03640nas a2200565 4500008004100000022001400041245014000055210006900195260001300264300001200277490000700289520189600296653004202192653003102234653001502265653001902280653002002299653002402319653001902343653003002362653003202392653001502424653001902439653002802458653001002486653001102496653001602507653004402523653001402567653003602581653002702617100002102644700003102665700001502696700001402711700001502725700002202740700001602762700001202778700002302790700001402813700001302827700001902840700002402859700001502883700001402898700001902912700001502931856012802946 2020 eng d a1469-069100aAssociation of a single nucleotide polymorphism in the ubxn6 gene with long-term non-progression phenotype in HIV-positive individuals.0 aAssociation of a single nucleotide polymorphism in the ubxn6 gen c2020 Jan a107-1140 v263 a
OBJECTIVES: The long-term non-progressors (LTNPs) are a heterogeneous group of HIV-positive individuals characterized by their ability to maintain high CD4 T-cell counts and partially control viral replication for years in the absence of antiretroviral therapy. The present study aims to identify host single nucleotide polymorphisms (SNPs) associated with non-progression in a cohort of 352 individuals.
METHODS: DNA microarrays and exome sequencing were used for genotyping about 240 000 functional polymorphisms throughout more than 20 000 human genes. The allele frequencies of 85 LTNPs were compared with a control population. SNPs associated with LTNPs were confirmed in a population of typical progressors. Functional analyses in the affected gene were carried out through knockdown experiments in HeLa-P4, macrophages and dendritic cells.
RESULTS: Several SNPs located within the major histocompatibility complex region previously related to LTNPs were confirmed in this new cohort. The SNP rs1127888 (UBXN6) surpassed the statistical significance of these markers after Bonferroni correction (q = 2.11 × 10). An uncommon allelic frequency of rs1127888 among LTNPs was confirmed by comparison with typical progressors and other publicly available populations. UBXN6 knockdown experiments caused an increase in CAV1 expression and its accumulation in the plasma membrane. In vitro infection of different cell types with HIV-1 replication-competent recombinant viruses caused a reduction of the viral replication capacity compared with their corresponding wild-type cells expressing UBXN6.
CONCLUSIONS: A higher prevalence of Ala31Thr in UBXN6 was found among LTNPs within its N-terminal region, which is crucial for UBXN6/VCP protein complex formation. UBXN6 knockdown affected CAV1 turnover and HIV-1 replication capacity.
10aAdaptor Proteins, Vesicular Transport10aAutophagy-Related Proteins10aCaveolin 110aCohort Studies10aDendritic Cells10aDisease Progression10aGene Frequency10aGene Knockdown Techniques10aGenetic Association Studies10aHeLa Cells10aHIV Infections10aHIV Long-Term Survivors10aHIV-110aHumans10aMacrophages10aOligonucleotide Array Sequence Analysis10aPhenotype10aPolymorphism, Single Nucleotide10awhole exome sequencing1 aDíez-Fuertes, F1 aDe La Torre-Tarazona, H, E1 aCalonge, E1 aPernas, M1 aBermejo, M1 aGarcía-Pérez, J1 aÁlvarez, A1 aCapa, L1 aGarcía-García, F1 aSaumoy, M1 aRiera, M1 aBoland-Auge, A1 aLópez-Galíndez, C1 aLathrop, M1 aDopazo, J1 aSakuntabhai, A1 aAlcamí, J uhttp://clinbioinfosspa.es/content/association-single-nucleotide-polymorphism-ubxn6-gene-long-term-non-progression-phenotype02237nas a2200409 4500008004100000022001400041245006800055210006700123260001300190300001400203490000700217520102100224653001201245653002701257653002701284653001501311653001301326653003001339653000901369653002701378653001201405653002601417653002701443653002601470100001901496700001801515700002001533700002901553700001601582700001501598700002701613700002001640700002001660700002701680700001901707856010101726 2016 eng d a1551-400500aDysfunctional mitochondrial fission impairs cell reprogramming.0 aDysfunctional mitochondrial fission impairs cell reprogramming c2016 Dec a3240-32500 v153 aWe have recently shown that mitochondrial fission is induced early in reprogramming in a Drp1-dependent manner; however, the identity of the factors controlling Drp1 recruitment to mitochondria was unexplored. To investigate this, we used a panel of RNAi targeting factors involved in the regulation of mitochondrial dynamics and we observed that MiD51, Gdap1 and, to a lesser extent, Mff were found to play key roles in this process. Cells derived from Gdap1-null mice were used to further explore the role of this factor in cell reprogramming. Microarray data revealed a prominent down-regulation of cell cycle pathways in Gdap1-null cells early in reprogramming and cell cycle profiling uncovered a G2/M growth arrest in Gdap1-null cells undergoing reprogramming. High-Content analysis showed that this growth arrest was DNA damage-independent. We propose that lack of efficient mitochondrial fission impairs cell reprogramming by interfering with cell cycle progression in a DNA damage-independent manner.
