediated REDD1 increase and inhibition of SLC7A11. Furthermore, CO gas inhalation significantly inhibited B16F10 tumor size in xenograft mouse models resulting from ferroptosis induction via down-regulation of SLC7A11 and up-regulation of REDD1. These results suggest that CO acts as a potent inducer of ferroptosis in melanoma and that CO critically depends on YAP inhibition and ATF4 activation to promote ferroptosis.
Advancing Immunovirotherapy with Zika virus to treat high-grade brain tumors
Authors: Nair Sharmila , Chheda Milan , Diamond Michael S 
Published: August 2023

Abstract
High-grade brain tumors such as glioblastoma multiforme (GBM) are the most aggressive primary brain tumor and virtually all patients die within 2 years of diagnosis. Poor patient outcomes are caused by two known factors. First, presence of GBM stem cells (GSCs) which are a subpopulation of tumor cells that are resistant to radiation treatment and chemotherapy. GSCs drive tumor initiation and are the primary cause for tumor relapses. Currently, no treatment consistently kills these highly resistant cells. Second, the cold tumor microenvironment is filled with anti-inflammatory T cells and myeloid suppressor cells contributing to a weak anti-tumor immunological response. Oncolytic viral treatments are considered an attractive treatment option for high-grade solid tumors as they are engineered to replicate and destroy transformed cells as well as improve inflammatory immune responses against the tumor. However, none are specifically designed to kill GSCs. We previously tested and demonstrated that the natural tropism of Zika virus (ZIKV) can be harnessed to target and kill GSCs.
In vivo we demonstrated that intratumoral ZIKV treatment was enough to improve the survival outcomes of glioma bearing mice up to 40–60%. Importantly the oncolytic action of ZIKV extended beyond its anti-GSC effects. Reduction in tumor size and improved survival outcomes following oncolytic ZIKV treatment was caused by Perforin 1 activity in CD8+ T cells which clears tumor components not directly killed by ZIKV. In preliminary studies we observed that DC-2 and CCR2+ antigen presenting cell (APC) subsets are utilized by ZIKV to drive CD8+ T cells against GBM. This highlights the immunological barriers of GBM that can be overcome with ZIKV immunovirotherapy.
β-catenin inhibition using XAV939 nanoparticle promotes immunogenic cell death and suppresses tumor progression
Authors: Antony Ferrin , Kang Xeujia , Chen Pengyu , Babu Jayachandra , Suryawanshi Amol 
Published: August 2023

Abstract
Although the incidence of conjunctival melanoma is rare, it is a highly metastatic tumor with high recurrence and mortality rates. The current conjunctival melanoma treatment strategies, including surgical resection combined with chemo- or cryotherapy, have limited usefulness because of systemic side effects, recurrence, and metastases. Tumor-intrinsic Wnt/β-catenin pathway mediates immune exclusion in the tumor microenvironment (TME) and promotes tumorigenesis. Previous studies have shown that XAV939, a potent tankyrase inhibitor, can be used for the pharmacological inhibition of β-catenin. In this study, using tocopheryl polyethylene glycol 1000 succinate (TPGS)-based nanoparticle formulation for XAV939, we show that β-catenin inhibition suppresses melanoma cell proliferation and migration. Additionally, XAV939 nanoparticles promoted immunogenic cell death (ICD) of tumor cells with a significant extracellular release of ICD molecules, including HMGB1, calreticulin, and ATP. Further, using a mouse model of conjunctival melanoma, we show that β-catenin inhibition significantly suppresses conjunctival melanoma progression. Collectively, the local delivery of β-catenin represents a novel and safe approach to promoting ICD and activating anti-tumor immunity for combinatorial immune checkpoint inhibition therapies against vision and life-threatening ocular melanomas.
This study was supported by research funding from the Intramural Grants Program, Auburn University Research Initiative in Cancer (AURIC), and start-up funds from the Department of Pathobiology, College of Veterinary Medicine, Auburn University.
Zinc mediates the interplay between Th1/Treg differentiation and regulates anti-tumor immunity
Authors: Sugandha N , Rizvi Zaigham Abbas , Dalal Rajdeep , Adhikari Neeta , Awasthi Amit 
Published: August 2023

