bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cellulose hydrogels as delivery vehicle of immunomodulatory mesenchymal stem cells
Authors: Flores Andrea Sofia , Gonzalez Keishla , Domenech Maribella 
Published: June 2021

Mesenchymal stem cells (MSCs) are cells that have the capacity of differentiation and specialization in skeletal tissues, and more recently immunomodulatory properties. Upon stimulation MSCs secrete immunosuppressive factors that can benefit patients with inflammatory conditions. To prolong therapeutic benefit of MSCs following implantation, delivery strategies that provide protection from external damage are needed. In this study, we evaluated a cellulose gel as a vehicle for encapsulation and delivery of MSCs. Cellulose is biocompatible and non‐degradable in humans, and the use of hydrogels can control the release of anti‐inflammatory factors. To determine the effectiveness of using cellulose hydrogels as a delivery vehicle and establish the relationship between MSCs and cellulose, MSCs were cultivated into three‐dimensional spheroids in cellulose matrixes of 0.3% and 0.5%. Proliferation was measured by spheroid growth over the course of 10 days and was then tested for the secretion profile of its main immunomodulatory factor, indoleamine‐pyrrole 2,3‐dioxygenase (IDO), after stimulation with IFN‐γ +/−. These results were compared to two‐dimensional controls, where higher secretion of IDO was expected in the three‐dimensional models. Multiple spheroids were formed in each concentration instead of one large spheroid. Proliferation was directly proportional to cellulose concentration, although the difference in spheroid growth between both concentrations was considered insignificant. These results show that cellulose based materials can be used as a matrix for in‐vitro studies and as a delivery vehicle for immunomodulatory MSCs.Support or Funding InformationFinancial support from the National Institute of Health RISE program under grant number 1R25GM127191‐01This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Increased Endoplasmic Reticulum Amino Peptidase‐1 in Sickle Cell Disease: Role of Hypoxia
Authors: Echevarria‐Lorenzo Nanivette , Sanchez‐Real David , Irizarry Jose , Inostroza‐Nieves Yaritza , Rivera Alicia , Romero Jose 
Published: June 2021

Sickle cell disease (SCD) is a genetic disorder caused by a single amino acid substitution in the β‐globin gene that results in hemoglobin polymerization, deformed erythrocytes and endothelial cell activation leading to chronic inflammation, anemia, hypoxemia and vaso‐occlusive crisis (VOC). These events contribute to the development of acute and often severe and debilitating, painful episodes in patients with SCD through unclear mechanisms. In SCD, chronic hypoxia and subacute inflammation contribute to endothelial cell activation that is associated with increased inflammatory cytokine markers such as endothelin‐1 (ET‐1), tumor necrosis factor‐α (TNFα), and TNFα receptor‐1 (TNFR1). Endoplasmic Reticulum Aminopeptidase‐1 (ERAP1) is a ubiquitously expressed zinc‐metallopeptidase involved in regulating immune function and inflammation that cleaves cytokine cell surface receptors such as TNFR1 and plays an important role in the pathophysiology of ankylosing spondylitis. However, the relationship between ERAP1 and SCD is, to the best of our knowledge, unexplored. We first studied the effects of deoxygenation (5% O2) on ERAP1 mRNA expression levels by qRT‐PCR in the human endothelial cell line, EA.hy926. We observed a 3‐fold increase in the expression of ERAP1 after 4 hrs of hypoxia (p<0.05, n=4) and 4‐fold increase after 8 hrs of hypoxia (p<0.05, n=4); events that were associated with a 3.5‐fold increase in the hypoxia‐inducible factor 1‐α mRNA levels at these time points. Consistent with these results, activation of EA.hy926 cells with agonist such as angiotensin II was likewise associated with increased ERAP1 mRNA (p<0.05, n=4). We then studied two sickle transgenic knockout mouse models expressing human sickle hemoglobin, the βSAntilles and Berkeley (BERK) mice. We observed significant increases in ERAP1 mRNA levels (n=7 each; p<0.01) in kidneys from sickle transgenic mice when compared to control transgenic knockout mice expressing human hemoglobin A (HbA). In addition, and as previously documented in other sickle mouse models, our sickle mice had increased circulating TNFα by ELISA as compared to HbA mice (p<0.05, n=6–10 of each mouse type). In conclusion, increases in ERAP1 levels represent a novel target in SCD that is likely secondary to hypoxia and activated endothelial cells.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Modeling Melanoma in 3D uGel™‐based Artificial Tumor Tissues Demonstrate Initiation, Progression and Metastatic Processes In‐vitro
Authors: Lyden Timothy , Martin Mike 
Published: June 2021

