bluuebunny/crossref_metadata_2025_split huggingface dataset:
HE4 Promotes Events Associated with Metastatic Ovarian Cancer Via Regulation of the Extracellular Matrix
Authors: Gaudet Hilary , Ribeiro Jennifer , Khan Mehreen , Schorl Christoph , James Nicole , Oliver Matthew , Singh Rakesh , DiSilvestro Paul , Moore Richard , Yano Naohiro 
Published: June 2021

Human epididymis protein 4 (HE4) has diagnostic and prognostic abilities for epithelial ovarian cancer. The Risk of Malignancy Algorithm (ROMA) uses HE4 serum levels in combination with other factors to detect and monitor ovarian cancer. Several studies have demonstrated a role for HE4 in invasion and migration. Here, we further investigated the role of HE4 in events associated with metastatic ovarian cancer. Treatment of OVCAR8 ovarian cancer cells with recombinant HE4 (rHE4) promoted invasion, haptotaxis toward a fibronectin substrate, and adhesion onto fibronectin. In addition, overexpression of HE4 or treatment with rHE4 promoted the upregulation of transcripts coding for extracellular matrix proteins, including LAMC2 and LAMB3. Deposition of laminin‐332, the heterotrimer comprising LAMC2 and LAMB3 proteins, was increased in OVCAR8 cells treated with rHE4 or conditioned media from HE4‐overexpressing cells. Enzymatic activity of matriptase, a serine protease that cleaves laminin‐332 and contributes to its pro‐migratory functional activity, was enhanced by rHE4 treatment. Focal adhesion kinase signaling was activated in OVCAR8 cells treated with conditioned media from HE4‐overexpressing cells and focal adhesions were increased in cells treated with rHE4 in the presence of fibronectin. These results indicate a role for HE4 in mediating events associated with metastatic ovarian cancer.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
MicroRNA‐203 regulates cell junction and adhesion proteins by modulating Zeb1/Vimentin axis in breast cancer cells after PARP1 inhibition.
Authors: SHUKLA NIDHI , Rajawat Jyotika , Mishra Durga Prasad 
Published: June 2021

PARP inhibitors are currently in trend to treat various type of cancers and they have shown promising results in homologous recombination deficient breast and ovarian cancers like those bearing BRCA1/2 mutations. There are many reports describing about the failure of PARP inhibitors due to resistance or unknown factors in clinical trials. It is crucial to look into in‐depth molecular mechanisms behind the role of PARP in cancer pathogenesis and long term effect of PARP inhibition. MicroRNAs (miRNAs) are small non‐coding RNAs, acting as post‐transcriptional regulators of gene expression and known to regulate variety of cellular processes, including the DNA damage response and regulation of cell junction and adhesion proteins. However, little is known about miRNAs mediated regulation of sensitivity to PARP inhibitors and its pathological significance in context to regulation of cell junction and adhesion proteins. Aberrant miRNA regulation of junction proteins is one of the leading causes of cancer pathogenesis. Keeping these views in consideration, we aimed to identify novel miRNAs involved in the regulation of cell junction and adhesion proteins in breast cancer malignancy after PARP inhibition. To investigate this, we performed miRNA microarray analysis of PARP inhibitor treated MDA‐MB‐231 cells and found upregulation of tumor suppressor miR‐203 after PARP inhibition. PARP1 inhibition led to reduced expression of vimentin and Zeb1 while ZO‐1, E‐cadherin and RhoGDI were upregulated in breast cancer cells. To confirm the whether PARP inhibitory effects were mediated by miR‐203, cells were transfected with miR‐203 mimic and anti‐miR. We observed that miR‐203 mimic resulted in inhibition of junction proteins Zeb1 and vimentin, coinciding with PARP inhibition. We also found that PARP inhibition caused reduced cell growth and proliferation of breast cancer cells which correlated with the overexpression of miR‐203. Our findings indicates that upregulation of miR‐203 due to PARP inhibition is involved in regulating cell junction proteins vimentin and zeb1 as well as cytosketelal RhoGTPase and thus regulating cell migration, invasion and proliferation. Taken together, these observations raise the possibility that these miRNAs could be used as biomarkers to identify patients that may benefit from treatment with PARP inhibitors.Support or Funding InformationJR acknowledge the support from Department of Biotechnology (DBT) for DBT Bio‐CARe fellowship. NS acknowledges the support by the senior research fellowship from the Indian Council of Medical Research (ICMR). This work was supported by the grants from Department of Biotechnology (GAP0187) to DP Mishra.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inhibition of endothelial SCUBE2, a novel VEGFR2 co‐receptor, suppresses tumor angiogenesis
Authors: Lin Yuh‐Charn , Liu Chun‐Yu , Kannagi Reiji , Yang Ruey‐Bing 
Published: June 2021

