DOCSLIB.ORG

Transcriptional Profile of Human Anti-Inflammatory Macrophages Under Homeostatic, Activating and Pathological Conditions

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Transcriptional Profile of Human Anti-Inflammatory Macrophages Under Homeostatic, Activating and Pathological Conditions Universidad Complutense de Madrid Facultad de Ciencias Químicas Dpto. de Bioquímica y Biología Molecular I TRANSCRIPTIONAL PROFILE OF HUMAN ANTI-INFLAMMATORY MACROPHAGES UNDER HOMEOSTATIC, ACTIVATING AND PATHOLOGICAL CONDITIONS Perfil transcripcional de macrófagos antiinflamatorios humanos en condiciones de homeostasis, activación y patológicas. Víctor Delgado Cuevas Tesis Doctoral Madrid 2016 Universidad Complutense de Madrid Facultad de Ciencias Químicas Dpto. de Bioquímica y Biología Molecular I TRANSCRIPTIONAL PROFILE OF HUMAN ANTI-INFLAMMATORY MACROPHAGES UNDER HOMEOSTATIC, ACTIVATING AND PATHOLOGICAL CONDITIONS Perfil transcripcional de macrófagos antiinflamatorios humanos en condiciones de homeostasis, activación y patológicas. Este trabajo ha sido realizado por Víctor Delgado Cuevas para optar al grado de Doctor en el Centro de Investigaciones Biológicas de Madrid (CSIC), bajo la dirección de la Dra. María Marta Escribese Alonso y el Dr. Ángel Luís Corbí López Fdo. Dra. María Marta Escribese Alonso Fdo. Dr. Ángel Luís Corbí López Gracias a los que me enseñan, me pagan, me ayudan, me prestan, me acompañan, me soportan, me quieren, me dan de comer, me invitan a beber y me llevan a bailar. Esto no ha hecho más que empezar. INDEX ABBREVIATIONS 9 ABSTRACT 13 RESUMEN 15 INTRODUCTION 19 MACROPHAGE ONTOGENY 21 MACROPHAGES IN INFLAMMATION 21 TISSUE-RESIDENT MACROPHAGES 23 MACROPHAGE ACTIVATION 26 Toll-like receptors 28 MAFB 31 Structural features and expression 31 MAFB functions 32 MAFB-related pathologies 34 Multicentric carpotarsal osteolysis (MCTO) 35 OBJECTIVES 37 RESULTS 39 CHAPTER ONE: A novel set of genes that define the activation state of human anti-inflammatory macrophages 39 Differential cytokine LPS responsiveness of GM-MØ and M-MØ 41 Differential LPS-initiated intracellular signaling pathways in GM-MØ and M-MØ 45 Definition of the LPS-regulated transcriptional signature of GM-MØ and M-MØ 47 Biological function of the genes upregulated by LPS exclusively in anti-inflammatory M-MØ 50 Contribution of ERK, JNK and p38 activation to the M-MØ-specific LPS-dependent transcriptional profile 50 Physiologic and pathologic relevance of the M-MØ-specific LPS-induced transcriptional profile 54 Table III. LPS-activated GM-MØ and M-MØ: microarray results. Digital format. See attached CD on the reverse of the back cover. Table IV. Genes regulated by LPS exclusively in GM-MØ or M-MØ 56 Table V. Genes regulated by LPS in both GM-MØ and M-MØ 60 Table VI. Effect of MAPK inhibitors on the LPS-induced transcriptional profile of M-MØ 63 CHAPTER TWO: MAFB determines human macrophage anti-inflammatory polarization: pathological relevance for Multicentric carpotarsal osteolysis 65 MAFB expression in human macrophages under homeostatic and anti-inflammatory conditions. 67 MAFB controls the acquisition of the anti-inflammatory transcriptional profile of M-MØ. 67 The M-MØ-specific macrophage transcriptome is altered in macrophages derived from Multicentric carpotarsal osteolysis monocytes. 74 MAFB also influences the LPS responsiveness of human macrophages. 