In a continuing collaboration between our laboratory, the UWRF Tissue and Cellular Innovation Center, and BRTI Lifesciences, LLC we have been evaluating the application of a commercial hydrogel as a matrix for the development and study of several artificial tumor tissues. Here we report on the application of uGel™, a recently developed chitosan/hyaluronate precast matrix to generate 3D melanoma artificial tissues. In this work, we coupled uGel™ matrix with mouse melanoma cell lines B16F10 and B16F1 to generate and study 3D tumor tissues in‐vitro. As we have previously reported with another earlier and related hydrogel, Cell‐Mate™, both of these cells lines successfully produced extensive and long‐lived tumor tissues. In addition, in this series of studies with uGel™ it was possible to model not just the end stages of tumor progression and metastasis, but also the early stages of tumor establishment and local invasion of the matrix in patterns which clearly reflect early tumor development in vivo. As we had seen before, the final stages of tumor development in melanoma includes the generation of two distinct populations of cells, one pigmented and one not. Additionally these two populations also correlate tightly with differential patterns of tissue formation ranging from highly branched and pigmented to completely rounded, smooth and lacking virtually any pigment. Although the branching features are clearly associated with local invasion, our work suggests that the pale, rounded features may actually represent the metastatic stage for these tumors. Interestingly, uGel™ based melanoma developed and progressed to full‐blown tumor masses within a three week period while in our earlier work with Cell‐Mate™ constructs fully developed tumors required from 2–3 months of long‐term culture to reach the same stage. Although the two materials are biochemically identical, they do differ in mechanical properties at the time of loading which may play a role in the significant differences observed in terms of developmental timeline. This study further support our view that small‐scale and long‐term 3D artificial tissues are an effective tool to better understand the biology of cancer and to study it's progression in‐vitro.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Modeling Tumor Development, Progression and Metastasis in 3D Artificial Tissues: Characterization of Structure and Labeling Patterns in Breast Cancer Hanging‐drop Spheroid Cultures
Authors: Klinkhammer Katie , Leopold Marissa , Lyden Timothy 
Published: June 2021

In this report our laboratory presents the results of a series of studies being conducted to explore the application of 3D artificial tumor tissues as in‐vitro models of breast cancer development, progression and metastasis. In this case, we have employed MCF‐7 breast ductal adenocarcinoma cells and 3D Biomatrix hanging drop plates to evaluate these tumor properties within the context of staged tumor spheroids. A consistent observation of ours in nearly a decade of modeling breast cancer using various natural 3D scaffolds and hydrogel matrices has been that solid tumor models grown in long‐term culture predictably begin to shed both single cells and cell clusters in the form of small groupings of cells as well as progressively larger spheroids. Although we have shown that at least some of these structures have properties consistent with metastasis, including attachment and invasion of target tissues, spontaneously generated versions of cancer clusters also present as pleiomorphic structures and in relatively small numbers which complicates their study. Therefore, we sought to use a system that generates identical clusters and spheres from specific numbers of suspended single cells. The hanging drop system we employ comes in a 96 well format that allows us to generate literally as many spheres as we need for each study. Furthermore, these spheres can be grown out to match any of the sizes commonly found in our tumor models thereby allowing us to study each stage for structure, cell populations and markers. In addition these spheres are also assisting us in systematically studying the process of cancer cell cluster attachment and invasion and to test our working hypothesis that this is controlled through activation of EMT‐like responses through CD44 binding of hyaluronate on the target tissue. Using direct labeling of intact spheres coupled to embedding and sectioning followed by immunolabeling, these studies are demonstrating that over a period of 144 hours our spheroid cultures progress from simple clusters of a few cells to aggregated spheres containing thousands of cells and measuring up to ~600um in diameter. By this later stage, many spheres actually begin to generate their own smaller spheres and clusters as well. In our view this reflects the concept that past 48 hours in culture these spheres transition into being actual tumor organoids themselves. This is supported by development of a complex CD24+ internal structure distinct from an outer shell of CD44+ cells. These studies are ongoing and support our view that 3D artificial tumors are effective tools to study cancer biology in‐vitro.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Developing a method of co‐incubating murine adipocytes and macrophages for evaluating cellular communication
Authors: Beck Jordan , Caldari‐Torres Cristina 
Published: June 2021