ObjectiveSignal peptide‐CUB‐EGF domain containing protein 2 (SCUBE2), expressed on the endothelial cell (EC) surface, functions as a novel co‐receptor for vascular endothelial growth factor receptor 2 (VEGFR2) and enhances VEGF‐induced signaling in adult angiogenesis. However, whether SCUBE2 plays a role in pathological angiogenesis and whether anti‐SCUBE2 antibody is an effective strategy for blocking tumor angiogenesis remains unknown. The aim of this study was to investigate the pathological role and targeting therapy of SCUBE2 in tumor vasculature.Approach and ResultsImmunohistochemistry revealed that SCUBE2 is highly expressed in ECs of numerous carcinomas. Genetic EC knockout of SCUBE2 and pharmacological inhibition with the anti‐SCUBE2 monoclonal antibody (mAb) SP.B1 significantly reduced xenograft tumor growth, decreased tumor vascular density, increased apoptosis and decreased the proliferation of tumor cells. Mechanistic studies revealed that SP.B1 binds to SCUBE2 and induces its internalization for lysosomal degradation, thereby reducing its cell‐surface level and inhibiting the binding of and downstream signaling of VEGF, including VEGFR2 phosphorylation and AKT/mitogen‐activated protein kinase (MAPK) activation. Importantly, dual combination therapy with anti‐SCUBE2 mAb and anti‐VEGF antibody or chemotherapy was more effective than single‐agent therapy.ConclusionsEC‐surface SCUBE2 is a VEGFR2 co‐receptor essential for pathological tumor angiogenesis, and anti‐SCUBE2 mAb acting as an internalization inducer may provide a potent combination therapy for tumor angiogenesis.Support or Funding InformationThis work was supported by grants from Academia Sinica and the Ministry of Science and Technology (MOST) of Taiwan (VTA 106‐T‐6‐2, MOST 106‐2320‐B‐001‐017‐MY3, MOST 106‐0210‐01‐15‐02, MOST 105‐0210‐01‐13‐01, MOST 104‐0210‐01‐09‐02, MOST 104‐2320‐B‐001‐011‐MY3) to R.‐B.Y.
Defensin γ‐Thionin from Capsicum chinense Induces Apoptosis in the Human Breast Cancer Cell Line MCF‐7 and Regulate Histone H3 Epigenetic Modifications
Authors: Arceo‐Martinez Maria Teresa , Guzmán‐Rodríguez Jaquelina , Palomera‐Sánchez Zoraya , Ochoa‐Zarzosa Alejandra , López‐Meza Joel Edmundo 
Published: June 2021

Cellular homeostasis is regulated by genetic and epigenetic mechanisms controlling cell proliferation and differentiation; dysregulation in these functions has been associated to cancer. Breast cancer is considered the first worldwide cause of cancer death in women. Antimicrobial peptides (AMPs) have showed cytotoxic activity against diverse cancer cells through different mechanisms; however, only the effects of animal AMPs have been well characterized. Therefore, the aim of this work was to determine the cytotoxic effect of the plant defensin γ‐thionin on MCF‐7 breast cancer cells and to characterize its mechanism of action.The cytotoxic effect of γ‐thionin on MCF‐7 cells was assessed by trypan blue exclusion assays. This AMP was cytotoxic in a concentration‐dependent manner, with an IC50=117.29 μg/ml. Interestingly, viability of normal bovine mammary epithelial cells and peripheral blood mononuclear cells was unaffected. Additionally, this AMP induced apoptosis in MCF‐cells and did not affect the membrane integrity or cytosolic calcium. Further, γ‐thionin IC50 increased the expression of apoptosis related genes such as caspase 7, 8, 9, Apaf‐1 and cytochrome c. Transcriptional regulation has been linked to H3 epigenetic marks, for this reason, the global levels of H3 modifications were determined by western blotting. Data showed that γ‐thionin IC50 increases the mark of active transcription H3K9ac after 24 h of treatment. Intriguingly, the level of H3S10ph in cells treated with the defensin was higher at 12 h, which suggests an active proliferation mark. The mRNA expression of desacetylases HDAC1, HDAC2 and demethylase KDM4A was increased after 6 h of γ‐thionin treatment, while the expression of the kinase AURKB was induced at 12 h. Also, the expression of estrogen receptor gene (ESR1) was increased at 6 h and decreased at 24 h. Taken together, these results show that γ‐thionin induces apoptosis in MCF‐7 breast cancer cells and regulates changes in genetic expression, which could be associated with H3 epigenetic modifications. This is the first study of a plant AMP inducing cytotoxicity through epigenetic mechanisms in a hormone response breast cancer model.Support or Funding InformationCIC‐UMSNH 14.5This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Evidence of natural transcriptome regulation by cinnamon extract identified by changes in Akt1 mRNA levels of MCF‐7 breast cancer cells
Authors: Hill Matthew , Hall Seth , Almehmadi Seham , Lin Zachary , Aulthouse Amy , Kinder David , Stockert Amy 
Published: June 2021