78 Co-expression of MAFB and MAFB-regulated genes in human macrophages in vivo. 80 Table VII. Probes and annotated genes with altered expression in siMAFB M-MØ. 82 Table VIII. Probes and annotated genes with altered expression in MCTO#1 M-MØ 89 DISCUSSION 99 Human M-MØ activation 101 MAFB-directed human macrophage anti-inflammatory polarization 105 Pathological relevance of MAFB-driven macrophage polarization 108 CONCLUSSIONS 113 EXPERIMENTAL PROCEDURES 117 REFERENCES 125 ABBREVIATIONS AHR: aryl hydrocarbon receptor DMSO: dimethyl sulfoxide ARNT: aryl hydrocarbon receptor nuclear EGF: epidermal growth factor translocator ELISA: enzyme-linked immunosorbent assay bHLH-PAS: basic helix-loop-helix-PER-ARNT- SIM Enrichr: interactive and collaborative HTML5 gene list enrichment tool BMP: bone morphogenic protein ERK: extracellular signal-regulated kinase CCL: C-C motif chemokine ligand FDR: false discovery rate CCR: C-C chemokyne receptor FOS: Fos proto-oncogene, AP-1 transcription CD: cluster of differentiation factor subunit ChIP-seq: chromatin immunoprecipitation GATA6: GATA binding protein 6 sequencing GM-CSF: granulocyte macrophage-colony ChIP: chromatin immunoprecipitation assay stimulating factor CLEC: C-type lectin GM-MØ: GM-CSF-polarized macrophage CRE: cAMP-responsive element GSEA: gene set enrichment analysis CREB: cAMP response element binding protein GSK3: glycogen synthase kinase 3 CSF1R: colony-stimulating factor 1 receptor HIF: hypoxia-inducible factor CXCL: C-X-C motif ligand HMGB1: high mobility group box 1 protein DAMP: damage-associated molecular pattern HTR: 5-hydroxytryptamine receptor 11 IFN: interferon pMAFB: MAFB pCDNA3.1(+) expression vector IL: interleukin PPAR: peroxisome proliferator activated receptor IL-34-MØ: IL-34-polarized macrophage PRR: pattern recognition receptor IRAK: interleukin-1 receptor-associated kinase qRT-PCR: quantitative real-time polymerase chain reaction IRF: interferon-regulatory transcription factor RANKL: receptor activator of nuclear factor- JNK: JUN N-terminal kinase kappaB ligand LPS: lipopolysaccharide RPMI: Roswell Park Memorial Institute medium LRR: leucine-rich repeat SARM: sterile alpha and TIR motif-containing protein LXR: liver X receptor SIM: single-minded family bHLH transcription M-CSF: macrophage-colony stimulating factor factor 1 M-MØ: M-CSF-polarized macrophage siRNA: small interfering RNA MAF: v-Maf avian musculoaponeurotic SLC40A1: solute carrier family 40 member 1 fibrosarcoma oncogene homolog SOCS: suppressor of cytokine signaling MAL: MYD88-adapter like SPIC: Spi-C transcription factor MAPK: mitogen-activated protein kinase STAT: signal transducer and activator of MARE: Maf-recognition element transcription MCTO: multicentric carpotarsal osteolysis TCDD: 2,3,7,8-tetrachlorodibenzo-p-dioxin MEK: MAPK/ERK kinase TGF: transforming growth factor MHC: major histocompatibility complex TIR: Toll/interleukin-1 receptor MITF: melanogenesis associated transcription TLR: toll-like receptor factor TNF: tumor necrosis factor MMP: matrix metalloproteinase TRAF: TNF receptor-associated factor MONA: multicentric osteolysis, nodulosis and arthropathy TRAM: TRIF-related adaptor molecule MYD88: myeloid differentiation primary response TRAP: tartrate-resistant acid phosphatase 88 TRE: 12-O-tetradecanoyl phorbol 13-acetate NFATc1: nuclear factor of activated T cell 1 (TPA)-responsive element k NF B: nuclear factor-kappa B TRIF: TIR domain-containing adaptor protein inducing interferon beta NPAS4: neuronal PAS domain protein 4 VEGF: vascular endothelial growth factor NRL: neural retina leucine zipper protein WT: wild type p38: p38 mitogen activated protein kinase ZXDC: ZXD family zinc finger C protein PAM3CSK4: N-palmitoyl-S-[2,3-bis (palmitoyloxy)-(2RS)-propyl]-[R]-cysteinyl-[S]- seryl-[S]-lysyl-[S]-lysyl-[S]-lysyl-[S]-lysine PAMP: pathogen-associated molecular pattern PBMC: peripheral blood mononuclear cells PDGF: platelet-derived growth factor 12 ABSTRACT Macrophages are cells of the innate immune system that exhibit a huge phenotypic and functional heterogeneity, what gives macrophages the ability to sense and respond accurately to the needs of their microenvironment. Thus, macrophage differentiation and functions are dependent on the integration of cues provided by their ontogeny, surrounding tissue, microbiota, metabolism and pathogens. In line with their functional plasticity, macrophages are able to both initiate and resolve inflammation. Besides, macrophages have essential roles in