bluuebunny/crossref_metadata_2025_split huggingface dataset:
Loss of Dsg2 Promotes Tumorigenesis in Pancreatic Cancer Cells through ERK Activation
Authors: Huetz Katharina , Waschke Jens , Spindler Volker 
Published: November 2023

The ability to maintain cell‐cell adhesion is crucial for tissue integrity and organization. Accordingly, loss of cohesiveness plays a critical role in cancer invasion and metastasis. Desmosomes are cell junctions providing strong intercellular adhesive strength and dysregulation of desmosomal constituents was shown to contribute to cancer progression through altered cell signaling pathways.Here, we focused on the cadherin‐type adhesion molecules desmoglein 2 (Dsg2) and desmocollin 2 (Dsc2), building up the desmosomal core. siRNA‐mediated knock down of Dsg2 but not Dsc2 resulted in loss of cell cohesion and enhanced migration of pancreatic adenocarcinoma cells (AsPC‐1). We established stable Dsg2 knockdown cell lines in which loss of Dsg2 caused an increase in cell migration and enhanced invasion through extracellular matrix without influencing cell proliferation.To identify potential pathways regulated by Dsg2, we screened major kinases and detected the activity of ERK to be significantly enhanced in Dsg2‐deficient cells. Consequently, inhibition of ERK phosphorylation in Dsg2 knockdown cells led to normalized migration. Interestingly, loss of Dsg2 was accompanied with reduced levels of the desmosomal adapter protein and transcriptional regulator plakoglobin, whereas other desmosomal molecules were not altered. Consistent with this observation, in initial experiments overexpression of plakoglobin rescued enhanced migration induced by silencing of Dsg2. These results indicate that loss of Dsg2 promotes tumorigenic behavior of pancreatic cancer cells by regulation of ERK activity and the levels of plakoglobin.
Keratin Filaments Regulate Binding Properties of the Desmosomal Adhesion Molecule Desmoglein 3
Authors: Vielmuth Franziska , Loschke Fanny , Magin Thomas M. , Waschke Jens , Spindler Volker 
Published: November 2023

Desmosomes are supramolecular protein complexes required for strong intercellular adhesion. Accordingly, they are most abundant in tissues exposed to pronounced mechanical stress, such as the epidermis. In desmosomes, the interaction with adjacent cells is provided by desmosomal cadherins which are intracellularly linked with the intermediate filament cytoskeleton by several adapter proteins. Using atomic force microscopy (AFM) on living keratinocytes, we recently showed that binding forces of the desmosomal cadherin desmoglein 3 (Dsg3) are not uniformly distributed but are higher at cell borders compared to cell surface areas. In view of the compromised adhesion resulting from intermediate filament loss, we here tested the relevance of keratins, the intermediate filament proteins of epithelial cells, for the binding properties of Dsg3.In cultured human keratinocytes, Dsg3, the adapter molecule desmoplakin and keratin filaments, assembles on the cell surface away from junctions. Murine keratinocytes lacking all keratin filaments (KtyII ko) displayed reduced intercellular adhesion compared to controls (KtyII wt). Surprisingly, the protein levels and membrane localization of Dsg3 was increased in KtyII cells. In line with this, by using AFM, a higher frequency of Dsg3‐mediated binding events was observed in Ktyll ko cells. However, the binding forces of individual Dsg3‐mediated binding events were significantly reduced compared to KtyII wt cells.Our data for the first time show that keratins, potentially by anchorage to the desmosomal complex, regulate binding properties of desmosomal cadherins and therefore provide new insights into the mechanisms of desmosomal adhesion.
Resolution Agonist 15‐epi‐Lipoxin A4 Directs FPR2 to Expedite Healing Phase Post‐Myocardial Infarction
Authors: Kain Vasundhara , Liu Fei , Kozlovskaya Veronika , Ingle Kevin A , Khedkar Santosh , Prabhu Sumanth D , Kharlampieva Eugenia , Halade Ganesh V 
Published: November 2023