Research in recent years has focused heavily on different tumorigenesis mechanisms. One such mechanism is the anti‐apoptotic effects regulated by varying Akt isoforms. Akt1 signaling can result in nuclear exclusion and subsequent cytosolic degradation of key transcription factors. It is proposed that transcriptional regulation of Akt1 offers potential benefits for both preventative and adjunct therapy for cancer treatment. Many natural products including cinnamon contain high amounts of polyphenolic compounds that have been shown to regulate both signaling proteins as well as key transcription factors. Previous research in the lab has suggested that cinnamon extract treatment of MCF7 breast cancer cells leads to epigenetic changes, specifically methylation patterns. Preliminary experiments demonstrate potential methylation changes at the Akt1 gene that alter expression. Real‐time PCR was used to evaluate the expression levels in MCF7 breast cancer cells under cinnamon treatment conditions and vehicle. Duplicate cell sets were grown to allow separate isolation of both DNA and RNA. Cells were grown to 70–80% confluence and treated with either cinnamon extract or vehicle for 24 hours. Cells were collected at the 24 hour time point by trypsin release and stored at −80 degrees C until use. Analysis of mRNA levels of Akt1 suggest a change in Akt1 expression level that is proposed to result from changes in methylation. In order to better substantiate the potential change in Akt1 methylation, MethylLight ® was completed with Akt1 specific primers and results were compared to the observed mRNA levels. Further research will allow comparison of Akt1 location under these conditions as well as Akt1 activity.Support or Funding InformationDepartments of Pharmaceutical and Biomedical Sciences and Biological and Allied Health Sciences, Ohio Northern UniversityThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Copper Binding Regulates Cellular Prion Protein Function
Authors: Nguyen Xuan T. A. , Tran Hoa T. , Cojoc Dan , Legname Giuseppe 
Published: June 2021

The cellular prion protein (PrPC), mainly known for its role in neurodegenerative diseases, is involved in several physiological processes including neuritogenesis. By combining genomic approaches, cellular assays and focal stimulation technique, we have explored the molecular mechanism underlining PrPC function as a signaling molecule in neuritogenesis. Several recombinant prion protein (recPrP) mutants were obtained to treat primary hippocampal cultures in bulk or exposed near the hippocampal growth cones (GC) of single neurons in a local stimulation manner. While focal stimulation of GC with wild‐type recPrP induced neurite outgrowth and rapid GC turning towards the source, N‐terminal mutants fail to support this function.In particular, the copper‐binding sites mutants present at the N‐terminus of PrPC are toxic to neurons indicating this region being crucial for the function of the protein. Mutants of recPrP including a key mutation for prion conversion H95Y (Giachin, Mai et al. 2015) or a GSS‐linked mutation abolish the function on neuritogenesis. Altogether, our findings indicate the functional regions for PrPC involved in neuritogenesis, suggest a potential link between loss‐of‐function of the protein and disease initiation.Support or Funding InformationFondo per gli Investimenti della Ricerca di Base (FIRB) program project (grant number RBAP11FRE9_001 to GL) from Ministero dell'Istruzione, dell'Università e della Ricerca(MIUR), ItalyThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Spectral Sensitivity of Sensory Motor Integration of Gill Lateral Cell Cilia in the Bivalve Mollusc Crassostrea virginica
Authors: Buchanan Reniece , Jean‐Pierre Johanne , Catapane Edward J. , Carroll Margaret A. 
Published: June 2021