development, homeostasis and inflammation and, consequently, deregulation of macrophage functions leads to the onset and maintenance of numerous chronic inflammatory pathologies. Therefore, determination of the molecular basis of the macrophage functional heterogeneity should pave the way for the development of tissue-specific anti-inflammatory therapies. Macrophage activation relies on the recognition of pathogen- and danger-associated molecular patterns from exogenous and endogenous factors by a variety of receptors, like TLRs. Depending on the nature of previous cues received by macrophages, the response to activating stimuli may be pro- or anti- inflammatory. The former is characteristic of macrophages exposed to GM-CSF or IFNγ and results in the production of TNF-α, IL-12, IL-6 and IL-23. The latter is specific for macrophages exposed to M-CSF, IL-4 or glucocorticoids and leads to the secretion of IL-10. Unlike the macrophage pro-inflammatory response, which has been extensively studied, scarce knowledge is currently available on the activation of anti-inflammatory macrophages, especially in humans. To address this issue, we have thoroughly analyzed the transcriptome of TLR-activated, human M-CSF-polarized anti-inflammatory macrophages (M-MØ). Microarray analysis of M-MØ after a short exposure to LPS revealed the existence of a gene set that includes chemokines (e.g., CCL19), signaling molecules (e.g., SOCS2) and growth factors (e.g., BMP6, PDGFA), that is exclusively induced in M-MØ in vitro and whose expression is dependent on the activation of the MAPKs ERK, p38 and JNK. Representative examples of this gene set, including CCL19, 15 SOCS2, BMP6 and PDGFA, are co-expressed in humans in gut and skin-resident macrophages and tumor-associated macrophages in nevi and melanoma. Thus, we have identified a novel gene set whose expression is specific for human activated homeostatic/anti-inflammatory macrophages. This finding sheds light on the mechanisms
Load more

Recommended publications
  • PARSANA-DISSERTATION-2020.Pdf
    DECIPHERING TRANSCRIPTIONAL PATTERNS OF GENE REGULATION: A COMPUTATIONAL APPROACH by Princy Parsana A dissertation submitted to The Johns Hopkins University in conformity with the requirements for the degree of Doctor of Philosophy Baltimore, Maryland July, 2020 © 2020 Princy Parsana All rights reserved Abstract With rapid advancements in sequencing technology, we now have the ability to sequence the entire human genome, and to quantify expression of tens of thousands of genes from hundreds of individuals. This provides an extraordinary opportunity to learn phenotype relevant genomic patterns that can improve our understanding of molecular and cellular processes underlying a trait. The high dimensional nature of genomic data presents a range of computational and statistical challenges. This dissertation presents a compilation of projects that were driven by the motivation to efficiently capture gene regulatory patterns in the human transcriptome, while addressing statistical and computational challenges that accompany this data. We attempt to address two major difficulties in this domain: a) artifacts and noise in transcriptomic data, andb) limited statistical power. First, we present our work on investigating the effect of artifactual variation in gene expression data and its impact on trans-eQTL discovery. Here we performed an in-depth analysis of diverse pre-recorded covariates and latent confounders to understand their contribution to heterogeneity in gene expression measurements. Next, we discovered 673 trans-eQTLs across 16 human tissues using v6 data from the Genotype Tissue Expression (GTEx) project. Finally, we characterized two trait-associated trans-eQTLs; one in Skeletal Muscle and another in Thyroid. Second, we present a principal component based residualization method to correct gene expression measurements prior to reconstruction of co-expression networks.