Adipose tissue, an organ containing both adipocytes and adipose tissue macrophages (ATMs), secretes cytokines known as adipokines. Of these two cell types, ATMs have a larger role in the production and release of proinflammatory adipokines. The objective of this research was to develop a system of co‐incubating adipocytes and macrophages that allowed us to quantitatively measure the cross‐talk between these two cell types. Nitric Oxide (NO) and interleukin‐6 (IL‐6) concentrations were quantified to determine macrophage activation and production of a proinflammatory adipokine, respectively. Leptin concentrations were used as a measure of adipocyte‐specific adipokine production. The cell lines used were RAW 264.7 (murine macrophages) and 3T3‐L1 (murine fibroblasts). 3T3‐L1 fibroblasts were differentiated into adipocytes, which was confirmed by the presence of lipid droplets. Transwell plates with permeable inserts were used to co‐incubate the macrophages and adipocytes; the confluent adipocytes were maintained in the 12‐well plate (lower compartment) and the macrophages were added to the transwell insert (upper compartment). Macrophages were stimulated with 0.01 mg/ml of LPS either before or after the addition of the the macrophages to the insert. The treatments were: 3T3+media, 3T3+media+LPS, 3T3+unstimulatedRAW, 3T3+RAW+LPS, 3T3+stimulatedRAW (in fresh media), and 3T3+stimulatedRAW (in conditioned media). There was no difference in IL‐6 concentration measured in the upper compartment of the transwell insert vs the lower compartment, while measured NO levels were higher in the upper compartment and leptin levels were higher in the lower compartment (p<0.001, matched pairs t‐test). The 3T3+RAW+LPS, 3T3+stimulatedRAW (in fresh media) and 3T3+stimulatedRAW (in conditioned media) co‐incubation protocols resulted in the highest concentration of IL‐6 (p=0.0006). The 3T3+RAW+LPS and 3T3+stimulatedRAW (in conditioned media) co‐incubation methods yielded the highest concentrations of NO in both the upper and bottom compartments of the plate (p<0.001). There was no difference in leptin concentrations due to co‐incubation protocol. There was a positive relationship between NO and IL‐6 in both the top and bottom compartments (p<0.001, linear regression) and between NO and leptin in the top (p=0.03) but not in the bottom compartment. There was no significant relationship between IL‐6 and leptin concentrations in either the top or bottom compartments. For future experiments, our lab will utilize the co‐incubation protocols that resulted in the highest production of NO and IL‐6: 3T3+RAW+LPS and 3T3+stimulatedRAW (in conditioned media).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Differential Expression g‐H2AX and RAGE in the Placenta of Gestational Diabetes Mellitus (GDM), Preterm Labor (PTL) and Preeclampsia Patients
Authors: Tsai K YF , Hirschi K M , Davis T , Llavina S , Knowlton M N , Bennet A , Reynolds P R , Arroyo J A 
Published: June 2021

