{"id":{"repo_id":"buffalo","oai_identifier":"oai:ubir.buffalo.edu:10477/83855"},"canonical_url":"https://search.dev.ndltd.org/etd/buffalo/oai:ubir.buffalo.edu:10477/83855","repository":{"repo_id":"buffalo","name":"Buffalo","base_url":"https://ubir.buffalo.edu/oai/request"},"display":{"title":"Molecular Regulators of E-Selectin Mediated Leukocyte Trafficking at Site of Inflammation","abstract":"Ph.D.","abstract_html":"Ph.D.","abstract_has_math":false,"creators":["Zhu, Yuqi"],"institution":"State University of New York at Buffalo","degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["Neelamegham, Sriram","Chemical and Biological Engineering"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2022,"date_issued":"2022-06-17T19:54:49Z","date_published":"2022-06-17T19:54:49Z","updated_at":"2026-07-27T19:05:28Z","subjects":["bioengineering"],"languages":["eng"],"rights":["Users of works found in University at Buffalo Institutional Repository (UBIR) are responsible for identifying and contacting the copyright owner for permission to reuse. University at Buffalo Libraries do not manage rights for copyright-protected works and cannot assist with permissions.","Copyright retained by author."],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/10477/83855","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Neelamegham, Sriram","Chemical and Biological Engineering"]},{"key":"dc:creator","label":"Author","values":["Zhu, Yuqi"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2022-06-17T19:54:49Z","2020"]},{"key":"dc:publisher","label":"Institution","values":["State University of New York at Buffalo"]},{"key":"dc:type","label":"Dc Type","values":["Text","Dissertation"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["bioengineering"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["eng"]},{"key":"dc:rights","label":"Dc Rights","values":["Users of works found in University at Buffalo Institutional Repository (UBIR) are responsible for identifying and contacting the copyright owner for permission to reuse. University at Buffalo Libraries do not manage rights for copyright-protected works and cannot assist with permissions.","Copyright retained by author."]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/10477/83855"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Ph.D.","At site of inflammation, cell adhesion molecules expressed on the surface of endothelial cells including the selectins (E-, P- and L-selectin), the CD18/β2-integrins (LFA-1 and Mac-1) and members of the immunoglobulin/IgG superfamily (ICAMS, JAMs etc.) recruit leukocyte from the flowing blood stream. The transmigration of the recruited leukocytes to the inflamed tissue is mediated by a four step mechanism, including initial leukocyte recruitment/tethering, cell rolling transitioning from rapid to slow rolling, activation primarily via chemokine receptors, and finally firm adhesion & transmigration. These are the fundamental biological steps of immune response towards invading pathogens. Since similar process also mediates cancer metastasis and hematopoietic stem cell homing to bone marrow, it is critical to have a good understanding of the molecular mechanisms regulating this process. Most importantly, the current dissertation focuses on the initial step of this leukocyte adhesion cascade, which involves selectin-ligand interactions. The ligands for selectins are glycoproteins or glycosphingolipids expressed on the surface of the leukocytes. These glyco-conjugates are post-translationally synthesized in Golgi. Various selectin ligands shared similar terminal sialylofucosylated structures with Sialyl LewisX (sLeX) representing a prototypic ligand. However, the core/branch glyco structures and protein substrate can vary among the different selectin-ligands. To study selectin-ligand interaction, there are three major aspects: 1. Identification of the peptide substrate for the ligand; 2. Determination of the carbohydrate structure that attaches to the peptide; 3. Discovering the glycosyltransferases involved in the biosynthesis of relevant glycan determinants. These aspects are covered in this dissertation. While the human L-/P-selectin ligands are already well identified, the ligand for E-selectin still remains unclear. In this thesis, we studied putative human E-selectin ligands, partially using CRISPR-Cas9 based genome editing technology. In this regard, CRISPR-Cas9 is a novel gene manipulation tool to provide high gene editing efficiency, high specificity, and low off-target rate without compromise cell viability. With CRISPR-Cas9, we knocked-out putative E-selectin ligand to determine if the perturbation results in ablated interaction with E-selectin. Also, combined with next-generation sequencing and bioinformatics, CRISPR-Cas9 based whole genome library makes unbiased whole genome screening easy to access. This powerful strategy was applied in all the chapters in this dissertation.","**To request an accessible version of the file(s) associated with this item, contact library@buffalo.edu. Please include the item's persistent URL [http://hdl.handle.net/. . .] in your request.