bluuebunny/crossref_metadata_2025_split huggingface dataset:
The role of NFκB in pentagalloyl glucose inhibition of TNF‐α‐induced CXCL1 expression and induction of apoptosis‐related genes in racially different triple negative breast cancer cells.
Authors: Mendonca Patricia , Soliman Karam F.A. 
Published: April 2020

Drug resistance is the leading cause of breast cancer‐related mortality in women, and triple negative breast cancer (TNBC) is the most aggressive subtype. CXCL1/GRO‐α has been identified as a pro‐inflammatory chemokine overexpressed in breast cancer, promoting angiogenesis and having an increased potential to metastasize. The current study investigated the effect of the naturally occurring polyphenolic compound pentagalloyl glucose (PGG) in TNF‐α‐induced CXCL1 expression in MDA‐MB‐231 (Caucasian) and MDA‐MB‐468 (African American) TNBC cells. In addition, PGG apoptotic effect was evaluated through the study of apoptosis‐related genes in both cell lines. Results showed that PGG inhibited TNF‐α‐stimulated CXCL1 expression in the transcription and protein level in Caucasian and African American TNBC cell lines. Through mRNA quantification analysis, data showed that PGG downregulated IKBKE (expressed in 60.4% of the breast cancer tumors) and MAPK1/ERK2 (expressed in tumor tissues with 5 to 10‐fold increase) expression in both cell lines. Moreover, PGG was more effective in inhibiting IKBKE expression in MDA‐MB‐231 compared to MDA‐MB‐468 (9 and 2‐fold inhibition, respectively). Also, PGG reduced MAPK1 gene expression in 3‐fold in Caucasian and 6‐fold in the African American cells. Gene transcription studies showed that PGG induced expression of the pro‐apoptotic genes BNIP3, BNIP3L, GADD45A, TP53BP2, and RIPK2, which are critical initiators of the intrinsic and extrinsic apoptotic pathways. Although PGG increased the expression of apoptosis‐related genes in both cell lines, there was a significantly higher expression in MDA‐MB‐231 cells (5 to 14.7‐fold) compared to MDA‐MB‐468 cells (2 to 5.8‐fold). Furthermore, BAK1 gene expression was only induced in African American cells showing a 35.7‐fold increase after PGG treatment. The obtained data show that PGG inhibited the expression of CXCL1 pro‐inflammatory chemokine and upregulated the expression of specific pro‐apoptotic genes. It was concluded that PGG might have great potential in target therapy for TNBC.Support or Funding InformationThis work was supported by a grant from the NIH/NIMHD grant U54 007582
A Mechanistic Omics‐based Evaluation of Mitochondria‐targeting‐H2S‐donor (AP39) Against Doxorubicin Cardiotoxicity
Authors: Veeranki Sudhakar , Scott David A. 
Published: April 2020

Use of the highly effective broad‐spectrum anti‐cancer chemotherapeutic agent, doxorubicin (Dox), induces Dox‐induced cardiotoxicity (DIC) which leads to mortality and morbidity in pediatric cancer survivors. Dox treatment alters cardiac metabolism and mitochondrial function, however, underlying mechanisms and prevention strategies have not yet been discovered, which limits Dox’s broad clinical utilization. Our data demonstrated that Dox, along with mitochondrial toxicity, depleted both mitochondrial hydrogen sulfide (H2S) and mitochondrial localized H2S producing enzyme, 3‐mercaptopyruvate sulfurtransferase (MPST). As H2S is a cardioprotective signaling molecule, we tested the ability of mitochondria‐targeting‐H2S‐donor, AP39, to protect from the DIC. AP39 treatment was highly effective in mitigating the DIC in mouse models. Further, the combined use of RNAseq, targeted metabolomics and 13C‐labeled‐glucose‐isotope tracing approaches in a cardiomyoblast cell line revealed that AP39 conferred protection against dox‐toxicity by reversing key transcriptional and metabolic features. Moreover, there were improvements in mitochondrial antioxidant status, H2S presence, and quality control. Thus, AP39 is instrumental in protecting mitochondria and promoting stress‐resistant metabolism against DIC. In summary, the integrated omics‐based approach revealed that AP39 confers potent cardioprotection against DIC.Support or Funding InformationThe study was supported by the funds from the American heart association award# 17SDG33670578 to Dr. Veeranki. Dr. Veeranki is also supported by NIH grant HL131782 to Douglas Andres (D.A.) and Jonathan Satin (J.S.) (University of Kentucky). We thank Bryana Levitan (University of Kentucky) for the help with Echocardiography. We thank D.A. and J.S. for sharing some reagents. We thank Markey cancer center RM SRF (supported by NCI Cancer Center Support Grant (P30 CA177558)) for the help with OCR assays. The Vevo 3100 was supported by the Saha Cardiovascular Research Center.
TRPC6 deficiency or feeding a high fat diet impairs ventilatory responses to hypercapnia
Authors: Moak Sydney P. , Wang Zhen , da Silva Alexandre A. , Dai Xuemei , Hall John E. , do Carmo Jussara M. 
Published: April 2020

