bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of Primary Cilia in Cell Migration
Authors: Ganesh Jyothsna , Miskevich Frank 
Published: November 2023

Cellular migration is a common activity for many different types of cells. If cellular migration is disrupted, cells may be unable to repair damaged tissue (wound repair), migrate appropriately as in cancer, or correctly signal their cellular state. In several situations, cilia have been implicated in helping to determine the direction of cellular migration. As primary cilia express a number of distinct receptors for particular hormones, these organelles have the required machinery to influence cellular behavior. We are using ARPE‐19 cells, a human retinal pigment epithelial cell line capable of forming cilia in culture, as an assay for cellular motility. Using a 3D model of cellular migration (matrigel invasion chambers) and a 2D scratch assay for cellular motility, we are testing various reagents for their ability to alter RPE cell motility. Preliminary results suggest that a number of agents which alter ciliary function, particularly protein kinase A modulators, are able to inhibit the migratory ability of ARPE cells. These results suggest that protein kinase A may be a key link between ciliary signaling and the direction of cellular migration.Support or Funding InformationUniversity of Michigan Graduate Student Research Assistantship; University Research and Creative Activities grant
Prolyl isomerase Pin1 regulates the stability, transcriptional activity and oncogenic potential of Brd4
Authors: Hu Xiangming 
Published: November 2023

The multi‐functional Brd4 belongs to the two tandem bromodomains and an extra terminal (ET) domain‐containing family proteins (BETs), which have emerged as important epigenetic modulators for gene transcription and cancer development. Brd4 has been implicated in the development of hematological malignancies, including mixed‐lineage leukemia (MLL), multiple myeloma, and adult T cell leukemia. While accumulating evidence indicates that Brd4 has a pivotal role in the tumorigenesis by activating the genes responsible for cell proliferation and survival, how Brd4 itself is regulated in cancer remains elusive.In this study, we report that the stability and functions of Brd4 are positively regulated by prolyl‐isomerase Pin1. Pin1 directly binds to phosphorylated Brd4 and enhances Brd4's stability by inhibiting its ubiquitination. Pin1 also catalyzes the isomerization of Brd4 and induces its conformational change, which promotes its interaction with CDK9 and increases Brd4's transcriptional activity. Substitution of Brd4 with Pin1 binding‐defective mutant reduces Brd4's stability, attenuates Brd4‐mediated gene expression by impairing its interaction with CDK9, and suppresses cancer cell proliferation, migration and invasion, and tumor formation.Our results identify Brd4 as a new target of Pin1 and suggest that interfering with their interaction could be a potential therapeutic approach for cancer treatment.Support or Funding InformationUIUC (to L.F.C.) and NIH grants DK085158 and CA179511 (to L.F.C.)
Characterization of human SH2 domain‐containing protein 4A (SH2D4A) in HeLa cells
Authors: Tamborlin Leticia , Pereira Karina Danielle , Meneguello Leticia , de Proença Andre Ricardo Gomes , Luchessi Augusto Ducati 
Published: November 2023