**"]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Molecular Regulators of E-Selectin Mediated Leukocyte Trafficking at Site of Inflammation"]}]}],"canonical_facts":{"dc:contributor":["Neelamegham, Sriram","Chemical and Biological Engineering"],"dc:creator":["Zhu, Yuqi"],"dc:date":["2022-06-17T19:54:49Z","2020"],"dc:description":["Ph.D.","At site of inflammation, cell adhesion molecules expressed on the surface of endothelial cells including the selectins (E-, P- and L-selectin), the CD18/β2-integrins (LFA-1 and Mac-1) and members of the immunoglobulin/IgG superfamily (ICAMS, JAMs etc.) recruit leukocyte from the flowing blood stream. The transmigration of the recruited leukocytes to the inflamed tissue is mediated by a four step mechanism, including initial leukocyte recruitment/tethering, cell rolling transitioning from rapid to slow rolling, activation primarily via chemokine receptors, and finally firm adhesion & transmigration. These are the fundamental biological steps of immune response towards invading pathogens. Since similar process also mediates cancer metastasis and hematopoietic stem cell homing to bone marrow, it is critical to have a good understanding of the molecular mechanisms regulating this process. Most importantly, the current dissertation focuses on the initial step of this leukocyte adhesion cascade, which involves selectin-ligand interactions. The ligands for selectins are glycoproteins or glycosphingolipids expressed on the surface of the leukocytes. These glyco-conjugates are post-translationally synthesized in Golgi. Various selectin ligands shared similar terminal sialylofucosylated structures with Sialyl LewisX (sLeX) representing a prototypic ligand. However, the core/branch glyco structures and protein substrate can vary among the different selectin-ligands. To study selectin-ligand interaction, there are three major aspects: 1. Identification of the peptide substrate for the ligand; 2. Determination of the carbohydrate structure that attaches to the peptide; 3. Discovering the glycosyltransferases involved in the biosynthesis of relevant glycan determinants. These aspects are covered in this dissertation. While the human L-/P-selectin ligands are already well identified, the ligand for E-selectin still remains unclear. In this thesis, we studied putative human E-selectin ligands, partially using CRISPR-Cas9 based genome editing technology. In this regard, CRISPR-Cas9 is a novel gene manipulation tool to provide high gene editing efficiency, high specificity, and low off-target rate without compromise cell viability. With CRISPR-Cas9, we knocked-out putative E-selectin ligand to determine if the perturbation results in ablated interaction with E-selectin. Also, combined with next-generation sequencing and bioinformatics, CRISPR-Cas9 based whole genome library makes unbiased whole genome screening easy to access. This powerful strategy was applied in all the chapters in this dissertation.","**To request an accessible version of the file(s) associated with this item, contact library@buffalo.edu. Please include the item's persistent URL [http://hdl.handle.net/. . .] in your request.**"],"dc:format":["application/pdf"],"dc:identifier":["http://hdl.handle.net/10477/83855"],"dc:language":["eng"],"dc:publisher":["State University of New York at Buffalo"],"dc:rights":["Users of works found in University at Buffalo Institutional Repository (UBIR) are responsible for identifying and contacting the copyright owner for permission to reuse. University at Buffalo Libraries do not manage rights for copyright-protected works and cannot assist with permissions.","Copyright retained by author."],"dc:subject":["bioengineering"],"dc:title":["Molecular Regulators of E-Selectin Mediated Leukocyte Trafficking at Site of Inflammation"],"dc:type":["Text","Dissertation"]},"updated_at":"2026-07-27T19:05:28Z"}