Gill lateral cells of Crassostrea virginica are innervated by serotonin and dopamine nerves from their ganglia. Most bivalves have lateral cell cilia that respond to serotonin, which is cilio‐excitatory, and dopamine, which is cilio‐inhibitory. The motor aspects of gill lateral cell innervation have been well studied, but not the sensory side. Recently we found various sensory cues, including light, initiated a sensory‐motor integration response between mantle rim sensory tentacles and gill lateral cell cilia, via the visceral and cerebral ganglia. We also found that in animal preparations in which their shells removed, the cerebral ocelli (eyespots) responded to light causing lateral cell cilia to decrease beating rates. We hypothesize that light can penetrate oyster shell and stimulate cerebral ocelli to initiate a motor response of gill lateral cell cilia. In addition we hypothesize that if light does penetrate the shell, certain wavelengths will be more effective in stimulating sensory cells in ocelli and mantle rim tentacles. To test this we first measured light transmittance through shells using a Beckman DU800 spectrophotometer and found red light (650+ nm) produced the greatest degree of transmittance through the shell. Using animal preparations with intact gill to cerebral/visceral ganglia innervation we found that white light shown though the shell stimulated ocelli to initiate a motor response to slow down gill lateral cell cilia beating rates. Cilia beating was measured by stroboscopic microscopy and expressed as beats/sec ± sem. Then using different wavelengths of light (blue ‐ 405 nm, green ‐525 nm, amber ‐ 591 nm, red – 680 nm) we stimulated mantle rim sensory tentacles and cerebral ocelli with the shell in place. Under these conditions we found that only red light resulted in a sensory‐motor response that slowed lateral cell cilia beating rates. Cilia beating rates slowed from a basal rate of about 15 beats/sec to zero over a 20 minute period. These results were statistically significant and the other tested wavelengths had no significant effect on cilia beating rates. This study further demonstrates the integration of photosensory signals in the control of gill lateral cell cilia in the bivalve C. virginica, and adds new knowledge that demonstrates a spectral sensitivity of sensory cells in the cerebral ocelli and mantle rim tentacle involved with this sensory‐motor integration response.Support or Funding InformationSupported by grant 690340047 of PSC‐CUNY, grant 2R25GM06003 of the Bridge Program of NIGMS and a Carnegie Foundation award.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Gene Expression in the Rat Thalamus following Chronic Neuropathic Pain Development: mRNA sequencing analysis
Authors: Clifford John L. , Kumar Raina , Srinivasan Seshamalini , Dmitrov George , Gautam Aarti , Walsh Andrianna , Workman Emily , Chavez Roger , Sosanya Natasha , Christy Robert J. , Hammamieh Rasha 
Published: June 2021

Chronic pain afflicts roughly 44% of Service Members returning from Iraq and Afghanistan, and it is estimated that 100 million Americans suffer from this condition. The underlying molecular mechanisms of chronic pain development have remained elusive. The broad objective of this study is to identify molecular signals and indications of chronic pain in key brain regions during the onset and progression of neuropathic pain, in a rodent model. For this experiment, we collected tissue from five brain regions (thalamus, amygdala, cingulate cortex, insular cortex, and somatosensory cortex), as well as the L4–L6 spinal cord and L5 dorsal root ganglia, from rats subjected to the spared nerve injury (SNI) protocol, a model for chronic neuropathic pain. Tissue was collected at 1, 3, 7, 14 and 21 days post‐injury from each tissue and RNA extracted for mRNA sequencing (TruSeq total RNA protocol and Illumina HISCAN‐SQ platform). We present here the detailed mRNA sequencing workflow (pipeline) customized to provide the full complement of mRNA data (transcriptome) for multiple brain regions over the time course of the SNI protocol. In our initial analysis we focused on the assessment of the transcriptome of the thalamus, which is an important center of pain processing in the brain. We focused in particular on the 21 day time point, which is when the mechanical allodynia reaches its peak and therefore the most severe alterations in transcription are expected. In rats experiencing chronic pain relative to controls, we observed a reduction in the expression of several neurotransmitter receptors, such as CHRM4, CHRNB3, CHRNA5, CHRNB4, CNR1, ADORA2A, GPR85, BDKRB2, MC5R, DRD1, DRD2, and associated G proteins, such as GPR85, GNAT2, GNB3, GNG7, as well as the neurotransmitter TAC1. These findings suggest a global desensitization of neural signaling in the thalamus corresponding to chronic pain. Analysis of all time points and brain regions is underway and will provide a comprehensive picture of gene expression changes during the onset and progression of chronic pain.Research was conducted in compliance with the Animal Welfare Act, and all other Federal requirements. The views expressed are those of the authors and do not constitute endorsement by the U.S. Army.Support or Funding InformationThis work was supported by funding from the United States Army Medical Research and Material Command.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Antimutator Activity of the Nudix Hydrolases from E. coli
Authors: Hynes Thomas William , O'Handley Suzanne Freeman 
Published: June 2021

