2022
A genome-scale gain-of-function CRISPR screen in CD8 T cells identifies proline metabolism as a means to enhance CAR-T therapy
Ye L, Park JJ, Peng L, Yang Q, Chow RD, Dong MB, Lam SZ, Guo J, Tang E, Zhang Y, Wang G, Dai X, Du Y, Kim HR, Cao H, Errami Y, Clark P, Bersenev A, Montgomery RR, Chen S. A genome-scale gain-of-function CRISPR screen in CD8 T cells identifies proline metabolism as a means to enhance CAR-T therapy. Cell Metabolism 2022, 34: 595-614.e14. PMID: 35276062, PMCID: PMC8986623, DOI: 10.1016/j.cmet.2022.02.009.Peer-Reviewed Original ResearchConceptsCAR T cellsT cell-based immunotherapyRight molecular targetCell-based immunotherapyCAR-T therapyChimeric antigen receptorMultiple cancer modelsCAR-T efficacyFunction CRISPR screensCD8 TPrimary CD8Immune functionImmunological diseasesImmune boosterCancer modelAntigen receptorDistinct gene expressionMolecular targetsCRISPR activation screensMetabolic programsImmunological analysisTherapyCancerEfficacyActivation screens
2020
Phosphoinositide 3′-Kinase γ Facilitates Polyomavirus Infection
Clark P, Gee GV, Albright BS, Assetta B, Han Y, Atwood WJ, DiMaio D. Phosphoinositide 3′-Kinase γ Facilitates Polyomavirus Infection. Viruses 2020, 12: 1190. PMID: 33092168, PMCID: PMC7589550, DOI: 10.3390/v12101190.Peer-Reviewed Original ResearchConceptsCRISPR/Cas9-mediated knockoutGenetic knockdown approachesCas9-mediated knockoutDNA tumor virusesProgressive multifocal leukoencephalopathyInfection defectPolyomavirus infectionCellular proteinsKnockdown approachSVG-A cellsPolyomavirus entryMolecular eventsVirus internalizationTumor virusPI3Kγ pathwayPIK3R5PI3KγGlial cellsKnockoutMultifocal leukoencephalopathyImmunosuppressed peopleCellsHuman polyomavirusesJC polyomavirusPolyomavirus
2016
The Receptor for Advanced Glycation Endproducts Drives T Cell Survival and Inflammation in Type 1 Diabetes Mellitus
Durning SP, Preston-Hurlburt P, Clark PR, Xu D, Herold KC, Group T. The Receptor for Advanced Glycation Endproducts Drives T Cell Survival and Inflammation in Type 1 Diabetes Mellitus. The Journal Of Immunology 2016, 197: 3076-3085. PMID: 27655844, PMCID: PMC5101164, DOI: 10.4049/jimmunol.1600197.Peer-Reviewed Original ResearchMeSH KeywordsAdolescentAdultAsymptomatic DiseasesCD8-Positive T-LymphocytesCell SurvivalCells, CulturedChildDiabetes Mellitus, Type 1Disease ProgressionFemaleGene Expression ProfilingHumansImmunologic MemoryInflammationLymphocyte ActivationMaleReceptor for Advanced Glycation End ProductsRiskSignal TransductionUp-RegulationYoung AdultConceptsDamage-associated molecular patternsT cellsRAGE expressionT1D patientsInflammatory functionsRisk relativesCell activationHigh mobility group box 1Mobility group box 1Advanced glycated endproductsChronic autoimmune responseMolecular patternsEffector memory cellsHealthy control subjectsExpression of RAGEGroup box 1Type 1 diabetesAdvanced glycation endproductsT cell survivalAutoimmune responseAutoimmune diseasesControl subjectsDisease onsetRisk subjectsCell injury
2015
Tumor Necrosis Factor Disrupts Claudin-5 Endothelial Tight Junction Barriers in Two Distinct NF-κB-Dependent Phases
Clark PR, Kim RK, Pober JS, Kluger MS. Tumor Necrosis Factor Disrupts Claudin-5 Endothelial Tight Junction Barriers in Two Distinct NF-κB-Dependent Phases. PLOS ONE 2015, 10: e0120075. PMID: 25816133, PMCID: PMC4376850, DOI: 10.1371/journal.pone.0120075.Peer-Reviewed Original ResearchCell Membrane PermeabilityCells, CulturedClaudin-5DermisEndothelium, VascularHuman Umbilical Vein Endothelial CellsHumansMicroscopy, FluorescenceMyosin Light ChainsMyosin-Light-Chain KinaseNF-kappa BPhosphorylationRho-Associated KinasesRNA, Small InterferingSignal TransductionTight JunctionsTumor Necrosis Factor-alpha
