Microbial Pathogenesis in the Era of Spatial Omics

The biology of a cell, whether it is a unicellular organism or part of a multicellular network, is influenced by cell type, temporal changes in cell state, and the cell’s environment. Spatial cues play a critical role in the regulation of microbial pathogenesis strategies. ABSTRACT The biology of a cell, whether it is a unicellular organism or part of a multicellular network, is influenced by cell type, temporal changes in cell state, and the cell’s environment. Spatial cues play a critical role in the regulation of microbial pathogenesis strategies. Information about where the pathogen is—in a tissue or in proximity to a host cell—regulates gene expression and the compartmentalization of gene products in the microbe and the host. Our understanding of host and pathogen identity has bloomed with the accessibility of transcriptomics and proteomics techniques. A missing piece of the puzzle has been our ability to evaluate global transcript and protein expression in the context of the subcellular niche, primary cell, or native tissue environment during infection. This barrier is now lower with the advent of new spatial omics techniques to understand how location regulates cellular functions. This review will discuss how recent advances in spatial proteomics and transcriptomics approaches can address outstanding questions in microbial pathogenesis.

[1]  S. Bullman,et al.  Effect of the intratumoral microbiota on spatial and cellular heterogeneity in cancer , 2022, Nature.

[2]  Jocelyn Y. Kishi,et al.  Light-Seq: light-directed in situ barcoding of biomolecules in fixed cells and tissues for spatially indexed sequencing , 2022, Nature Methods.

[3]  T. Otto,et al.  Single cell and spatial transcriptomic analyses reveal microglia-plasma cell crosstalk in the brain during Trypanosoma brucei infection , 2022, Nature Communications.

[4]  A. Brunner,et al.  Unbiased spatial proteomics with single-cell resolution in tissues. , 2022, Molecular cell.

[5]  B. Budnik,et al.  A Proximity biotinylation assay with a host protein bait reveals multiple factors modulating enterovirus replication , 2022, bioRxiv.

[6]  A. Brunner,et al.  Deep Visual Proteomics defines single-cell identity and heterogeneity , 2022, Nature Biotechnology.

[7]  Valesca Anschau,et al.  A BioID-Derived Proximity Interactome for SARS-CoV-2 Proteins , 2022, Viruses.

[8]  Xianting Ding,et al.  The Intriguing Landscape of Single‐Cell Protein Analysis , 2022, Advanced science.

[9]  Madhav Mantri,et al.  Spatiotemporal transcriptomics reveals pathogenesis of viral myocarditis , 2021, bioRxiv.

[10]  Evan Z. Macosko,et al.  High-resolution Slide-seqV2 spatial transcriptomics enables discovery of disease-specific cell neighborhoods and pathways , 2021, bioRxiv.

[11]  V. Espina,et al.  Laser Capture Proteomics: spatial tissue molecular profiling from the bench to personalized medicine , 2021, Expert review of proteomics.

[12]  C. Blish,et al.  The proximal proteome of 17 SARS-CoV-2 proteins links to disrupted antiviral signaling and host translation , 2021, PLoS pathogens.

[13]  Dexter Pratt,et al.  A BioID-derived proximity interactome for SARS-CoV-2 proteins , 2021, bioRxiv.

[14]  Sarah E. Ewald,et al.  Automated Spatially Targeted Optical Microproteomics Investigates Inflammatory Lesions In Situ , 2021, Journal of proteome research.

[15]  Y. Kozorovitskiy,et al.  Cell-type and subcellular compartment-specific APEX2 proximity labeling reveals activity-dependent nuclear proteome dynamics in the striatum , 2021, Nature Communications.

[16]  L. Sibley,et al.  Toxoplasma gondii secreted effectors co-opt host repressor complexes to inhibit necroptosis. , 2021, Cell host & microbe.

[17]  Vijay Kumar,et al.  Single-Cell Transcriptomics: Current Methods and Challenges in Data Acquisition and Analysis , 2021, Frontiers in Neuroscience.