The SH2 domain‐containing protein 4A (SH2D4A) has a single Src homology region 2 domain (SH2 domain) and a proline‐rich stretch on its C‐terminal. In humans, SH2D4A gene is located on chromosome 8p close to other genes associated with cancer. This region is deleted in patients with hepatocellular carcinoma (HCC) with poor outcomes, suggesting that depletion of SH2D4A could be involved with tumor progression. Besides, SH2D4A inhibits the STAT3 transcription factor activity mediated by interleukin 6 (IL‐6) signaling pathway, which interacts physically with STAT3, inhibiting the transcription of genes involved with cell proliferation, survival, and tumor development. Five transcripts corresponding to SH2D4A gene are described (variants 1, 2, 3, X1 and X2). The transcript variants 1 and 2 are predicted to encode the same isoform (isoform A), which has 454 amino acids and is considered canonical. The transcript variant 1 presents an upstream Open Reading Frame (uORF). Thus, this study aimed to characterize SH2D4A gene and protein in a model of tumor cells (HeLa cells), looking for new evidences to evaluate the involvement of this protein in development and tumor progression. First of all, we performed amplification of the cDNA by RT‐PCR and subsequent cloning. Thereby, it was possible to identify the transcript variants 1, 2 and X1 of SH2D4A, which was confirmed by sequencing of the samples. Analyzes of the sequenced samples revealed some mutations in SH2D4A gene, among them, the most significant was observed in the start codon, which results in the use of an alternative start codon to production of the protein. In addition, the presence of the uORF in SH2D4A variant 1 downregulates the protein production from transcript variant 1 in transfected cells. In fact, a depletion of the uORF increased the protein content compared to original variant 1. Our results suggest these mutations in SH2D4A gene and the presence of the uORF in transcript variant 1 may contribute to the abnormal characteristics observed in this type of cells.Support or Funding InformationSão Paulo Research Foundation (FAPESP)
LPIN1 promotes epithelial cell transformation and mammary tumourigenesis via enhancing insulin receptor substrate 1 stability
Authors: Choi Hong Seok 
Published: November 2023

LPIN1, a phosphatidic acid phosphatase enzyme, dramatically influence tumor progression, however, their mechanism of action is still poorly understood. Here, we demonstrate that LPIN1 enhances the tumour‐promoting function of IRS1 by controlling IRS1 stability. LPIN1 interacts with IRS1 in an IGF‐1‐dependent signalling pathway and inhibits its serine phosphorylation, thereby eliminating ubiquitin‐dependent degradation of IRS1 via proteasomal and lysosomal pathways. Consequently, LPIN1 overexpression increases IRS1 abundance and enhances IRS1's ability to induce epithelial cell proliferation and mammary tumourigenesis. By contrast, depletion or inhibition of LPIN1 in breast cancer cells leads to a decreased IRS1 level, which subsequently inhibits the RAF1‐mediated signalling pathway as well as AP‐1 transcriptional activity. In the syngeneic 4T1 breast cancer model, LPIN1 overexpression increased tumour development. Consistent with these observations, LPIN1 levels were positively correlated with IRS1 expression in human breast cancer. Thus, our results indicate that LPIN1 may be a promising drug target for anticancer therapy.Support or Funding InformationThis research was supported by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute, funded by the Ministry of Health & Welfare, Republic of Korea (grant number: HI14C2004).
Withaferin A suppresses and inhibits hepatocellular carcinoma‐cell proliferation and tumor burden via activation of ERK1/2‐ELK1‐RSK‐DR5 axis
Authors: Kuppusamy Panjamurthy , Nagalingam Arumugam , Drachenberg Cinthia , Sharma Dipali , Saxena Neeraj K 
Published: November 2023