The integrity of the genome is fundamental for human health. Defective DNA repair response (DRR) is associated with several diseases, including pulmonary fibrosis, neurodegeneration, and in diseased placenta. The most destructive form of genomic damage is known to be DNA double strand breaks (DNA‐DSBs), in which both strands of DNA are ruptured. g‐H2AX is a component of the histone octomer and is phosphorylated in response to DNA‐DSBs. The receptor for advanced glycation end‐products (RAGE) is a multi‐ligand receptor primarily expressed on cell membranes, where it functions as a progression factor in inflammatory signaling; however, the nuclear isoform of RAGE (nRAGE) has been described as having a crucial role in DNA‐DSBs repair. The purpose of this study was to identify placental nRAGE and g‐H2AXexpression during normal gestation (Control), gestational diabetes mellitus (GDM), intrauterine growth restriction (IUGR), preterm delivery (PTL) and preeclampsia (PE). Immunohistochemistry was used to determine localization of both molecules present in human placenta. Immunoblot was further used to quantify nRAGE andg‐H2AX production. Assessing DNA degradation was used to confirm placental DNA damage during pregnancy complications. Relative to controls we observed: 1) Increased staining for of RAGE and g‐H2AX in PTL and PE placentas; 2) increased placental nRAGE protein in the PTL and PE placentas; 3) increased placental g‐H2AX protein in both PTL and PE conditions; and 4) increased DNA damage in the diseased placenta. These research suggest a potential role for nRAGE during DNA‐DSB detection and repair in human PTL and PE placentas. These results may provide insight into the physiological relevance of these molecules, and if so, their modification during gestation may help alleviate placental disease.Support or Funding InformationThis work was supported by a grant from the Flight Attendant's Medical Research Institute (FAMRI, PRR and JAA) and a BYU Mentoring Environment Grant (JAA and PRR).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Lysophosphatidylcholine promotes trained immunity of human aortic endothelial cells via histone 3 lysine 14 acetylation‐dependent and independent gene modulation
Authors: Sun Yu , Lu Yifan , Yang William , Yu Daohai , Saaoud Fatma , Shao Ying , Cueto Ramon , Fu Hangfei , Wang Hong , Yang Xiaofeng 
Published: June 2021

We previously reported that a conditional danger associated molecular pattern (conditional DAMP), pro‐atherogenic lipid lysophosphatidylcholine (lysoPC) induces human aortic endothelial cell (HAEC) activation via histone 3 lysine 14 acetylation (H3K14Ac); and HAEC activation plays important role in vascular inflammation and atherosclerotic cardiovascular diseases. However, the mechanisms underlying how lysoPC induced HAEC activation via histone acetylation remain unknown. To examine this issue, we carried out RNA‐Seq of lysoPC‐treated HAEC; and CHIP‐Seq using antibody to H3K14Ac in lysoPC‐treated HAEC. By analyzing the RNA‐Seq and CHIP‐Seq data, we found out the following results: 1) lysoPC modulates the expressions of 9 out of 31 histone acetyltransferases (HATs) with 6 increased and 3 decreased; and three out of the 6 increased HATs are reported to mediate H3K14Ac; 2) among three Acetyl‐CoA generating enzymes, lysoPC upregulates ACS2 (nucleus) and ACL (cytosol), and decreases ACS1 (peroxisome); 3) lysoPC modulates 5 out of 9 glycolytic enzymes with 3 increased and 2 decreased; and increases 6 out of 7 mevalonate pathway enzymes; 4) lysoPC increases the expressions of both T cell costimulation ligands and receptors; 5) lysoPC‐induced H3K14Ac increases the frequencies of downstream chromatin long range interactions (CLRIs); and lysoPC potentially‐induced other histone acetylations prefer to modulate upstream CLRIs. Our findings have demonstrated that lysoPC promotes HAEC activation and gene modulations via increasing acetylation of H3K14 and potential other histone lysines, and thereby frequencies of chromatin long‐range gene interactions, which significantly enhance newly characterized trained immunity features of HAEC. Our results provide novel therapeutic targets for controlling endothelial cell activation, vascular inflammation, and cardiovascular diseases.Support or Funding InformationThis work was supported by NIH grant to Drs. XF. Yang, H. Wang.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Elucidating the Expression and Role of Epithelial‐Derived Gasdermin‐B (GSDMB) in the Context of Chronic Intestinal Inflammation
Authors: Rana Nitish , Pizarro Theresa T 
Published: June 2021