Transient receptor potential cation channel C (TRPC) is essential for hypoxic pulmonary vasoconstriction response and alveolar gas exchange. We previously showed that whole body TRCP6 deficient (TRPC6 KO) mice are obese and resistant to the anorexic effects of leptin, a hormone that modulates ventilatory control. In the present study we investigated ventilatory responses to hypercapnia (7% CO2) in male and female 22‐week‐old TRPC6 KO and B6/129 control mice fed normal or high fat diet (HFD) since weaning (n=5 per group/sex/diet). Pulmonary ventilation (VE), tidal volume (VT) and respiratory frequency (FR) were measured using the plethysmography method. TRPC6 KO mice were heavier than control mice when fed normal diet (46.2±1.9 vs. 36.6±2.2 g in males and 34.0±1.4 vs. 26.9±1.3 g in females). However, when fed a HFD, male and female TRPC6 KO and B6/129 mice had similar body weight (43.6±0.4 vs. 47.5±1.4 g and 40.1±0.6 vs. 40.7±2.5 g, respectively). Compared to B6/129 controls male and female TRPC6 KO mice fed normal diet had similar VE (1817± 233 vs. 2139±135 and 2146± 404 vs. 2943±421 ml/min/kg) and VT (10.9±0.6 vs. 9.9±0.6 and 17.0±3.4 vs. 14.6±0.8 ml/kg), but lower FR (130±9 vs. 217±14 and 127±5 vs. 201±26 breaths/min) at room air. During hypercapnia, only male TRPC6 KO mice exhibited lower VE (3631± 493 vs. 6003±698 ml/min/kg), which was due to lower FR (235 ±16 vs. 305±19 breaths/min) and VT (15.3±1.0 vs. 19.5±1.4 ml/kg) responses to 7% CO2. Although female TRPC6 KO mice also showed lower FR to hypercapnia compared to B6/129 controls (242±6 vs. 313±22 breaths/min) they exhibited higher VT (28.4±3.0 vs. 25.8±2.0 ml/kg), thus resulting in similarly increased VE (6914±847 vs. 8119±956 ml/min/kg). Male and female B6/129 mice fed a HFD had significantly reduced VE responses to 7% CO2 (4020± 332 vs. 6003±698 and 5140± 194 vs. 8119±956 ml/min/kg, respectively) that were mainly caused by reduced VT (14.4±1.3 vs. 19.5±1.4 and 18.8±0.6 vs. 25.8±2.0 ml/kg). No further reduction in in ventilatory responses to hypercapnia were observed in male or female TPRC6 KO mice fed a HFD. These results suggest that TRPC6 plays an important role in ventilatory responses to hypercapnia, albeit more profound in males than females, and that HFD significantly reduces ventilatory responses to hypercapnia in both male and female control mice, but not in TRPC6 KO mice. The mechanisms involved in the reduced ventilatory responses are still unclear but may involve reduced sensitivity to leptin evoked by HFD or TRPC6 deficiency.Support or Funding InformationNIDDK 1RO1121411, NHLBI‐PO1HL51971, NIGMS P20GM104357 and U54GM115428
Adaptations to cursoriality and digit reduction in the forelimb of the African wild dog (Lycaon pictus)
Authors: Koshy Rahul , Alwiel Ryan , Adrian Brent , Grossman Aryeh , Smith Heather F. 
Published: April 2020