Hepatocellular carcinoma (HCC) is the fifth most common highly lethal cancer and the third most common cause of cancer‐related mortality in the world. Available therapies are often limited by high toxicity, lower efficacy, therapeutic resistance and therapy‐related morbidity. Liver resection, ethanol injection, radiofrequency ablation, and chemoembolization have been used successfully for the local management of HCC; however, no single cytotoxic‐chemotherapeutic‐agent has been proven effective for systemic treatment. Therefore, more effective therapeutic strategies are needed to combat liver cancer related mortality. Withaferin A (WA) is a bioactive compound derived from Withania somnifera has been safely used for centuries in Indian‐Ayurvedic‐medicine against various ailments. For the first time, we demonstrated that WA displays substantial inhibitory activity against liver cancer cells in vitro and tumor burden in vivo in HCC‐cell‐xenograft‐nude mice and DEN induced HCC in C57BL/6.MethodsTherapeutic efficacy of WA was assessed by determining cell viability of HCC cell lines, HepG2 and Huh7, using XTT reduction while apoptosis was quantified by TUNEL assay. Effect of WA on cell migration (spheroid assay), invasion (matrigel invasion), clonogenicity, anchorage independent growth (colony formation on soft agar) were determined. Additionally, in a quantitative real‐time, ECIS (Electric cell‐substrate impedance sensing)‐based assay was performed to evaluate efficacy of WA on migration and invasion of HCC cell lines. The mechanism for the inhibitory role of WA on HCC was determined by immunoblot assay for cleaved caspase 3, Bax, cleaved PARP, phosphorylated ERK, Elk1, RSK1 and DR5 in HCC cells, tumors derived from xenografts in nude mice and tumorous‐livers from DEN treated mice.ResultsWA treatment significantly inhibited HCC cell‐proliferation, migration, invasion, clonogenicity and anchorage‐independent growth. WA treatment significantly induced apoptosis to suppress cell and tumor growth as indicated by increased cleaved caspase 3 activity, cleaved PARP and Bax. Mechanistically, WA induced expression of death receptor 5 (DR5) that triggers apoptosis when overexpressed. Our study revealed that WA inhibits phosphorylation of Stat3 which in turn up‐regulates phosphorylation of Elk1. Inhibition of ERK or Elk1 phosphorylation inhibits WA‐induced DR5 induction, indicating that ERK and Elk1 cooperatively regulate DR5 expression.ConclusionsCollectively, our studies show the efficacy of WA as a novel agent for HCC inhibition and provide mechanistic insight into the important role of ERK‐Elk1 axis in WA‐mediated HCC inhibition.
The cytoprotective effect of chlorogenic acid against oxidative stress‐induced cell damage via the ERK and PI3K/Akt‐mediated Nrf2/HO‐1 signaling pathways
Authors: Jang Miran , Kim Gun‐Hee 
Published: November 2023

This study investigated the effect of chlorogenic acid of Cirsium japonicum on the expression and activity of heme oxygenase‐1 (HO‐1), an enzyme with potent antioxidant properties, as well as the molecular mechanisms involved. Chlorogenic acid markedly upregulated HO‐1 protein expression in PC12 cells. Chlorogenic acid also increased the protein level of transcription factor nuclear factor erythroid 2‐related factor 2 (Nrf2), which regulates HO‐1 expression by binding to the antioxidant response element (ARE) within the HO‐1 gene promoter, in a dose‐and time‐dependent manner. Exposure to H2O2 reduced the expression of HO‐1 in PC12 cells, the addition of chlorogenic acid restored HO‐1 expression protein levels. Moreover, chlorogenic acid decreased the expression of Kelch‐like ECH‐associated protein 1(Keap 1). Chlorogenic acid activated extracellular‐regulated kinase (ERK) and protein kinase B (PKB, Akt) in keratinocytes, while the ERK and Akt inhibitors attenuated chlorogenic acid‐enhanced Nrf2 and HO‐1 expression. The results suggested that chlorogenic acidactivates ERK‐and Akt‐Nrf2 signaling cascades in PC12cells, leading to the upregulation of HO‐1 and cytoprotection against oxidative stress. The phenolics in Cirsium japonicum leaf, particularly, chlorogenic acid can be utilized as effective and safe functional food substances, i.e., natural antioxidants, and may reduce the risk of neurodegenerative disorders.Support or Funding InformationThis work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIP) (No.2016R1A2B4014740)
Undergraduate Research Projects at the Community Colleges
Authors: Hess Brittany A , Fortner Alexia A , Chowdhury Syed A 
Published: November 2023