    [Show full text]
  • Small Cell Ovarian Carcinoma: Genomic Stability and Responsiveness to Therapeutics
    Gamwell et al. Orphanet Journal of Rare Diseases 2013, 8:33 http://www.ojrd.com/content/8/1/33 RESEARCH Open Access Small cell ovarian carcinoma: genomic stability and responsiveness to therapeutics Lisa F Gamwell1,2, Karen Gambaro3, Maria Merziotis2, Colleen Crane2, Suzanna L Arcand4, Valerie Bourada1,2, Christopher Davis2, Jeremy A Squire6, David G Huntsman7,8, Patricia N Tonin3,4,5 and Barbara C Vanderhyden1,2* Abstract Background: The biology of small cell ovarian carcinoma of the hypercalcemic type (SCCOHT), which is a rare and aggressive form of ovarian cancer, is poorly understood. Tumourigenicity, in vitro growth characteristics, genetic and genomic anomalies, and sensitivity to standard and novel chemotherapeutic treatments were investigated in the unique SCCOHT cell line, BIN-67, to provide further insight in the biology of this rare type of ovarian cancer. Method: The tumourigenic potential of BIN-67 cells was determined and the tumours formed in a xenograft model was compared to human SCCOHT. DNA sequencing, spectral karyotyping and high density SNP array analysis was performed. The sensitivity of the BIN-67 cells to standard chemotherapeutic agents and to vesicular stomatitis virus (VSV) and the JX-594 vaccinia virus was tested. Results: BIN-67 cells were capable of forming spheroids in hanging drop cultures. When xenografted into immunodeficient mice, BIN-67 cells developed into tumours that reflected the hypercalcemia and histology of human SCCOHT, notably intense expression of WT-1 and vimentin, and lack of expression of inhibin. Somatic mutations in TP53 and the most common activating mutations in KRAS and BRAF were not found in BIN-67 cells by DNA sequencing.
    [Show full text]
  • Transcriptomic Profiling of Equine and Viral Genes in Peripheral Blood
    pathogens Article Transcriptomic Profiling of Equine and Viral Genes in Peripheral Blood Mononuclear Cells in Horses during Equine Herpesvirus 1 Infection Lila M. Zarski 1, Patty Sue D. Weber 2, Yao Lee 1 and Gisela Soboll Hussey 1,* 1 Department of Pathobiology and Diagnostic Investigation, Michigan State University, East Lansing, MI 48824, USA; [email protected] (L.M.Z.); [email protected] (Y.L.) 2 Department of Large Animal Clinical Sciences, Michigan State University, East Lansing, MI 48824, USA; [email protected] * Correspondence: [email protected] Abstract: Equine herpesvirus 1 (EHV-1) affects horses worldwide and causes respiratory dis- ease, abortions, and equine herpesvirus myeloencephalopathy (EHM). Following infection, a cell- associated viremia is established in the peripheral blood mononuclear cells (PBMCs). This viremia is essential for transport of EHV-1 to secondary infection sites where subsequent immunopathol- ogy results in diseases such as abortion or EHM. Because of the central role of PBMCs in EHV-1 pathogenesis, our goal was to establish a gene expression analysis of host and equine herpesvirus genes during EHV-1 viremia using RNA sequencing. When comparing transcriptomes of PBMCs during peak viremia to those prior to EHV-1 infection, we found 51 differentially expressed equine genes (48 upregulated and 3 downregulated). After gene ontology analysis, processes such as the interferon defense response, response to chemokines, the complement protein activation cascade, cell adhesion, and coagulation were overrepresented during viremia. Additionally, transcripts for EHV-1, EHV-2, and EHV-5 were identified in pre- and post-EHV-1-infection samples. Looking at Citation: Zarski, L.M.; Weber, P.S.D.; micro RNAs (miRNAs), 278 known equine miRNAs and 855 potentially novel equine miRNAs were Lee, Y.; Soboll Hussey, G.
    [Show full text]
  • Protein Interaction Network of Alternatively Spliced Isoforms from Brain Links Genetic Risk Factors for Autism
    ARTICLE Received 24 Aug 2013 | Accepted 14 Mar 2014 | Published 11 Apr 2014 DOI: 10.1038/ncomms4650 OPEN Protein interaction network of alternatively spliced isoforms from brain links genetic risk factors for autism Roser Corominas1,*, Xinping Yang2,3,*, Guan Ning Lin1,*, Shuli Kang1,*, Yun Shen2,3, Lila Ghamsari2,3,w, Martin Broly2,3, Maria Rodriguez2,3, Stanley Tam2,3, Shelly A. Trigg2,3,w, Changyu Fan2,3, Song Yi2,3, Murat Tasan4, Irma Lemmens5, Xingyan Kuang6, Nan Zhao6, Dheeraj Malhotra7, Jacob J. Michaelson7,w, Vladimir Vacic8, Michael A. Calderwood2,3, Frederick P. Roth2,3,4, Jan Tavernier5, Steve Horvath9, Kourosh Salehi-Ashtiani2,3,w, Dmitry Korkin6, Jonathan Sebat7, David E. Hill2,3, Tong Hao2,3, Marc Vidal2,3 & Lilia M. Iakoucheva1 Increased risk for autism spectrum disorders (ASD) is attributed to hundreds of genetic loci. The convergence of ASD variants have been investigated using various approaches, including protein interactions extracted from the published literature. However, these datasets are frequently incomplete, carry biases and are limited to interactions of a single splicing isoform, which may not be expressed in the disease-relevant tissue. Here we introduce a new interactome mapping approach by experimentally identifying interactions between brain-expressed alternatively spliced variants of ASD risk factors. The Autism Spliceform Interaction Network reveals that almost half of the detected interactions and about 30% of the newly identified interacting partners represent contribution from splicing variants, emphasizing the importance of isoform networks. Isoform interactions greatly contribute to establishing direct physical connections between proteins from the de novo autism CNVs. Our findings demonstrate the critical role of spliceform networks for translating genetic knowledge into a better understanding of human diseases.