The African wild dog (Lycaon pictus), a rare canid native to southern and eastern Africa, lives and hunts in complex social groups. It is currently classified as Endangered on the IUCN Red List of Threatened Species, with approximately 1400 mature individuals present in the wild. L. pictus is distinct among canids in lacking a digit I or “dew claw” and in its use of exhaustive predation to capture its prey instead of speed, strength, or stealth. These behavioral and morphological traits suggest a potentially unique set of adaptations. This study is the first detailed anatomical description and functional analysis of the forelimbs of L. pictus.The forelimb myology of L. pictus is generally similar to other canids; however, it shows some notable exceptions. While a complete digit I is absent in L. pictus, a vestigial first metacarpal is present, resulting in changes to insertions of mm. extensor digiti I et II, abductor (et opponens) digiti I, and flexor digiti I brevis. M. triceps brachii caput laterale had a larger origin in L. pictus compared to other carnivorans, and m. triceps brachii caput longum had an additional accessory head. Electromyographic (EMG) studies have shown this muscle is active during the stance phase of trotting and galloping and is important for storing elastic energy during locomotion. Mm. anconeus, brachialis, and flexor carpi ulnaris caput ulnare all had more extensive origins in L. pictus than other canids. In domestic dogs, these muscles have a relatively high proportion of Type 1 (slow‐twitch) fibers and are important for posture and elbow stability. Statistical analyses of the quantitative muscle data reveal similar proportions of antebrachial muscle groups to other large‐bodied carnivorans. However, L. pictus has relatively smaller wrist rotators than any other included taxon, suggesting a need for stability over rotatory movement in the carpus of L. pictus.We interpret these differences in size and attachments of muscles in L. pictus compared to other canids as adaptations for long distance running in this highly cursorial species, likely important for exhaustive predation. Absence of a digit I in L pictus, typically used to reduce torque during quick turns and for lightly gripping onto objects, may be related to a reduced need for gripping and quick agile movements in its cursorial lifestyle.Support or Funding InformationThis project was funded by Midwestern University, including a Kenneth A. Suarez Summer Research Fellowship to RK.
Role of TRPV4 mechanosensing in foreign body response and giant cell formation
Authors: Rahaman Shaik O. , Arya Rakesh K. , Goswami Rishov 
Published: April 2020

Implantable biomaterials and medical devices are used in millions of procedures each year worldwide. However, the implantation of these devices leads to the development of a foreign body response (FBR), a chronic inflammatory condition that can ultimately lead to implant failure, which may cause harm to or death of the patient. Despite decades of research, the mechanisms underlying the FBR remain poorly understood, as neither the materials from which these implants are made nor their chemical properties can explain triggering of the FBR. Hallmarks of the FBR include activation of macrophages at the tissue‐implant interface, formation of destructive foreign body giant cells (FBGCs), and development of fibrous tissue that encapsulates the implant. Here, we report that genetic ablation of TRPV4, an ion channel in the transient receptor potential vanilloid family, and which is a mechanosensor, is possibly the mediator of FBR. Specifically, we found that: 1) Trpv4 deletion in mice prevented macrophage accumulation, FBGC formation, and collagen accumulation in a subcutaneous implantation model; 2) the severity of the in vivo macrophage accumulation at the tissue‐implant interface was dependent on the stiffness of the implant; 3) genetic ablation of TRPV4 blocked macrophage adhesion and spreading on stiff matrix in bone marrow derived macrophages; and 4) genetic deficiency of TRPV4 blocked cytokine‐induced FBGC formation, which was restored by lentivirus‐mediated TRPV4 reintroduction. Furthermore, we found that: 5) TRPV4 activity (calcium influx) was augmented in response to a biomechanical stimulus; and 6) loss of TRPV4 activity reduced internal stiffness in macrophages as determined by Atomic Force Microscopy analysis. Taken together, these results suggest an important, previously unknown role for TRPV4 in FBR and giant cell formation.Support or Funding InformationFunding: This work was supported by NIH (1R01EB024556‐01) and NSF (CMMI‐1662776) grants to Shaik O. Rahaman.
A Tool to Predict Affinity and Specificity of GAG Binding to Proteins
Authors: Desai Umesh R. 
Published: April 2020