[18]  Andy S. Moon,et al.  Identification and Molecular Dissection of IMC32, a Conserved Toxoplasma Inner Membrane Complex Protein That Is Essential for Parasite Replication , 2021, mBio.

[19]  S. Carr,et al.  Proximity-Labeling Reveals Novel Host and Parasite Proteins at the Toxoplasma Parasitophorous Vacuole Membrane , 2021, bioRxiv.

[20]  S. P. Singh,et al.  In vivo proximity labeling identifies cardiomyocyte protein networks during zebrafish heart regeneration , 2021, bioRxiv.

[21]  Bryan D. Bryson,et al.  Single Cell and Spatial Transcriptomics Defines the Cellular Architecture of the Antimicrobial Response Network in Human Leprosy Granulomas , 2020, bioRxiv.

[22]  Aditi S Kesari,et al.  A Novel Proximity Biotinylation Assay Based on the Self-Associating Split GFP1–10/11 , 2020, Proteomes.

[23]  P. Hotez,et al.  Host Immunity and Inflammation to Pulmonary Helminth Infections , 2020, Frontiers in Immunology.

[24]  C. Thaiss,et al.  Host-Microbiome Interactions in the Era of Single-Cell Biology , 2020, Frontiers in Cellular and Infection Microbiology.

[25]  Oliver M. Crook,et al.  A Comprehensive Subcellular Atlas of the Toxoplasma Proteome via hyperLOPIT Provides Spatial Context for Protein Functions , 2020, Cell host & microbe.

[26]  C. Romanoski,et al.  Transcriptional Profiling Suggests T Cells Cluster around Neurons Injected with Toxoplasma gondii Proteins , 2020, mSphere.

[27]  L. Sibley,et al.  Toxoplasma gondii secreted effectors co-opt host repressor complexes to inhibit necroptosis , 2020, bioRxiv.

[28]  D. Schüler,et al.  Towards standardized purification of bacterial magnetic nanoparticles for future in vivo applications. , 2020, Acta biomaterialia.

[29]  Michael J. Sweredoski,et al.  Tyramide signal amplification mass spectrometry (TSA-MS) ratio identifies nuclear speckle proteins , 2020, The Journal of cell biology.

[30]  Howard Y. Chang,et al.  RNA-GPS Predicts SARS-CoV-2 RNA Residency to Host Mitochondria and Nucleolus , 2020, Cell Systems.

[31]  K. Roux,et al.  Comparative Application of BioID and TurboID for Protein-Proximity Biotinylation , 2020, Cells.

[32]  D. Gilbert,et al.  SPIN reveals genome-wide landscape of nuclear compartmentalization , 2020, Genome Biology.

[33]  B. Gamain,et al.  An Exported Kinase Family Mediates Species-Specific Erythrocyte Remodelling and Virulence in Human Malaria , 2020, Nature Microbiology.

[34]  A. Lamond,et al.  Quantitative Profiling of the Human Substantia Nigra Proteome from Laser-capture Microdissected FFPE Tissue , 2020, Molecular & Cellular Proteomics.

[35]  Andy S. Moon,et al.  Ancient MAPK ERK7 is regulated by an unusual inhibitory scaffold required for Toxoplasma apical complex biogenesis , 2020, Proceedings of the National Academy of Sciences.

[36]  P. Ivanov,et al.  Spatio-temporal Proteomic Analysis of Stress Granule Disassembly Using APEX Reveals Regulation by SUMOylation and Links to ALS Pathogenesis , 2020, bioRxiv.

[37]  Sarah E. Ewald,et al.  Automated Spatially Targeted Optical Micro Proteomics (autoSTOMP) to Determine Protein Complexity of Subcellular Structures. , 2019, Analytical chemistry.

[38]  A. Belmont,et al.  TSA-seq reveals a largely conserved genome organization relative to nuclear speckles with small position changes tightly correlated with gene expression changes , 2019, Genome research.

[39]  S. Quake,et al.  Cell-Surface Proteomic Profiling in the Fly Brain Uncovers Wiring Regulators , 2019, Cell.