    [Show full text]
  • CD29 Identifies IFN-Γ–Producing Human CD8+ T Cells With
    + CD29 identifies IFN-γ–producing human CD8 T cells with an increased cytotoxic potential Benoît P. Nicoleta,b, Aurélie Guislaina,b, Floris P. J. van Alphenc, Raquel Gomez-Eerlandd, Ton N. M. Schumacherd, Maartje van den Biggelaarc,e, and Monika C. Wolkersa,b,1 aDepartment of Hematopoiesis, Sanquin Research, 1066 CX Amsterdam, The Netherlands; bLandsteiner Laboratory, Oncode Institute, Amsterdam University Medical Center, University of Amsterdam, 1105 AZ Amsterdam, The Netherlands; cDepartment of Research Facilities, Sanquin Research, 1066 CX Amsterdam, The Netherlands; dDivision of Molecular Oncology and Immunology, Oncode Institute, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands; and eDepartment of Molecular and Cellular Haemostasis, Sanquin Research, 1066 CX Amsterdam, The Netherlands Edited by Anjana Rao, La Jolla Institute for Allergy and Immunology, La Jolla, CA, and approved February 12, 2020 (received for review August 12, 2019) Cytotoxic CD8+ T cells can effectively kill target cells by producing therefore developed a protocol that allowed for efficient iso- cytokines, chemokines, and granzymes. Expression of these effector lation of RNA and protein from fluorescence-activated cell molecules is however highly divergent, and tools that identify and sorting (FACS)-sorted fixed T cells after intracellular cytokine + preselect CD8 T cells with a cytotoxic expression profile are lacking. staining. With this top-down approach, we performed an un- + Human CD8 T cells can be divided into IFN-γ– and IL-2–producing biased RNA-sequencing (RNA-seq) and mass spectrometry cells. Unbiased transcriptomics and proteomics analysis on cytokine- γ– – + + (MS) analyses on IFN- and IL-2 producing primary human producing fixed CD8 T cells revealed that IL-2 cells produce helper + + + CD8 Tcells.
    [Show full text]
  • Supplementary Data
    SUPPLEMENTARY DATA A cyclin D1-dependent transcriptional program predicts clinical outcome in mantle cell lymphoma Santiago Demajo et al. 1 SUPPLEMENTARY DATA INDEX Supplementary Methods p. 3 Supplementary References p. 8 Supplementary Tables (S1 to S5) p. 9 Supplementary Figures (S1 to S15) p. 17 2 SUPPLEMENTARY METHODS Western blot, immunoprecipitation, and qRT-PCR Western blot (WB) analysis was performed as previously described (1), using cyclin D1 (Santa Cruz Biotechnology, sc-753, RRID:AB_2070433) and tubulin (Sigma-Aldrich, T5168, RRID:AB_477579) antibodies. Co-immunoprecipitation assays were performed as described before (2), using cyclin D1 antibody (Santa Cruz Biotechnology, sc-8396, RRID:AB_627344) or control IgG (Santa Cruz Biotechnology, sc-2025, RRID:AB_737182) followed by protein G- magnetic beads (Invitrogen) incubation and elution with Glycine 100mM pH=2.5. Co-IP experiments were performed within five weeks after cell thawing. Cyclin D1 (Santa Cruz Biotechnology, sc-753), E2F4 (Bethyl, A302-134A, RRID:AB_1720353), FOXM1 (Santa Cruz Biotechnology, sc-502, RRID:AB_631523), and CBP (Santa Cruz Biotechnology, sc-7300, RRID:AB_626817) antibodies were used for WB detection. In figure 1A and supplementary figure S2A, the same blot was probed with cyclin D1 and tubulin antibodies by cutting the membrane. In figure 2H, cyclin D1 and CBP blots correspond to the same membrane while E2F4 and FOXM1 blots correspond to an independent membrane. Image acquisition was performed with ImageQuant LAS 4000 mini (GE Healthcare). Image processing and quantification were performed with Multi Gauge software (Fujifilm). For qRT-PCR analysis, cDNA was generated from 1 µg RNA with qScript cDNA Synthesis kit (Quantabio). qRT–PCR reaction was performed using SYBR green (Roche).