Abstract
Dietary supplements play an important role in determining the health and immunity. Zn supplement has been used for the dietary chemoprevention of cancer. However, some of the studies say that Zn is important for cell proliferation and cells in the Zn deficient condition arrested in the S-phase of cell cycle. Dietary Zn deficiency is known to inhibit several types of animal tumors. Cancer cells have upregulated Zn importers and increased Zn levels and Zn deficiency induces cell death via apoptosis. As Zn supplementation is widely used a detailed study analysing the effects of Zn on tumor progression has become the necessity of this hour.
In this study we have identified that providing higher doses of Zn supplement for longer duration during tumor development results in increased tumor burden. Zn supplementation during tumor development alters mainly CD4 T cell subtypes by suppressing T helper 1 (Th1) cell population and enhancing T regulatory (Treg) cell population. Zn supplementation significantly increases PD1 immune checkpoints on CD4 T cells, CD8 T cells and γδT cells. The degranulation marker CD107a or LAMP1 expression on NK cells and CD8 T cells decreased indicating the decrease in cytotoxicity among these cells by Zn supplementation.
Our findings open excellent therapeutic avenues for targeting cancer. Firstly use of Zn chelators as a tumor therapeutic agent. Zn chelators can be combined with α-PD1 mAb checkpoint therapy to overcome the drawbacks of immune checkpoint therapy and efficient treatment outcome. Secondly trace element analysis can become the forefront of cancer immunotherapy, as our study along with many other reports prove that the level of trace elements determines the tumor development and therapeutic outcome.
Supported by grants from Department of Bio-Technology (DBT), Ministry of Science and Technology, Govt. of India.
mRNA-HPV Therapeutic Vaccine Encoding E6 and E7 Improves Therapeutic Potential in HPV-mediated Cancer
Authors: Lee Seonghyun , Yoon Hyunho , Hong Seol Hee , Bang Eun-Kyung , Kwon Sung Pil , Kwak Hye Won , Park Hyo-Jung , Lee Jisun , Park Hyeong-Jun , Bae Seo-Hyeon , Youn Hyewon , Keum Gyochang , Lee Shee Eun , Nam Jae-Hwan 
Published: August 2023

Abstract
The viral E6 and E7 oncoproteins of human papillomavirus (HPV) are strongly associated with cervical cancer. We developed and assessed the efficacy of an mRNA-HPV therapeutic vaccine (mHTV), containing the E6 and E7 proteins, in C57BL/J mice. mHTV generated robust T cell-mediated immune responses and induced short-lived effector and E7 tetramer-specific T cells. mHTV immunization significantly suppressed tumor growth in mice implanted with TC-1 tumor cells and improved overall survival. The re-transplantation of tumor cells into mice that survived the primary tumor challenge after primary vaccination led to the complete inhibition of tumor progression in all mHTV-treated mice. This showed that mHTV can effectively suppress cancer recurrence. mHTV also increased the number of tumor-infiltrating lymphocytes in the tumor tissues. When mice were implanted with TC-1 tumor cells after mHTV vaccination, most of the immunized mice were protected from the tumor challenge. In conclusion, mHTV showed considerable potential as not only a therapeutic vaccine, by efficiently killing cancer cells, but also as a prophylactic vaccine by enhancing T cell-mediated immune responses.
The Brain Korea 21 Plus Program, Partially supported. The Ministry of Food and Drug Safety grant (grant number 22213MFDS421) The National Research Foundation of Korea (NRF) grant (grant number NRF-2021M3E5E3080558)
T-cell and B-cell receptor repertoire profiling for biomarker discovery
Authors: Chenchik Alex , Makhanov Mikhail , Liu Tianbing , Hu Dongfang , Diehl Paul , Kobzik Lester 
Published: August 2023

Abstract
T-cell receptor (TCR) and B-cell receptor (BCR) repertoire profiling, also referred to as adaptive immune receptor repertoire (AIRR) profiling, holds great potential for the understanding of disease mechanisms and for the development of new treatments in infectious disease, autoimmunity and immuno-oncology. This potential could be greatly improved by combining information about receptor clonotypes with immunophenotypes of T- and B-cells. We developed a new technology for combined profiling of all human TCR and BCR variable regions and phenotypic characterization of immune cells in the same workflow. The TCR and BCR immunophenotyping method involves RT-PCR amplification and sequencing of the CDR3 regions of the TCR and BCR genes, as well as determining the expression levels of the most informative T- and B-cell phenotyping genes. Results show that this method allows for comprehensive profiling of all seven TCR and BCR chains from a single sample, in a highly reproducible manner, directly from micro-samples including cancer tissue, whole blood, sorted cells and more. Bioinformatic analysis of the next-generation sequencing (NGS) data allows profiling of the TCR and BCR clonotypes and identifcation of major immune cell subtypes and their activation status. Preliminary data from rheumatoid arthritis blood samples and others will be presented.