10aAnimals10aCell Cycle Checkpoints10aCellular Reprogramming10aDNA Damage10aG2 Phase10aGene Knockdown Techniques10aMice10aMitochondrial Dynamics10aMitosis10aNerve Tissue Proteins10aPluripotent Stem Cells10aTranscription Factors1 aPrieto, Javier1 aLeón, Marian1 aPonsoda, Xavier1 aGarcia-Garcia, Francisco1 aBort, Roque1 aSerna, Eva1 aBarneo-Muñoz, Manuela1 aPalau, Francesc1 aDopazo, Joaquin1 aLópez-García, Carlos1 aTorres, Josema uhttp://clinbioinfosspa.es/content/dysfunctional-mitochondrial-fission-impairs-cell-reprogramming02926nas a2200445 4500008004100000022001400041245009600055210006900151260000900220300001100229490000600240520152900246653002101775653001401796653002101810653002301831653002101854653001101875653002001886653003001906653004301936653003001979653001102009653001602020653002602036653002402062653002602086100002702112700002102139700002902160700001602189700003002205700002002235700001702255700001802272700002402290700002002314700002102334856012502355 2013 eng d a1932-620300aMammosphere formation in breast carcinoma cell lines depends upon expression of E-cadherin.0 aMammosphere formation in breast carcinoma cell lines depends upo c2013 ae772810 v83 aTumors are heterogeneous at the cellular level where the ability to maintain tumor growth resides in discrete cell populations. Floating sphere-forming assays are broadly used to test stem cell activity in tissues, tumors and cell lines. Spheroids are originated from a small population of cells with stem cell features able to grow in suspension culture and behaving as tumorigenic in mice. We tested the ability of eleven common breast cancer cell lines representing the major breast cancer subtypes to grow as mammospheres, measuring the ability to maintain cell viability upon serial non-adherent passage. Only MCF7, T47D, BT474, MDA-MB-436 and JIMT1 were successfully propagated as long-term mammosphere cultures, measured as the increase in the number of viable cells upon serial non-adherent passages. Other cell lines tested (SKBR3, MDA-MB-231, MDA-MB-468 and MDA-MB-435) formed cell clumps that can be disaggregated mechanically, but cell viability drops dramatically on their second passage. HCC1937 and HCC1569 cells formed typical mammospheres, although they could not be propagated as long-term mammosphere cultures. All the sphere forming lines but MDA-MB-436 express E-cadherin on their surface. Knock down of E-cadherin expression in MCF-7 cells abrogated its ability to grow as mammospheres, while re-expression of E-cadherin in SKBR3 cells allow them to form mammospheres. Therefore, the mammosphere assay is suitable to reveal stem like features in breast cancer cell lines that express E-cadherin.