Post‐myocardial infarction (MI), overactive inflammation remodels the left ventricle (LV) to cause heart failure, marked with reduced levels of resolution markers such as 15‐epi‐Lipoxin A4 (15‐epi LXA4). However, the role of 15‐epi LXA4 in acute inflammatory response and resolving phase post‐MI is unclear. We hypothesize that 15‐epi‐LXA4 with liposomal fusion (Lipo‐15‐epi‐LXA4) or free 15‐epi‐LXA4 will expedite the resolving phase in post‐MI inflammation. 8 to 12 week old male C57BL/6 mice were subjected to permanent coronary artery ligation. Lipo‐15‐epi‐LXA4 or 15‐epi‐LXA4 (1ug/kg/day) was injected 3 hours post‐MI for day (d)1 or until d5. Mice with no treatment served as MI‐control and mice with no‐MI were used as d0 naïve controls. In‐vitro experiment using peritoneal macrophages showed 15‐epi‐LXA4 activated formyl peptide receptor (FPR2/ALX) and GPR120 on alternative macrophages, but inhibited GPR40 on classical macrophages. The 15‐epi‐LXA4 injected mice displayed reduced LV‐ and lung mass‐to‐body weight ratios and increased fractional shortening at d5 post‐MI. In acute phase (d1), 15‐epi‐LXA4 primes neutrophil infiltration with a robust increase of Ccl2 (FPR2/ALX) (all p<0.05) expression compared to MI‐control group. During resolving phase (d5), 15‐epi‐LXA4 initiated the rapid clearance of neutrophils with persistent activation of FPR2/ALX in both LV and spleen with inhibition of GPR40. 15‐epi‐LXA4 increased transcripts of proresolving markers Arg‐1 and Ym‐1 at d5 post‐MI (all p<0.05) in the infarcted LV. Post‐MI splenic activation and reduced kidney inflammation was marked by an increase in expression of Mrc‐1/CD206 and Ym‐1 (both p<0.05) as well as a decreased level of NGAL and plasma creatinine, respectively in 15‐epi‐LXA4 group. In summary, 15‐epi‐LXA4 initiates the resolving phase early to remit uncontrolled inflammation post‐MI, thereby reduced LV dysfunction.Support or Funding InformationThis work was supported by grants from National Institutes of Health (NIH)‐NCCAM R00AT006704, Michigan Regional Comprehensive Metabolomics Resource Core and UAB start‐up fund to G.V.H
Serum miRNAs as potential biomarkers for early prediction of type 1 diabetes
Authors: Yin Congcong , Weiland Matthew , Li Jia , She Ruicong , Zhou Li , Mi Qingsheng 
Published: November 2023

BackgroundThe incidence of type 1 diabetes (T1D), a T cell‐mediated beta cell destructive autoimmune disease, has been increasing about 3–4% annually for several decades. Individuals at risk for T1D are diagnosed at a late stage when the possibility for disease prevention is absent. Autoantibodies (AA) to islet antigens such as islet antigen (IA)‐2, IA‐2b, and glutamate decarboxylase (GAD65) appear earlier before T1D onset and are used for early T1D prediction. However, the appearance of islet AA marks a relatively late stage of the autoimmune process and therefore is not suitable for early disease intervention. More importantly AA lack causal relationship with the pathogenesis of T1D. Therefore, there is an urgent need for better (increased specificity/sensitivity) and earlier (predating autoantibodies) markers for the prediction of AA development. MicroRNAs (miRNAs) have emerged as an important regulatory factors in pancreatic β‐cell development, homeostasis, function, and in a variety of immune cell development, differentiation and function. Our recent study showed that miRNAs regulate T1D development and serum miRNAs are potential biomarkers for T1D progression in mouse models. Our objective here is to identify specific serum miRNA biomarkers for earlier and better prediction of individuals at risk for T1D in human.MethodWe performed serum miRNA expressions profiles in 35 AA positive (IAA and ICA) non‐T1D subjects and 40 AA negative relative subjects from the Diabetes Prevention Trial‐Type 1 (DPT‐1) cohort, using the TaqMan low‐density arrays. miRNAs with changed expression level were further confirmed by a single TaqMan RT‐PCR.Result9 miRNAs (miR‐146a, miR‐561 and miR‐548a‐3p, miR‐184 and miR‐200a) were down‐regulated and 2 miRNAs (miR‐30c and miR‐487a) are up‐regulated in the serum of AA+ non‐T1D subjects compared to that from AA‐ subjects (two‐sample t‐test P<0.05). In addition, a cluster of five miRNAs (miR‐146a, miR‐197, miR‐193b, miR‐574‐3p, and miR‐561) was identified to clearly separate AA+ subjects from AA‐ subjects with higher sensitivity and specificity ( LASSO logistic regression). miRNA target and pathway prediction analyses revealed that some of these miRNAs are related to immune function and beta‐cell homeostasis and potentially involved in autoimmune processes.ConclusionWe have identified distinct serum miRNA expressions profiles in AA+ subjects compared to AA‐ subjects. These serum miRNAs could serve as potential biomarkers for early prediction of autoimmune processes in individuals at risk for T1D, which need to be further confirmed in the future studies.Support or Funding InformationNIH/NIDDK, XO1DK097691‐01A1 JDRF grants, 17‐2012‐678 and 2‐SRA‐2015‐122‐Q‐R
Lutein, But Not Other Carotenoids, Selectively Inhibits Breast Cancer Cell Growth Through Several Molecular Mechanisms
Authors: Swanson Haley M , Smith Joshua R , Gong Xiaoming , Rubin Lewis P 
Published: November 2023