Enzymes of the Nudix Hydrolase superfamily are characterized by the ability to hydrolyze substrates containing nucleoside diphosphate linked to some moiety x, hence the acronym Nudix. Within the Nudix superfamily, MutT is an established antimutator. This functionality is a point of debate for some of the other members of the superfamily. Experiments to analyze antimutator activity were carried out on the 13 Nudix hydrolase E. coli knockouts. Overnight cultures of each E. coli knockout were grown and plated on LB‐agar containing Streptomycin or Nalidixic Acid, or diluted a million‐fold and plated on LB‐agar or LB‐agar containing Kanamycin (knockouts all contain Kanamycin resistance markers). After ~16 hours incubation at 37 °C, colonies were counted. The MutT knockout demonstrated the ability to gain antibiotic resistance to Streptomycin and Nalidixic Acid as observed by many viable colonies on all plates. The other Nudix hydrolase E. coli knockouts grew on LB‐agar and in the presence of Kanamycin but not in the presence of Streptomycin or Nalidixic Acid. These results indicate that MutT from E. coli is in fact an antimutator, as previously established, but the other Nudix hydrolases from E. coli are not antimutators. Complementation studies with Nudix hydrolases from M. tuberculosis will be carried out to determine which, if any, are antimutators comparable to MutT from E. coli.Support or Funding InformationRIT College of Science FEAD grant (SOH)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Redox Regulation of FGF21 in an Obese “Stress‐less” Mouse Model
Authors: Roberts Aaron , Amos Debbie , Santanam Nalini 
Published: June 2021

BackgroundOxidative stress plays a key role in the metabolic syndrome including Type 2 Diabetes and Obesity. It is implicated that oxygen‐derived free radicals alter the function of metabolic tissues thus altering glucose and lipid homeostasis. Catalase is an antioxidant enzyme that helps to catabolize hydrogen peroxide generated by superoxide dismutation which subsequently reduces oxidative stress. The protein hormone Fibroblast Growth Factor 21 (FGF21) plays regulatory roles in lipid and glucose metabolism, and more recently, has been shown to be a novel regulator of oxidative stress giving it clinical significance in the context of Type 2 Diabetes and Obesity. FGF21 expression has been shown to increase under higher oxidative stress conditions.HypothesisWe hypothesized that excess catalase expression would deter oxidative stress mediated metabolic regulation in adipose tissue as exhibited by changes in FGF21 expression levels and insulin sensitivity.MethodsTo test this hypothesis, male control C57Bl6, Catalase transgenic (Cat‐tg) mice that expressed 3–4 fold excess catalase, as well as the newly engineered Bob‐Cat mice, which is a hybrid of Cat‐tg and the leptin resistant obese Ob‐Ob mice (study approved by MU IACUC), were fed normal chow (NC), 45% fish oil (OM3), or 45% high fat (HFD) diet for 8 weeks. Weekly body weights and food consumption were measured. ECHO‐MRI was used to analyze body fat and lean mass. HOMA‐IR was calculated based on fasting glucose and plasma insulin levels. Changes in mRNA expression of key genes regulating metabolic homeostasis (FGF‐21 and Nrf2) were measured in adipose tissue. ANOVA was used for statistical comparisons using GraphPad Prism.ResultsThe HOMO‐IR data showed that the Cat‐tg mice became insulin resistant while on the HF diet, while the C57 and Bob‐Cat mice maintained their insulin sensitivity although fat mass increased on the high fat diet across all three genotypes. Gene expression data showed a significant increase in FGF21 and Nrf2 in the Bob‐Cat mice that were on the HF and OM3 diets. There was a negative correlation between Nrf2 and FGF21.ConclusionThe results of the gene expression data and insulin homeostasis in the presence of excess antioxidants (catalase) show that proper redox balance is necessary for the signaling and function of FGF21. The correlation in FGF21 expression in the HF and OM3 diets of the Bob‐Cat and the maintenance of insulin sensitivity indicate that FGF21 acts to minimize the effects of poor nutritive conditions.Support or Funding InformationDepartment of Biomedical Sciences, Joan C. Edwards School of Medicine, Marshall University, Huntington, WVThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.