2011
MEK5 is Activated by Shear Stress, Activates ERK5 and Induces KLF4 to Modulate TNF Responses in Human Dermal Microvascular Endothelial Cells
CLARK PR, JENSEN TJ, KLUGER MS, MORELOCK M, HANIDU A, QI Z, TATAKE RJ, POBER JS. MEK5 is Activated by Shear Stress, Activates ERK5 and Induces KLF4 to Modulate TNF Responses in Human Dermal Microvascular Endothelial Cells. Microcirculation 2011, 18: 102-117. PMID: 21166929, PMCID: PMC3075844, DOI: 10.1111/j.1549-8719.2010.00071.x.Peer-Reviewed Original ResearchMeSH KeywordsBase SequenceCells, CulturedEndothelial CellsEnzyme ActivationGene Expression ProfilingGene Knockdown TechniquesHumansKruppel-Like Factor 4Kruppel-Like Transcription FactorsMAP Kinase Kinase 5Mitogen-Activated Protein Kinase 7Oligonucleotide Array Sequence AnalysisRNA, MessengerRNA, Small InterferingShear StrengthStress, MechanicalTumor Necrosis Factor-alphaConceptsInduction of KLF4Inflammatory responseKLF4 expressionTNF responseInhibits Inflammatory ResponsesPro-inflammatory changesAnti-inflammatory pathwayICAM-1 expressionHuman dermal microvascular endothelial cellsDermal microvascular endothelial cellsMicrovascular endothelial cellsMicrovascular ECsWestern blottingEndothelial cellsRole of ERK5Protein expressionQRT-PCRSiRNA knockdownERK5 phosphorylationHDMECPrincipal siteHuman skinERK5 activationImmunofluorescence microscopyInduction
2007
Knockdown of TNFR1 by the sense strand of an ICAM-1 siRNA: dissection of an off-target effect
Clark PR, Pober JS, Kluger MS. Knockdown of TNFR1 by the sense strand of an ICAM-1 siRNA: dissection of an off-target effect. Nucleic Acids Research 2007, 36: 1081-1097. PMID: 18096615, PMCID: PMC2275081, DOI: 10.1093/nar/gkm630.Peer-Reviewed Original ResearchConceptsTumor necrosis factor receptor 1Endothelial cellsIntercellular adhesion molecule-1ICAM-1 siRNAAnti-TNF activityCultured human umbilical vein endothelial cellsICAM-1 expressionAdhesion molecule-1Off-target effectsNecrosis factor receptor 1Human umbilical vein ECHuman umbilical vein endothelial cellsFactor receptor 1Umbilical vein endothelial cellsExpression of mRNAIL-1 activationVein endothelial cellsLocal inflammationTNFR1 mRNANecrosis factorEarly TNFICAM-1VCAM-1E-selectinTransfection of ECs
2006
Increased ICAM-1 Expression Causes Endothelial Cell Leakiness, Cytoskeletal Reorganization and Junctional Alterations
Clark PR, Manes TD, Pober JS, Kluger MS. Increased ICAM-1 Expression Causes Endothelial Cell Leakiness, Cytoskeletal Reorganization and Junctional Alterations. Journal Of Investigative Dermatology 2006, 127: 762-774. PMID: 17195014, DOI: 10.1038/sj.jid.5700670.Peer-Reviewed Original ResearchConceptsEC barrier functionCell junctionsHuman dermal microvascular ECsSmall-interfering RNA knockdownEC shape changesEndothelial cellsG actin ratioActin cytoskeletonShape changesCytoskeleton organizationCell elongationICAM-1 inductionCytoskeletal reorganizationIntracellular regionCell shapeRNA knockdownHuman dermal microvascular endothelial cellsDermal microvascular endothelial cellsZona occludens-1NF-kappaB activationJunctional alterationsCytoskeletonBarrier functionCell leakinessMicrovascular endothelial cells