IntroductionThe dietary carotenoid lutein is critical in eye and brain development and function. Its anticancer effects have been controversial, albeit understudied. We investigated effects of lutein in a wide range of receptor (+) and (−) breast cancer cell lines including difficult‐to‐treat ‘triple‐negative’ breast cancer.Materials/MethodsEight established breast cancer cell lines and primary human mammary epithelial cells (PmEC) were exposed to a concentration range of (1) carotenoids (β‐carotene, lutein, zeaxanthin, lycopene, astaxanthin); and (2) lutein + chemotherapy agents (paclitaxel, docetaxel). Cell proliferation was analyzed by MTT and EdU assays, and soft agar colony formation was assessed. Intracellular reactive oxygen species (ROS), cell cycle, and apoptosis were assayed by flow cytometry. Proteomic arrays and RT‐PCR profiling were used to evaluate changes in cellular protein phosphorylation and targeted gene expression, respectively.ResultsLutein and lutein epoxide, and to a lesser extent lycopene, but not other carotenoids, significantly inhibited breast cancer cell growth and induced cancer cell apoptosis. Lutein had little apparent effect on PmEC cell proliferation or viability. Lutein‐mediated growth inhibition is quantitatively similar to that induced by chemotherapeutic agents tested. Moreover, lutein and chemotherapy act additively‐to‐synergistically to inhibit cell growth. We determined mechanisms for growth inhibition by lutein in triple‐negative breast cancer cells include: induction of G1 cell cycle arrest; increased intracellular ROS production; mutant and wild‐type p53 phosphorylation; and down‐regulation of several anti‐apoptotic proteins, including Bcl2.ConclusionsOur findings demonstrate lutein (in concentrations attained in healthy diets) has anti‐tumor activities in human breast cancer acting via several molecular mechanisms. Lutein supplementation/fortification may be a promising nutriceutical preventive and/or adjunct therapeutic approach in a wide range of human breast cancer subtypes.Support or Funding InformationPartially funded by TTUHSC El Paso Seed Grants (LPR, XG), a Scholarly Activity & Research Program student grant (HMS), and a Summer Accelerated Biomedical Research Internship (JRS).
Functional characterization of Isl2/coupTFIb activation targets in vascular development
Authors: Wang Yi‐Shan 
Published: November 2023

Genetic and signal interaction that regulating the vein identity, intersegmental vessels (ISV) and cardinal vein pluses (CVP) patterning is still not fully understood. We previously identified the transcription factors Islet2 (Isl2) and CoupTFIb required for specification of the vein and ISV growth mediated by Notch pathway in zebrafish. To understand the molecular mechanisms behind the isl2/coupTFIb regulation, we performed the microarray experiments to identify the potential downstream targets. In this study, we focus on the potential activation targets and we confirmed several genes expressed in vessels during early development indicating those potential genes function in vascular development likely downstream of isl2/coupTFIb.We selectively assay the functional relevance of one potential target, signal‐transducing adaptor protein 2 (stap2). Stap2 is identified as an adaptor protein that has been showed functions in a variety of cellular signal pathways in immune system, however, there is no description of stap2 functioning in vascular development so far. Our in‐situ hybridization results showed stap2 mRNA is expressed is expressed in developing vessels from 18S to 48 hpf stages, suggesting its roles in vasculization. Knockdown of stap2 by morpholino injection causes vascular defects, suggesting the role of stap2 in controlling ISV and CVP growth. We confirmed the specificity of stap2 MO knockdown by using 2nd MO interfere block splicing site, examining knockdown efficiency, and expressing stap2 mRNA to rescue stap2 morphants. AO staining and TUNEL assay showed that vascular defects do not caused by cell death, but due to the impairment of migration and/or proliferation by examining ISV integrity and ISV cell numbers in Tg(fli:neGFP; kdrl:mCherry) fish. To test molecular mechanisms of vascular defects in stap2 morphants, we examined the expression of vascular markers. We found that stap2 morphants showed a decreased expression of vascular markers flk and stabilin, consistent with the role of stap2 in vascular development. While loss of stap2 cause vascular defects, we expect overexpression of stap2 will enhance vascular growth. Overexpression of stap2 enhanced the fluorescent signals in vessels and the growth of ISV. These data suggest that stap2 is necessary and sufficient to promote vascular development.We further examine the interaction between stap2 and Vegf signals, we showed stap2 is regulated by VEGF signals to control ISV growth. Together, we conclude that our microarray results identify many vascular genes acting downstream of isl2/coupTFIb pathway. We also showed that stap2 plays an important role for vascular development in zebrafish. Thus, our study provided an useful information for the regulating network in the field of vascular development.
The Dynamic Ontogenetic Structure of Mandibular Morphological Integration
Authors: Holmes Megan A , McNulty Margaret A. , Mussell Jason C. , DeLeon Valerie B. 
Published: November 2023