[40]  Patrik L. Ståhl,et al.  High-definition spatial transcriptomics for in situ tissue profiling , 2019, Nature Methods.

[41]  M. Schwartz,et al.  Laser capture microdissection coupled mass spectrometry (LCM-MS) for spatially resolved analysis of formalin-fixed and stained human lung tissues , 2019, bioRxiv.

[42]  D. Humphreys,et al.  A robust fractionation method for protein subcellular localization studies in Escherichia coli. , 2019, BioTechniques.

[43]  Evan Z. Macosko,et al.  Slide-seq: A scalable technology for measuring genome-wide expression at high spatial resolution , 2019, Science.

[44]  R. Satija,et al.  Integrative single-cell analysis , 2019, Nature Reviews Genetics.

[45]  Oliver M. Crook,et al.  Combining LOPIT with differential ultracentrifugation for high-resolution spatial proteomics , 2019, Nature Communications.

[46]  Michael Howell,et al.  Defining host–pathogen interactions employing an artificial intelligence workflow , 2019, eLife.

[47]  Howard Y. Chang,et al.  Atlas of Subcellular RNA Localization Revealed by APEX-Seq , 2018, Cell.

[48]  Nicholas T. Ingolia,et al.  Proximity RNA labeling by APEX-Seq Reveals the Organization of Translation Initiation Complexes and Repressive RNA Granules , 2018, bioRxiv.

[49]  Jian Ma,et al.  Mapping 3D genome organization relative to nuclear compartments using TSA-Seq as a cytological ruler , 2018, The Journal of cell biology.

[50]  N. Perrimon,et al.  Efficient proximity labeling in living cells and organisms with TurboID , 2018, Nature Biotechnology.

[51]  M. L. Previti,et al.  Compartment-Specific Labeling of Bacterial Periplasmic Proteins by Peroxidase-Mediated Biotinylation. , 2018, ACS infectious diseases.

[52]  R. Bataller,et al.  Ductular Reaction Cells Display an Inflammatory Profile and Recruit Neutrophils in Alcoholic Hepatitis , 2018, Hepatology.

[53]  F. Collins,et al.  Biotinylation by antibody recognition - A method for proximity labeling , 2017, Nature Methods.

[54]  S. Teichmann,et al.  A practical guide to single-cell RNA-sequencing for biomedical research and clinical applications , 2017, Genome Medicine.

[55]  Laurent Gatto,et al.  Using hyperLOPIT to perform high-resolution mapping of the spatial proteome , 2017, Nature Protocols.

[56]  Andy S. Moon,et al.  Novel insights into the composition and function of the Toxoplasma IMC sutures , 2017, Cellular microbiology.

[57]  James P. Carson,et al.  Spatially-Resolved Proteomics: Rapid Quantitative Analysis of Laser Capture Microdissected Alveolar Tissue Samples , 2016, Scientific Reports.

[58]  I. Cristea,et al.  A Portrait of the Human Organelle Proteome In Space and Time during Cytomegalovirus Infection. , 2016, Cell systems.

[59]  Patrik L. Ståhl,et al.  Visualization and analysis of gene expression in tissue sections by spatial transcriptomics , 2016, Science.

[60]  Kenneth H. Roux,et al.  An improved smaller biotin ligase for BioID proximity labeling , 2016, Molecular biology of the cell.

[61]  M. Kowshik,et al.  Techniques for the Isolation of Magnetotactic Bacteria , 2016 .

[62]  Qi Liu,et al.  Integrative Omics Analysis Reveals Post-Transcriptionally Enhanced Protective Host Response in Colorectal Cancers with Microsatellite Instability , 2015, Journal of proteome research.

[63]  A. Emili,et al.  Determining composition of micron-scale protein deposits in neurodegenerative disease by spatially targeted optical microproteomics , 2015, eLife.

[64]  Valerie Le Sage,et al.  Proteomic analysis of HIV-1 Gag interacting partners using proximity-dependent biotinylation , 2015, Virology Journal.