    [Show full text]
  • Transposable Elements Resistant to Epigenetic Resetting in the Human Germline Are Epigenetic Hotspots for Development and Disease
    bioRxiv preprint doi: https://doi.org/10.1101/2020.03.19.998930; this version posted March 20, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 1 Transposable elements resistant to epigenetic resetting in the human germline are epigenetic hotspots for development and disease Sabine Dietmann1,2,3,11*, Michael J Keogh4,5,11, Walfred Tang2, Erna Magnusdottir6, Toshihiro Kobayashi7,8, Patrick F Chinnery5,9 and M. Azim Surani 1,2,10* ** 1. Wellcome Trust – Medical Research Council Stem Cell Institute, University of Cambridge, UK 2. Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge, UK 3. Department of Developmental Biology and Division of Nephrology, Washington University School of Medicine, St. Louis, MO, USA 4. Institute of Genetic Medicine, Newcastle University, Newcastle Upon Tyne, UK 5. MRC-Mitochondrial Biology Unit, University of Cambridge, Cambridge, UK 6. Department of Biomedical Science and Anatomy, Faculty of Medicine, University of Iceland, Reykjavik, Iceland 7. Section of Mammalian Transgenesis, Center for Genetic Analysis of Behavior, National Institute for Physiological Sciences, Okazaki, Japan 8. Department of Physiological Sciences, The Graduate University of Advanced Studies, Okazaki, Japan 9. Department of Clinical Neurosciences, Cambridge Biomedical Campus, University of Cambridge, Cambridge, UK 10. Department of Physiology and Development of Neuroscience, University of Cambridge, UK 11. These authors contributed equally * Co-corresponding authors; [email protected]; [email protected] ** lead contact:[email protected] bioRxiv preprint doi: https://doi.org/10.1101/2020.03.19.998930; this version posted March 20, 2020.
    [Show full text]
  • Rab27a Co-Ordinates Actin-Dependent Transport by Controlling Organelle-Associated Motors and Track Assembly Proteins
    ARTICLE https://doi.org/10.1038/s41467-020-17212-6 OPEN Rab27a co-ordinates actin-dependent transport by controlling organelle-associated motors and track assembly proteins Noura Alzahofi1,10, Tobias Welz2,10, Christopher L. Robinson1, Emma L. Page1, Deborah A. Briggs1, Amy K. Stainthorp 1, James Reekes1, David A. Elbe1, Felix Straub2, Wouter W. Kallemeijn 3, Edward W. Tate 3, Philip S. Goff 4, Elena V. Sviderskaya 4, Marta Cantero5,6, Lluis Montoliu 5,6, ✉ Francois Nedelec7, Amanda K. Miles 8, Maryse Bailly 9, Eugen Kerkhoff2 & Alistair N. Hume 1 1234567890():,; Cell biologists generally consider that microtubules and actin play complementary roles in long- and short-distance transport in animal cells. On the contrary, using melanosomes of melanocytes as a model, we recently discovered that the motor protein myosin-Va works with dynamic actin tracks to drive long-range organelle dispersion in opposition to micro- tubules. This suggests that in animals, as in yeast and plants, myosin/actin can drive long- range transport. Here, we show that the SPIRE-type actin nucleators (predominantly SPIRE1) are Rab27a effectors that co-operate with formin-1 to generate actin tracks required for myosin-Va-dependent transport in melanocytes. Thus, in addition to melanophilin/myosin- Va, Rab27a can recruit SPIREs to melanosomes, thereby integrating motor and track assembly activity at the organelle membrane. Based on this, we suggest a model in which organelles and force generators (motors and track assemblers) are linked, forming an organelle-based, cell-wide network that allows their collective activity to rapidly disperse the population of organelles long-distance throughout the cytoplasm.