Mammalian crania demonstrate conserved patterns of morphological integration and simultaneously possess varying magnitudes (degrees) of integration. Thus, it has been posited that plasticity in magnitudes of integration allow for adaptive diversification, while conserved patterns of integration reflect stabilizing selection within the population. This has not been widely tested in the mandible but could have profound implications for mandibular diversity. Here, we test these relationships in the mandible using a mouse model with a mutation that disturbs bone growth in the skull (Fgfr2C342Y). Pattern and magnitude of integration were compared between age‐matched wild‐type and mutant mice. The effect of growth was tested by comparing age groups within each genotype.Micro‐CT scans were collected from wild‐type and mutant mouse skulls at three developmental stages: embryonic, peri‐weaning and adult (n=152). Three‐dimensional landmark coordinate data were collected from each mandible. Procrustes superimposition was applied to coordinate data to produce shape variables. Covariance matrices and scaled variance of eigenvalues were generated from shape variables and used to compare patterns and magnitudes of covariance, respectively. Significance was determined via permutation tests.Patterns of integration in the mandible were conserved between genotype at each developmental stage (R‐obs<0.5; p<0.0001), while magnitude of integration was significantly higher in the mutant mouse model (p<0.001). Interestingly, when pattern was compared among age groups, integration was no longer conserved (R‐obs<0.2; p>0.05). Magnitude of integration decreased markedly with age in the wild‐type mice but increased as mutants aged (p>0.01). The way in which traits covaried (pattern) did not alter despite a perturbation in mandibular growth. The Fgfr2C342Y mutation did however, result in a significant change in magnitude of integration. Importantly, the only instance where pattern of integration altered was between developmental periods. These results support the suggestion that pattern of integration stabilizes shape covariance in a population, and different magnitudes of integration allow for plasticity in shape. Furthermore, both pattern and magnitude change throughout ontogeny, as separate functional and developmental demands occur. Future research should consider the complex influence of pattern, magnitude and development on the diversification of mandibular shape.Support or Funding InformationSupported by NIH #R03DE019816 and Louisiana Board of Regents Traditional Enhancement Grant
microRNA in cardiac arrhythmias
Authors: Wang Jun , Bai Yan , Li Na , Ye Wenduo , Zhang Min , Greene Stephanie B , Tao Ye , Chen Yiping , Wehrens Xander H. T. , Martin James F 
Published: November 2023

microRNAs (miRs) are small endogenous non‐coding RNAs that function in biologic processes primarily via post‐transcriptional gene silencing. Previous genome‐wide association studies and mouse studies indicated homeobox gene Pitx2 as a predisposition gene of atrial fibrillation (AF), the most common sustained cardiac arrhythmia. To identify miRs function in AF, we performed miR expression profiling and ChIP‐sequencing (ChIP‐Seq) using different Pitx2 alleles. We found that Pitx2 positively regulates miR‐17‐92 and its homologous cluster miR‐106b‐25. Both miR‐17‐92 and miR‐106b‐25 deficient mice had pacing‐induced AF under programmed stimulation. Electrocardiogram telemetry further revealed that mice with miR‐17‐92 cardiac‐specific conditional knockout (CKO) developed prolonged PR intervals whereas mice with miR‐17‐92 cardiac‐specific CKO and miR‐106b‐25 heterozygosity developed sinoatrial node dysfunction(SND). Both arrhythmias are AF risk factors in humans. Importantly, miR‐17‐92 and miR‐106b‐25 directly repress the sinoatrial node (SAN)‐specific gene expression such as Shox2 and Tbx3. Together, these findings provided a novel insight for miR function in prevention of AF. In addition, our most recent data indicated that miR‐17‐92 is active in the cardiac conduction system (CCS) and mice with miR‐17‐92 CCS‐specific CKO had cardiac arrhythmias, indicating a miR function in the CCS homeostasis.Support or Funding InformationThis work was supported by a 2012–2013 Michael Bilitch Fellowship in Cardiac Pacing and Electrophysiology from the Heart Rhythm Society (to J.W.), American Heart Association (AHA) Grant 14SDG19840000 (to J.W.), AHA Grant 12PRE11720003 (to Y.B.), AHA Grant 12BGIA12050207 (to N.L.), AHA Grant 13PRE13750003 (to W.Y.), National Institutes of Health (NIH) Grant 1F32HL105041 (to Y.T.), AHA Grant 13EIA14560061, NIH Grants R01‐HL089598 and R01‐HL091947, the Fondation Leducq Alliance for CaMKII Signaling in Heart (X.H.T.W.), NIH Grant 5R01HL118761 (to J.F.M.), NIH Grant R01DE17792 (to Y.C.), and the Vivian L. Smith foundation (J.F.M.).
Activity‐based glycosidase profiling: monitoring glycosphingolipid metabolism in health and disease
Authors: Overkleeft Herman 
Published: November 2023