[65]  Yanhui Hu,et al.  Proteomic mapping in live Drosophila tissues using an engineered ascorbate peroxidase , 2015, Proceedings of the National Academy of Sciences.

[66]  C. Robinson,et al.  Using proximity biotinylation to detect herpesvirus entry glycoprotein interactions: Limitations for integral membrane glycoproteins. , 2015, Journal of virological methods.

[67]  S. Neil,et al.  Serine Phosphorylation of HIV-1 Vpu and Its Binding to Tetherin Regulates Interaction with Clathrin Adaptors , 2015, PLoS pathogens.

[68]  Weiyong Shen,et al.  Laser capture microdissection: from its principle to applications in research on neurodegeneration , 2015, Neural regeneration research.

[69]  J. Ravel,et al.  SINC, a type III secreted protein of Chlamydia psittaci, targets the inner nuclear membrane of infected cells and uninfected neighbors , 2015, Molecular biology of the cell.

[70]  Evan Z. Macosko,et al.  Highly Parallel Genome-wide Expression Profiling of Individual Cells Using Nanoliter Droplets , 2015, Cell.

[71]  Vaibhav Upadhyay,et al.  Protein recovery from inclusion bodies of Escherichia coli using mild solubilization process , 2015, Microbial Cell Factories.

[72]  Andy S. Moon,et al.  Novel Components of the Toxoplasma Inner Membrane Complex Revealed by BioID , 2015, mBio.

[73]  Kun Zhang,et al.  Fluorescent in situ sequencing (FISSEQ) of RNA for gene expression profiling in intact cells and tissues , 2015, Nature Protocols.

[74]  E. Barklis,et al.  Analysis of HIV-1 Gag Protein Interactions via Biotin Ligase Tagging , 2015, Journal of Virology.

[75]  Mark H. Ellisman,et al.  Directed evolution of APEX2 for electron microscopy and proteomics , 2014, Nature Methods.

[76]  Emily J. Kabeiseman,et al.  The eukaryotic signal sequence, YGRL, targets the chlamydial inclusion , 2014, Front. Cell. Infect. Microbiol..

[77]  S. Carr,et al.  Proteomic mapping of the human mitochondrial intermembrane space in live cells via ratiometric APEX tagging. , 2014, Molecular cell.

[78]  V. Doye,et al.  Probing nuclear pore complex architecture with proximity-dependent biotinylation , 2014, Proceedings of the National Academy of Sciences.

[79]  George M. Church,et al.  Highly Multiplexed Subcellular RNA Sequencing in Situ , 2014, Science.

[80]  D. Foster,et al.  Laser microdissection coupled with RNA-seq analysis of porcine enterocytes infected with an obligate intracellular pathogen (Lawsonia intracellularis) , 2013, BMC Genomics.

[81]  K. Djinović-Carugo,et al.  Novel Bilobe Components in Trypanosoma brucei Identified Using Proximity-Dependent Biotinylation , 2012, Eukaryotic Cell.

[82]  B. Maček,et al.  Analysis of the Plasmodium falciparum proteasome using Blue Native PAGE and label-free quantitative mass spectrometry , 2012, Amino Acids.

[83]  Hyungwon Choi,et al.  SAINT: Probabilistic Scoring of Affinity Purification - Mass Spectrometry Data , 2010, Nature Methods.

[84]  D. Linke,et al.  Efficient subfractionation of gram-negative bacteria for proteomics studies. , 2010, Journal of proteome research.

[85]  Min Jae Lee,et al.  Trimming of Ubiquitin Chains by Proteasome-associated Deubiquitinating Enzymes* , 2010, Molecular & Cellular Proteomics.

[86]  Hyung-Hwan Kim,et al.  Prevalence of Plasmodium vivax VK210 and VK247 subtype in Myanmar , 2010, Malaria Journal.

[87]  M. Shirakawa,et al.  Structural basis for the multiple interactions of the MyD88 TIR domain in TLR4 signaling , 2009, Proceedings of the National Academy of Sciences.

[88]  S. Lemieux,et al.  The phagosomal proteome in interferon-gamma-activated macrophages. , 2009, Immunity.