About 45% of the nation's undergraduates experience their first post high school science education at a 2 year technical college. However, unfortunately almost none of these colleges have adequate undergraduate research opportunities for these students. Science education, by definition is, “the learning of scientific phenomena through investigation or research”. Research enhances our mental capacities and engages students in critical thinking. In a community college or a technical college setting, it is extremely difficult and also unlikely for students to spend a significant amount of time conducting research. Along with this, most instructors do not get enough time to write grants or participate in research when they have a heavy teaching load and often times serve on various committees. However, we have now created new opportunities for our undergraduate science students who show their interst in research. These newly added projects are concise, need little mentoring, and require minimal or no costs. With the help of our instructors and using current lab facilities, we have made dozens of projects readily available for our students. Any student can pick a project from those offered and get involved in simple yet extremely relevant research on campus. For instance, we have ‘testing attention and memory related brain functions’ which uses less expensive and much more portable equipment known as ‘PsychoToolKits’. We have also included, ‘Biopac’ to perform challenging experiments related to heart function, brain waves and muscle activities, or in order to examine the ‘behavior of microbes’ in multiple altered environments. Students have shown great interest in these projects and seem to enjoy investing time to understand and work through these labs thoroughly. We recommend that community as well as many other 2‐year colleges get involved in creating undergraduate research projects and we would love to see them offer these opportunities to students who are eager to study how the methods of planning, conducting, and gathering relevant information to research from numerous sources and then being able to analyze and present the data to their peers. Students will greatly benefit from being involved in these newly incorporated projects.Support or Funding InformationDepartment of Science, Saint Louis Community College‐Wildwood
NLRP3 Inflammasome Is Critical for Lipopolysaccharide‐Induced Depressive‐Like Behaviors
Authors: Lee Eunju , Jeon Seon‐A , Hwang Inhwa , Yu Je‐Wook 
Published: November 2023

Cerebral inflammation is considered to contribute to the pathogenesis of depressive disorders, but the molecular target to intervene inflammation‐mediated depressive symptoms is still poorly elucidated. Recent studies intriguingly proposed the implication of NLRP3 inflammasome signaling in the depressive‐like disorders, although the molecular basis by which NLRP3 inflammasome activation contributes to depression is unclear. In this study, we examined lipopolysaccharide (LPS)‐induced depressive‐like behaviors in wild‐type and Nlrp3‐deficient mice. The levels of proinflammatory cytokines in the brain homogenate were not significantly different between wild‐type and Nlrp3‐deficient mice after LPS administration. However, significant reduction of active caspase‐1 was observed in the hippocampus of Nlrp3‐deficient mice brain compared to wild‐type mice. Of interest, LPS administration to mice caused a cerebral induction of Ido mRNA in a NLRP3‐dependent manner. Supporting this data, NLRP3 inflammasome‐activating stimulations, but not interleukin‐1β treatment, triggered a significant induction of IDO in mixed glial cells. Collectively, our findings suggest that IDO is a potential downstream mediator of the NLRP3 inflammasome in the progression of depressive‐like behaviors.Support or Funding InformationThis work was supported by the Bio & Medical Technology Development Program of the NRF funded by the Korean government, (2015M3A9B6073856)
Pseudomonas aeruginosa Attenuates Inflammasome Activation via Quorum Sensing‐Dependent Mechanism
Authors: Yang Jungmin , Lee Kang‐Mu , Park Sangjun , Yoon Sang‐Sun , Yu Je‐Wook 
Published: November 2023

Quorum sensing (QS) is a bacterial communication mechanism that promotes coordinated adaptation by triggering expression of a wide range of genes. QS is thought to strongly contribute to the virulence of Pseudomonas aeruginosa, an opportunistic bacterial pathogen, but the molecular effect of QS on host inflammasome defense is completely unknown. Here, we present evidence that P. aeruginosa dampens host inflammasome signaling by QS‐related products in mouse bone marrow‐derived macrophages. We demonstrate that QS‐regulated virulence factor pyocyanin and QS autoinducer 3‐oxo‐C12‐homoserine lactone directly suppress NLRC4‐ and NLRP3‐mediated inflammasome assembly and activation. In addition, bacteria‐free secretant from QS‐defective ΔlasR/rhlR mutant, but not from wild type bacteria, induces robust activation of the NLRC4 inflammasome. Extracellular P. aeruginosa‐released flagellin might be responsible for this inflammasome activation by ΔlasR/rhlR secretant, but QS‐dependent secretion of bacterial proteases degrade extracellular flagellin to evade NLRC4 inflammasome activation. P. aeruginosa–secreted proteases also impair inflammasome components and pro‐inflammatory cytokines in the extracellular space. Taken together, our data indicate that QS system of P. aeruginosa facilitates bacteria to evade host inflammasome‐dependent sensing machinery.Support or Funding InformationThis work was supported by the Bio & Medical Technology Development Program of the NRF funded by the Korean government, (2015M3A9B6073856)
Generating a novel cell line with a codon optimized LINE1 element at a single locus
Authors: Russell Brittany , Morales Maria , DeFreece Cecily 
Published: November 2023