Activity‐based protein profiling allow the discovery and monitoring of active enzymes in their biological surroundings. An activity‐based probe (ABP) consists of a substrate analogue featuring a reactive group that binds to the enzyme (or enzyme family) of interest in a covalent and irreversible manner, as well as a reporter moiety, which can be an affinity tag, a fluorophore or a bioorthogonal tag. Enzyme families that form a covalent bond in their substrate processing are most suited for ABP design and therefore most ABP applications in the past decade have focused on ABPs for serene hydrolyses, cysteine proteases and threonine proteases. Retaining glycosidases employ a Koshland two‐step double displacement in the hydrolysis of glycosidic bonds. During the catalytic cycle a covalent bond with the substrate glycoside is formed along with expulsion of the aglycon leaving group. The natural product, cyclophellitol, is a potent mechanism‐based retaining beta‐glucosidase inhibitor that capitalises on this mechanism by forming a stable, covalent enzyme‐inhibitor adduct. Grafting a reporter molecule on cyclophellitol or its nitrogen analogue, cyclophellitol aziridine, delivers a potent and selective retaining beta‐glucosidase ABP. Variation of the cyclophellitol/cyclophellitol aziridine configuration yields ABPs selective for a variety of retaining gluycosidases including lysosomal galactocerebrosidase, lysosomal and ER alpha glucosidase and a number of retaining beta‐glucuronidases. In this contribution I will present our work on the design synthesis and application of a broad panel of retaining glycosidase ABPs. I will focus on the application of these tools in the functioning of the underlying glycosidases in health and disease, with special attention to glycosidases involved in lysosomal storage disorders.Support or Funding InformationERC Advanced Grand ‘CHEMBIOSPHING’ Netherlands Organization for Scientific Research
Snapshots Of Benzylsuccinate Synthase: Getting A Handle On Toluene Degradation
Authors: Drennan Catherine L , Funk Michael A 
Published: November 2023

Anaerobic bacteria play a critical role in bioremediation of oil spills and industrial pollution. The first step in the anaerobic degradation of toluene, a major component of gasoline, is the addition of toluene to fumarate to yield benzylsuccinate. The enzyme responsible is benzylsuccinate synthase (BSS), a member of glycyl radical enzyme family, which uses free radical chemistry to break and form C‐H and C‐C bonds. Here, crystallographic snapshots of BSS are presented that allow us to consider how a glycyl radical enzyme can control substrate specificity to afford activation of the otherwise inert toluene molecule.Support or Funding InformationThis work was supported in part by the National Science Foundation Graduate Research Fellowship under Grant No. 0645960 (M.A.F.). C.L.D is a Howard Hughes Medical Institute (HHMI) Investigator.
Semisynthetic Natural Products for Antibacterial Drug Discovery
Authors: Lee Richard 
Published: November 2023

The pressing need for new treatments for multi‐drug resistant bacterial infections including MDR and XDR tuberculosis has led us to re‐explore the use of natural product antibiotics not normally used to treat target infections, especially those with unique mechanisms of action. Being the product of combinatorial evolution natural products have many advantages over purely synthetic antibacterial leads, but also some notable disadvantages often in regard to spectrum and human metabolism. This presentation will go over our efforts to advance under‐explored antibiotic scaffolds with semi‐synthetic approaches to tackle these disadvantages. Using spectinomycin as an example, synthetic modifications have been applied to overcome native bacterial efflux leading to potent leads with better than predicted in vivo efficacy in complex infection models. These observations have led us to examine active uptake and slow‐growing pathogen targeting hypotheses that are hard wired into many natural products, which are very hard to replicate by direct purely synthetic de novo antibacterial drug design.Support or Funding InformationThis work was supported by research grant R01AI090810 by the National Institutes of Health and the American Lebanese Syrian Associated Charities (ALSAC)