Glycosaminoglycans (GAGs) are unusual biopolymers that interact with a large number of proteins that reside intracellularly, extracellularly or on cell membranes. These include growth factors, chemokines/cytokines, serine proteases, serpins, metalloproteases, histone acyltransferases, integrins and other cell membrane receptors. Antithrombin is the only serpin believed to bind GAGs with high specificity. However, our computational algorithm is beginning to challenge this prejudicial assumption. In fact, our computational tool shows that GAG display selectivity from both – chemical as well as biological – perspectives. Alternatively, distinct group of GAG sequences selectively recognize a target protein from the horde of sequences (chemical specificity) and distinct binding sites on a protein uniquely recognize a group of GAG sequences (biological specificity). This understanding has come about from the computational tool we have developed to study GAG – protein interactions. The use of this novel combinatorial virtual library screening (CVLS) tool to predict affinity and specificity of GAG sequences for target proteins will be presented. In principle, the tool could be applied to any protein – GAG system of interest to identify the precise GAG sequence that offers high selectivity and high affinity.Support or Funding InformationU01 CA241951; K12 HL141954; R25 HL128639; R01 HL090586
Genomic Profiling and Physiological Approaches to Understand Aquaporins and their Role in ROS Signalling within Skeletal Muscle
Authors: Staunton Caroline Amy , Vasilaki Aphrodite , McArdle Anne , Jackson Malcolm , Barrett-Jolley Richard 
Published: April 2020

Aquaporin (AQP) channels are a family of integral membrane proteins shown to aid the movement of hydrogen peroxide (H202) across membranes
[1]. Their role in skeletal muscle is of key importance since intracellular generation of H202 and other reactive oxygen species (ROS) occurs in response to muscle contractions. These ROS initiate signalling processes and activate of a number of transcription factors
[2,
3]. During ageing, excessive levels of ROS have pathological effects, inducing muscle degradation, loss of neuromuscular junction integrity and loss of neuromuscular transmission.Our studies
[4], as well as those of others, has identified the sources, sinks and functions of H202 in muscle but exactly how H202 is transported from its site of origin within muscle has never been examined. We hypothesize that (AQPs) are the primary means of regulating H202 movement through skeletal muscle membranes. This study has therefore examined expression of AQP isoforms in muscle and how they may change with age.RNA transcripts from Tibialis Anterior (TA) muscle from control adult (6–8 months), and old (26month) mice, and from mice following peroneal nerve crush, were examined by RNAseq (150bp‐read depth >280 M clusters per lane, n=5). Individual muscle myotubes were also treated pharmacologically with aquaporin inhibitors and transfected with AAV6‐HyPer2 to determine the role of different AQP isoforms on intracellular H202 content and H20 permeability was determined using the swell assay
[5,
6].RNAseq data identified 8 AQP mRNA transcripts present in TA muscle. A decrease in AQP4 and AQP1 and an increase in AQP3 contents were found following nerve crush (n=5, P<0.01) and this was confirmed by immunological techniques. The localisation and roles of these aquaporins is currently being investigated to determine whether they have protective functions or contribute to the age‐related decline skeletal muscle.An exponential increase in intracellular H2O2 levels was recorded when AAV6‐Hyper2 transfected muscle fibres were exposed to 10μM H2O2. Pre‐treatment of muscle cells with AQP blockers HgCl2 (30μM), TEA (3mM) or Bumetanide (5μM) substantially decreased the rate of increase in intracellular H202 content. These data demonstrate the ability of aquaporins to regulate skeletal muscle H2O2 permeability. Challenging cells with an 80mOsm solution in conjunction with pharmacological AQP1 inhibitors also demonstrated a significant reduction in the rate of increase in cell volume.In summary, these data demonstrate a potential important role for AQPs in muscle. Further work using AAV‐shRNA to knockdown specific AQPs is being utilised to identify the major AQPs that function in skeletal muscle.Support or Funding InformationThis study was supported by the MRC and NIA (AG051442).
A Novel Role for the ZNF16 Protein in rRNA Transcription
Authors: George Chelsea L. , Diaz-Martinez Laura A. 
Published: April 2020