Inflammatory bowel disease (IBD) represents a group of disorders that results in chronic, uncontrollable inflammation of the gastrointestinal (GI) tract. Despite recent advances, the precise etiology of the two major forms of IBD, Crohn's disease and ulcerative colitis, remains elusive. Gasdermin‐B (GSDMB), a novel molecule belonging to the Gasdermin family of proteins, has gained recent interest for its potential role in the pathogenesis and progression of chronic inflammatory disorders, particularly those that affect mucosal surfaces. Importantly, single nucleotide polymorphisms (SNPs) within the gene encoding for GSDMB have been reportedly associated with increasing the incidence of both asthma and IBD. Here, we aim to examine the aberrant expression and function of epithelial‐derived GSDMB towards the development and progression of chronic intestinal inflammation, such that observed in IBD. Using primary biopsy specimens derived from IBD patients, we confirmed the localization of GSDMB to primarily intestinal epithelial cells (IECs), both in protein and mRNA forms, via western‐blot techniques and immunohistochemistry staining. Moreover, preliminary evidence suggests an inflammasome dependent regulation of GSDMB (similar to that of the closely related GSDMD protein), via stimulation of IEC lines using lipopolysaccharides (LPS) and nigericin treatments to increase GSDMB mRNA expression. Due to the pore‐forming ability of the Gasdermin family of proteins, we suggest increased GSDMB expression may act as a membrane‐bound protein to facilitate the disruption of mucosal barrier function (i.e. restitution and repair), and act as conduit for release of inflammatory cytokines during the progression of IBD. Taken together, this data confirms an increased expression of epithelial‐derived GSDMB in the setting of chronic intestinal inflammation, providing indication for the role of GSDMB towards the development of inflammation within the gut mucosa during IBD.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Endothelium‐Specific Acid Ceramidase Gene Deletion Enhances Exosome Secretion and Release of NLRP3 Inflammasome Products during Hyperglycemia
Authors: Yuan Xinxu , Bhat Owais M. , Lohner Hannah , Li Pin‐Lan 
Published: June 2021

Exosomes have been demonstrated to be one of the mechanisms mediating the release of a variety of intracellular signaling molecules to mediate cell‐to‐cell communication. However, it remains unknown whether exosomes mediate the release of Nod‐like receptor pyrin domain 3 (NLRP3) inflammasome products such as IL‐1β from endothelial cells and how exosome‐mediated‐inflammatory signaling is regulated. The present study hypothesized that lysosomal acid ceramidase (AC) determines the fate of the multivesicular body (MVB) to control the exosome‐mediated release of NLRP3 inflammasome products during hyperglycemia. Using streptozotocin (STZ)‐induced mouse model of diabetes mellitus (DM), we found that endothelium‐specific AC gene knock‐out mice (Asah1fl/fl/ECcre) had significantly enhanced formation and activation of NLRP3 inflammasomes including IL‐1β production in the coronary arterial endothelium. These mice also had increased thickening of the coronary arterial wall and reduced expression of tight junction protein (ZO‐1 and ZO‐2) compared to their wild‐type (WT/WT) littermates. Interestingly, we observed that the expression of exosome markers such as CD63 and alkaline phosphatase (ALP) was also augmented in STZ‐treated Asah1fl/fl/ECcre mice compared to WT/WT mice, which was accompanied by an increased IL‐1β release of exosomes into the blood. In the primary cultures of coronary endothelial cells (CECs) from WT/WT and Asah1fl/fl/ECcre mice, we demonstrated that the AC deficiency markedly enhanced the formation and activation of the NLRP3 inflammasomes, but significantly down‐regulated tight junction proteins when CECs were exposed to high levels of glucose. The CECs from Asah1fl/fl/ECcre mice also had decreased MVBs and increased exosome release in response to high glucose stimulation and these exosomes contained increased IL‐1β. These results from our in vivo and in vitro experiments suggest that AC importantly controls exosome secretion and release of NLRP3 inflammasome products in CECs and its deficiency enhances this exosome‐mediated release of NLRP3 inflammasome products to activate coronary arterial inflammatory response during hyperglycemia.Support or Funding InformationHL057244, HL075316, and DK120491This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.