Long interspersed element‐1 (LINE1) is the only currently active autonomous retroelement within the human genome. LINE1 replicates itself through a process known as retrotransposition that requires the endonuclease and reverse transcriptase activities of the LINE1 protein ORF2. Further studies characterizing LINE1 endonuclease function demonstrated that LINE1 endonuclease induces formation of double‐stranded breaks (DSBs). Only a small fraction of LINE1‐induced DSBs results in a retrotransposition event, suggesting that the damage from the LINE1 endonuclease activity may be greater than previously considered. While LINE1 mutagenic insertion events have been associated with diseases such as breast and colon cancer as well as muscular dystrophy, the full effect of LINE1 endonuclease upon stability of the genome is undetermined. By inserting a codon‐optimized LINE1 element into a cell line at a unique genomic locus using the FRT system, we will investigate and track the effect of the endonuclease on the genome. Codon optimization of LINE1 will increase expression of the element and will aid in detection of novel retrotransposition events in the presences and absence of drugs. Here we are presenting the cloning of the FRT vector containing the codon‐optimized LINE1 element, the first step of project.Support or Funding InformationThe work presented is supported in part by monies from P20GM103424, TL4GM118968 and 5RL5GM118966‐03 as well as the Louisiana Cancer Research Consortium.
Direct and Specific Interaction of MIF and Bacterial Endotoxin May Play an Important Role in the Pathogenesis of Sepsis
Authors: Cho Thomas 
Published: November 2023

Inflammatory response is triggered in the body as a result of infections by bacteria and other pathogens. Especially, a bacterial outer membrane component, lipopolysaccharide (LPS), is a major virulence factor. LPS activates the secretion of various inflammatory cytokines. Among the secreted cytokines, macrophage migration inhibitory factor (MIF) is a cytokine‐like protein that amplifies inflammation by activating the secretion of other inflammatory cytokines. MIF levels are elevated in severe sepsis and septic shock. MIF−/− mice demonstrated improved survival from sepsis. This implies that MIF plays an important role in sepsis, but the underlying molecular mechanism is unknown.Our recent data demonstrated evidence that MIF dramatically improved CXCR4‐mediated cell responses only when it makes a complex with LPS. Based on this observation, we hypothesized that LPS released during bacterial infection forms a complex with MIF and the complex plays a deteriorating role in the pathogenesis of sepsis. Removal of bacterial endotoxin from recombinant MIF is challenging and it is controversial whether MIF alone or MIF complexed with a binding partner mediates cell responses leading to various inflammation diseases. We noticed that a conventional endotoxin‐removal column failed to separate endotoxin from MIF. This led us to investigate whether MIF and LPS directly bind each other. MIF has one tryptophan (W109) exposed to the solvent. We demonstrated that the intrinsic fluorescence of W109 was quenched when titrated with increasing concentrations of LPS. We confirmed the MIF‐LPS interaction by using isothermal titration calorimetry and determined the affinity (KD) of 2.9 micromolar. We also proved that MIF and LPS formed a complex during protein purification and in an animal model of sepsis. Finally, we identified a LPS binding site located at the trimeric interface of MIF by using NMR.To elucidate potential biological consequences of the MIF‐LPS interaction, we tested whether this interaction alters the effect of each molecule on endothelial barrier function, which is damaged during conditions such as sepsis. Intriguingly, LPS together with MIF revealed less endothelial permeability than LPS alone. MIF alone did not significantly change endothelial barrier permeability. Our findings on the MIF‐LPS interaction could serve as a novel paradigm to better understand physiological functions of this important and enigmatic immune regulator.Support or Funding InformationNIH NIAMS