    [Show full text]
  • Supplementary Materials
    Supplementary materials Supplementary Table S1: MGNC compound library Ingredien Molecule Caco- Mol ID MW AlogP OB (%) BBB DL FASA- HL t Name Name 2 shengdi MOL012254 campesterol 400.8 7.63 37.58 1.34 0.98 0.7 0.21 20.2 shengdi MOL000519 coniferin 314.4 3.16 31.11 0.42 -0.2 0.3 0.27 74.6 beta- shengdi MOL000359 414.8 8.08 36.91 1.32 0.99 0.8 0.23 20.2 sitosterol pachymic shengdi MOL000289 528.9 6.54 33.63 0.1 -0.6 0.8 0 9.27 acid Poricoic acid shengdi MOL000291 484.7 5.64 30.52 -0.08 -0.9 0.8 0 8.67 B Chrysanthem shengdi MOL004492 585 8.24 38.72 0.51 -1 0.6 0.3 17.5 axanthin 20- shengdi MOL011455 Hexadecano 418.6 1.91 32.7 -0.24 -0.4 0.7 0.29 104 ylingenol huanglian MOL001454 berberine 336.4 3.45 36.86 1.24 0.57 0.8 0.19 6.57 huanglian MOL013352 Obacunone 454.6 2.68 43.29 0.01 -0.4 0.8 0.31 -13 huanglian MOL002894 berberrubine 322.4 3.2 35.74 1.07 0.17 0.7 0.24 6.46 huanglian MOL002897 epiberberine 336.4 3.45 43.09 1.17 0.4 0.8 0.19 6.1 huanglian MOL002903 (R)-Canadine 339.4 3.4 55.37 1.04 0.57 0.8 0.2 6.41 huanglian MOL002904 Berlambine 351.4 2.49 36.68 0.97 0.17 0.8 0.28 7.33 Corchorosid huanglian MOL002907 404.6 1.34 105 -0.91 -1.3 0.8 0.29 6.68 e A_qt Magnogrand huanglian MOL000622 266.4 1.18 63.71 0.02 -0.2 0.2 0.3 3.17 iolide huanglian MOL000762 Palmidin A 510.5 4.52 35.36 -0.38 -1.5 0.7 0.39 33.2 huanglian MOL000785 palmatine 352.4 3.65 64.6 1.33 0.37 0.7 0.13 2.25 huanglian MOL000098 quercetin 302.3 1.5 46.43 0.05 -0.8 0.3 0.38 14.4 huanglian MOL001458 coptisine 320.3 3.25 30.67 1.21 0.32 0.9 0.26 9.33 huanglian MOL002668 Worenine
    [Show full text]
  • Exploring the Relationship Between Gut Microbiota and Major Depressive Disorders
    E3S Web of Conferences 271, 03055 (2021) https://doi.org/10.1051/e3sconf/202127103055 ICEPE 2021 Exploring the Relationship between Gut Microbiota and Major Depressive Disorders Catherine Tian1 1Shanghai American School, Shanghai, China Abstract. Major Depressive Disorder (MDD) is a psychiatric disorder accompanied with a high rate of suicide, morbidity and mortality. With the symptom of an increasing or decreasing appetite, there is a possibility that MDD may have certain connections with gut microbiota, the colonies of microbes which reside in the human digestive system. In recent years, more and more studies started to demonstrate the links between MDD and gut microbiota from animal disease models and human metabolism studies. However, this relationship is still largely understudied, but it is very innovative since functional dissection of this relationship would furnish a new train of thought for more effective treatment of MDD. In this study, by using multiple genetic analytic tools including Allen Brain Atlas, genetic function analytical tools, and MicrobiomeAnalyst, I explored the genes that shows both expression in the brain and the digestive system to affirm that there is a connection between gut microbiota and the MDD. My approach finally identified 7 MDD genes likely to be associated with gut microbiota, implicating 3 molecular pathways: (1) Wnt Signaling, (2) citric acid cycle in the aerobic respiration, and (3) extracellular exosome signaling. These findings may shed light on new directions to understand the mechanism of MDD, potentially facilitating the development of probiotics for better psychiatric disorder treatment. 1 Introduction 1.1 Major Depressive Disorder Major Depressive Disorder (MDD) is a mood disorder that will affect the mood, behavior and other physical parts.