[89]  P. Nordenfelt,et al.  Isolation of bacteria-containing phagosomes by magnetic selection , 2008, BMC Cell Biology.

[90]  D. Roos,et al.  Organellar dynamics during the cell cycle of Toxoplasma gondii , 2008, Journal of Cell Science.

[91]  C. Bertozzi,et al.  Redirecting lipoic acid ligase for cell surface protein labeling with small-molecule probes , 2007, Nature Biotechnology.

[92]  Conrad Bessant,et al.  Quantitative proteomic approach to study subcellular localization of membrane proteins , 2006, Nature Protocols.

[93]  E. Petricoin,et al.  Laser Capture Microdissection , 1996, Science.

[94]  Xiaohui S. Xie,et al.  A Mammalian Organelle Map by Protein Correlation Profiling , 2006, Cell.

[95]  Rod B. Watson,et al.  Localization of Organelle Proteins by Isotope Tagging (LOPIT)*S , 2004, Molecular & Cellular Proteomics.

[96]  D. Swenson,et al.  Isolation of Mycobacterium tuberculosis mutants defective in the arrest of phagosome maturation. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[97]  M. Muroi,et al.  MD-2 Is Necessary for the Toll-Like Receptor 4 Protein To Undergo Glycosylation Essential for Its Translocation to the Cell Surface , 2003, Clinical Diagnostic Laboratory Immunology.

[98]  W. Souza,et al.  Cell fractionation of parasitic protozoa: a review. , 2003 .

[99]  T. B. Taylor,et al.  High-quality RNA from cells isolated by laser capture microdissection. , 2002, BioTechniques.

[100]  S. Henikoff,et al.  Identification of in vivo DNA targets of chromatin proteins using tethered Dam methyltransferase , 2000, Nature Biotechnology.

[101]  J. Burgess,et al.  New developments in the analysis of gene expression , 2000, Redox report : communications in free radical research.

[102]  P. Cossart,et al.  A comparative study of the actin-based motilities of the pathogenic bacteria Listeria monocytogenes, Shigella flexneri and Rickettsia conorii. , 1999, Journal of cell science.

[103]  Robert F. Bonner,et al.  Laser Capture Microdissection: Molecular Analysis of Tissue , 1997, Science.

[104]  M B Arnao,et al.  Inactivation of peroxidase by hydrogen peroxide and its protection by a reductant agent. , 1990, Biochimica et biophysica acta.

[105]  D. Warhurst,et al.  SEPARATION OF MALARIA-INFECTED ERYTHROCYTES FROM WHOLE BLOOD: USE OF A SELECTIVE HIGH-GRADIENT MAGNETIC SEPARATION TECHNIQUE , 1981, The Lancet.

[106]  R. R. Bensley,et al.  Studies on cell structure by the freezing‐drying method V. The chemical basis of the organization of the cell , 1934 .

[107]  A. Belmont,et al.  Measuring Cytological Proximity of Chromosomal Loci to Defined Nuclear Compartments with TSA-seq. , 2022, Methods in molecular biology.

[108]  L. Sherwood,et al.  Visualizing Filoviral Nucleoproteins Using Nanobodies Fused to the Ascorbate Peroxidase Derivatives APEX2 and dEAPX. , 2022, Methods in molecular biology.

[109]  Sebastian J. Markmiller,et al.  APEX Proximity Labeling of Stress Granule Proteins. , 2022, Methods in molecular biology.

[110]  R. Peek,et al.  Pathobiology of Helicobacter pylori-Induced Gastric Cancer. , 2016, Gastroenterology.

[111]  L. Liotta,et al.  Laser capture microdissection for protein and NanoString RNA analysis. , 2013, Methods in molecular biology.

[112]  M. Howarth,et al.  Imaging proteins in live mammalian cells with biotin ligase and monovalent streptavidin , 2008, Nature Protocols.

[113]  W. de Souza,et al.  Cell fractionation of parasitic protozoa: a review. , 2003, Memorias do Instituto Oswaldo Cruz.