The zinc finger protein 16 (ZNF16) has been reported to be involved in megakaryocytic and erythroid differentiation of pluripotent K562 leukemia cells. Analysis of the role of ZNF16 in these cells identified functions in cell cycle regulation and c‐KIT transcription. However, ZNF16 has ubiquitous expression in multiple tissues, thus suggesting that it might have additional functions. We explored potential roles for ZNF16 using a battery of assays to test for localization, roles in cell cycle, apoptosis and DNA repair, among others. Endogenous ZNF16 localizes to the nucleus and is enriched in the nucleolus. Interestingly, exogenous ZNF16 showed two patterns of localization depending on the tag used. ZNF16 tagged with a single EGFP (Enhanced Green Fluorescent Protein) showed nuclear localization while ZNF16 tagged with three EGFPs was mainly localized to the nucleoli and other subnuclear structures. Based on its nucleolar localization, we explored potential roles for ZNF16 in nucleolar structure, response to nucleolar stress, rRNA transcription, DNA damage, and DNA repair. Preliminary results hint at a role for ZNF16 in rRNA transcription. These results reveal a novel role for ZNF16 in basic cellular processes, consistent with its ubiquitous expression in human tissues. Further characterization at the cellular and molecular level will offer insight on the function of this poorly characterized gene.Support or Funding InformationUTEP‐URI program; BD Biosciences Cancer Research Grant; Murdock Foundation and Gonzaga Start‐up funds.
TNFα Exposure Decreases Mitochondrial O2 Consumption in Motor Neurons
Authors: Heino Amanda Denise , Yap Jane Denise , Brown Alyssa Denise , Rana Sabhya Denise , Fogarty Matthew J. , Mantilla Carlos B. , Sieck Gary C. 
Published: April 2020

Neuroinflammation triggers a homeostatic response that is mediated by pro‐inflammatory cytokines such as TNFα. We hypothesize that in a motor neuron‐like cell line, NSC‐34, TNFα decreases mitochondrial O2 consumption. NSC‐34 cells were differentiated by serum depletion (~24 h) until neurite structures (>50 μm) were observed. The cells were then exposed to TNFα (20 ng/ml) for 24 h. To determine mitochondrial volume density, the cells were loaded with MitoTracker Green and imaged in 3D using a Nikon A1R confocal microscope (60x oil immersion 1.4 NA). After treatment with TNFα, mitochondria appeared fragmented but mitochondrial volume density increased by ~50%, as compared to control untreated cells. A Agilent Seahorse XF Analyzer was used to determine the O2 consumption of treated and untreated cells during a stress test that determines basal respiration, ATP production and maximum respiration. O2 consumption was normalized to mitochondrial volume density in both treated and untreated groups. After TNFα exposure, basal respiration, ATP production and maximum respiration decreased per mitochondrion. However, overall O2 consumption of the cells was unchanged after TNFα exposure due to an offsetting increase in mitochondrial volume density. Using MitoSox, we also measured reactive oxygen species (ROS) formation in treated and untreated NSC‐34 cells. ROS is a byproduct of mitochondrial O2 consumption, and can cause cellular damage. After TNFα exposure, ROS formation increased compared to untreated cells. We conclude that mitochondrial biogenesis and an increase in mitochondrial volume density represent a homeostatic mechanism to reduce O2 consumption the per mitochondrion, thereby damping ROS formation while maintaining overall cellular O2 consumption and ATP production.Support or Funding InformationSupported by NIH grant AG44615 and HL105355.
Defining how viruses manipulate lipid phosphoinositides through activation of PI4P kinases to mediate viral replication
Authors: Burke John E. , McPhail Jacob A. , Rathinaswamy Manoj , Jenkins Meredith L. 
Published: April 2020

Phosphoinositide kinases are central regulators of myriad cellular processes, and their misregulation is causative of multiple human diseases. We are currently focusing on the structural basis for the regulation of phosphoinositide kinases, specifically the Golgi localised phosphatidylinositol 4‐kinase beta (PI4KB). We have identified a previously uncharacterised protein binding partner of PI4KB, the GTPase regulatory protein c10orf76
1, and revealed novel insight into an unexpected PI4K positive feedback loop. Using a combined hydrogen deuterium exchange mass spectrometry (HDX‐MS) and structural biology approach we have defined the molecular basis of regulation by the protein c10orf76, revealing novel insight into the role of these PI4KB in viral replication. This has revealed key insight into unexpected feedback loops with Ras superfamily GTPases. Overall, this approach has provided a novel toolbox of engineered mutations to tease out the molecular basis of regulation of phosphoinositide kinases in both homeostasis and disease, and has revealed novel opportunities for the development of small molecule therapeutics for multiple pathogenic viral infections.Support or Funding InformationResearch was supported by the Canadian Institute of Health Research (CRN‐142393), and salary award for the CIHR new investigator and MSFHR scholar programs.