    [Show full text]
  • The Hydroxyquinoline Analog YUM70 Inhibits GRP78 to Induce ER Stress- Mediated Apoptosis in Pancreatic Cancer
    Author Manuscript Published OnlineFirst on February 2, 2021; DOI: 10.1158/0008-5472.CAN-20-1540 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. The hydroxyquinoline analog YUM70 inhibits GRP78 to induce ER stress- mediated apoptosis in pancreatic cancer Soma Samanta1, Suhui Yang1, Bikash Debnath1, Ding Xue1, Yuting Kuang1, 2, Kavya Ramkumar1, 2, Amy S. Lee3, Mats Ljungman4, and Nouri Neamati1* 1Department of Medicinal Chemistry, College of Pharmacy, Rogel Cancer Center, University of Michigan, Ann Arbor, MI, 48109, USA. 2Department of Pharmacology and Pharmaceutical Sciences, School of Pharmacy, University of Southern California, Los Angeles, CA, USA. 3Department of Biochemistry and Molecular Medicine, University of Southern California, Keck School of Medicine, USC Norris Comprehensive Cancer Center, Los Angeles 90089, CA, USA. 4Department of Radiation Oncology, Rogel Cancer Center, Center for RNA Biomedicine, University of Michigan Medical School, and Department of Environmental Health Sciences, School of Public Health, University of Michigan, Ann Arbor, Michigan 48109, USA. *Corresponding Author: Nouri Neamati, Ph.D. Department of Medicinal Chemistry, College of Pharmacy, Rogel Cancer Center, University of Michigan, North Campus Research Complex, 1600 Huron Parkway, Bldg 520, Room 1363, Ann Arbor, MI 48109, Phone: 734-647-2732, Email: [email protected] Running title: YUM70 induces ER stress and apoptosis in pancreatic cancer Keywords: GRP78, ER stress, apoptosis, pancreatic cancer, PROTAC Additional information: Corresponding Author: Nouri Neamati, Ph.D. Department of Medicinal Chemistry, College of Pharmacy, Rogel Cancer Center, University of Michigan, North Campus Research Complex, 1600 Huron PKWY, Bldg 520, Room 1363, Ann Arbor, MI 48109, Phone: 734-647-2732, Email: [email protected] DECLARATION OF INTERESTS: The authors declare no competing interests 1 Downloaded from cancerres.aacrjournals.org on September 25, 2021.
    [Show full text]
  • Revostmm Vol 10-4-2018 Ingles Maquetaciûn 1
    108 ORIGINALS / Rev Osteoporos Metab Miner. 2018;10(4):108-18 Roca-Ayats N1, Falcó-Mascaró M1, García-Giralt N2, Cozar M1, Abril JF3, Quesada-Gómez JM4, Prieto-Alhambra D5,6, Nogués X2, Mellibovsky L2, Díez-Pérez A2, Grinberg D1, Balcells S1 1 Departamento de Genética, Microbiología y Estadística - Facultad de Biología - Universidad de Barcelona - Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER) - Instituto de Salud Carlos III (ISCIII) - Instituto de Biomedicina de la Universidad de Barcelona (IBUB) - Instituto de Investigación Sant Joan de Déu (IRSJD) - Barcelona (España) 2 Unidad de Investigación en Fisiopatología Ósea y Articular (URFOA); Instituto Hospital del Mar de Investigaciones Médicas (IMIM) - Parque de Salud Mar - Centro de Investigación Biomédica en Red de Fragilidad y Envejecimiento Saludable (CIBERFES); Instituto de Salud Carlos III (ISCIII) - Barcelona (España) 3 Departamento de Genética, Microbiología y Estadística; Facultad de Biología; Universidad de Barcelona - Instituto de Biomedicina de la Universidad de Barcelona (IBUB) - Barcelona (España) 4 Unidad de Metabolismo Mineral; Instituto Maimónides de Investigación Biomédica de Córdoba (IMIBIC); Hospital Universitario Reina Sofía - Centro de Investigación Biomédica en Red de Fragilidad y Envejecimiento Saludable (CIBERFES); Instituto de Salud Carlos III (ISCIII) - Córdoba (España) 5 Grupo de Investigación en Enfermedades Prevalentes del Aparato Locomotor (GREMPAL) - Instituto de Investigación en Atención Primaria (IDIAP) Jordi Gol - Centro de Investigación
    [Show full text]