10aBreast Neoplasms10aCadherins10aCell Line, Tumor10aCell Proliferation10aCluster Analysis10aFemale10agene expression10aGene Expression Profiling10aGene Expression Regulation, Neoplastic10aGene Knockdown Techniques10aHumans10aMCF-7 Cells10aNeoplastic Stem Cells10aSpheroids, Cellular10aTumor Cells, Cultured1 aIglesias, Juan, Manuel1 aBeloqui, Izaskun1 aGarcia-Garcia, Francisco1 aLeis, Olatz1 aVazquez-Martin, Alejandro1 aEguiara, Arrate1 aCufi, Silvia1 aPavon, Andres1 aMenendez, Javier, A1 aDopazo, Joaquin1 aMartin, Angel, G uhttp://clinbioinfosspa.es/content/mammosphere-formation-breast-carcinoma-cell-lines-depends-upon-expression-e-cadherin-003355nas a2200529 4500008004100000022001400041245009200055210007000147260001300217300001100230490000600241520169800247653001801945653001801963653002301981653001502004653002602019653001302045653001702058653003002075653003002105653001702135653001102152653002102163653002702184653005002211653002202261653001202283653001502295653002702310653001502337653004402352653002102396653002402417100002002441700002402461700001802485700002002503700002902523700002002552700002202572700002302594700002302617700003002640700002202670856013302692 2012 eng d a2629-327700aIL1β induces mesenchymal stem cells migration and leucocyte chemotaxis through NF-κB.0 aIL1β induces mesenchymal stem cells migration and leucocyte chem c2012 Sep a905-160 v83 aMesenchymal stem cells are often transplanted into inflammatory environments where they are able to survive and modulate host immune responses through a poorly understood mechanism. In this paper we analyzed the responses of MSC to IL-1β: a representative inflammatory mediator. Microarray analysis of MSC treated with IL-1β revealed that this cytokine activateds a set of genes related to biological processes such as cell survival, cell migration, cell adhesion, chemokine production, induction of angiogenesis and modulation of the immune response. Further more detailed analysis by real-time PCR and functional assays revealed that IL-1β mainly increaseds the production of chemokines such as CCL5, CCL20, CXCL1, CXCL3, CXCL5, CXCL6, CXCL10, CXCL11 and CX(3)CL1, interleukins IL-6, IL-8, IL23A, IL32, Toll-like receptors TLR2, TLR4, CLDN1, metalloproteins MMP1 and MMP3, growth factors CSF2 and TNF-α, together with adhesion molecules ICAM1 and ICAM4. Functional analysis of MSC proliferation, migration and adhesion to extracellular matrix components revealed that IL-1β did not affect proliferation but also served to induce the secretion of trophic factors and adhesion to ECM components such as collagen and laminin. IL-1β treatment enhanced the ability of MSC to recruit monocytes and granulocytes in vitro. Blockade of NF-κβ transcription factor activation with IκB kinase beta (IKKβ) shRNA impaired MSC migration, adhesion and leucocyte recruitment, induced by IL-1β demonstrating that NF-κB pathway is an important downstream regulator of these responses. These findings are relevant to understanding the biological responses of MSC to inflammatory environments.
10aCell Adhesion10aCell Movement10aCell Proliferation10aChemokines10aChemotaxis, Leukocyte10aCollagen10aFibronectins10aGene Expression Profiling10aGene Knockdown Techniques10aHEK293 Cells10aHumans10aI-kappa B Kinase10aInflammation Mediators10aIntercellular Signaling Peptides and Proteins10aInterleukin-1beta10aLaminin10aLeukocytes10aMesenchymal Stem Cells10aNF-kappa B10aOligonucleotide Array Sequence Analysis10aRNA Interference10aSignal Transduction1 aCarrero, Rubén1 aCerrada, Inmaculada1 aLledó, Elisa1 aDopazo, Joaquin1 aGarcia-Garcia, Francisco1 aRubio, Mari-Paz1 aTrigueros, César1 aDorronsoro, Akaitz1 aRuiz-Sauri, Amparo1 aMontero, José, Anastasio1 aSepúlveda, Pilar uhttp://clinbioinfosspa.es/content/il1%CE%B2-induces-mesenchymal-stem-cells-migration-and-leucocyte-chemotaxis-through-nf-%CE%BAb