Characterization and expansion of regulatory natural killer cells for the therapy of chronic graft-versus-host disease
department or school: Medicine, Faculty of
department or school: Pathology and Laboratory Medicine, Department of
data provider: DSpace
degree campus: UBCV
creator: Lauener, Madeline
issued: 2024-07-02T17:01:22Z
issued: 2024
related degree: Doctor of Philosophy - PhD
degree grantor: University of British Columbia
description: Chronic graft-versus-host disease (cGvHD) occurs in ~25% of pediatric and ~60% of adult Hematopoietic Stem Cell Transplant (HSCT) patients and is a major cause of mortality. In large study cohorts (~500 patients), our group has shown a CD56ᵇʳⁱᵍʰᵗPerforin⁻ Natural Killer population, consistent with described regulatory NK cells (NKreg), to strongly associate with protection from cGvHD development. Therefore, I hypothesized that NKreg cells suppress cGvHD through inflammatory cell inhibition, and these cells can be distinguished as a unique cell subset which can be expanded. In my project, I characterized NKreg cells according to transcriptome, proteome, metabolome, phenotype, and function; evaluated the in vitro suppressive effect of NKreg cells on cGvHD immune populations; and optimized the expansion of functional NKreg cells in vitro. Transcriptome analysis revealed that NKreg cells express high RNA levels of Granzyme K, IL-7R, GPR183, RANK, GM-CSFR, CD62L, and LEF1. These NKreg cells can be isolated by sorting for CD56⁺CD16⁻ NK cells, and this population can suppress allogeneic CD4⁺ T cells, but not Treg cells, CD8⁺ T cells, CD56ᵈⁱᵐCD16⁺ cytolytic NK cells, or B cells, through a contact-dependent mechanism, which is partially reliant on the PD-1, LAG-3, and/or TRAIL inhibitory pathways. Additionally, NKreg cells demonstrate a lack of cytotoxicity towards K562, MOLT-4, and Jurkat leukemic cells, and CD4⁺ T cells. Further, I determined two alternative approaches for obtaining clinically relevant numbers of NKreg cells, achieving up to 300-fold expansion, with use of IL-2, TGF-β1, and rapamycin for cells from peripheral blood, or approximately 350-fold expansion when expanding and differentiating NKreg cells from cord blood-hematopoietic stem and progenitor cells. In further characterization of the expanded NKreg cells, I describe their metabolism to be consistent with cytolytic NK cells, though they also secrete regulatory cytokines, including IL-10, TGF-β1, adenosi…
aggregated CHO: https://circle.library.ubc.ca/rest/handle/2429/88549
note: CHARACTERIZATION AND EXPANSION OF REGULATORY NATURAL KILLER CELLS FOR THE THERAPY OF CHRONIC GRAFT-VERSUS-HOST DISEASE by MADELINE LAUENER B.Sc., Simon Fraser University, 2020 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY in THE FACULTY OF GRADUATE AND POSTDOCTORAL STUDIES (Pathology and Laboratory Medicine) THE UNIVERSITY OF BRITISH COLUMBIA (Vancouver) June 2024 © Madeline Lauener, 2024 ii The following individuals certify that they have read, and recommend to the Faculty of Graduate and Postdoctoral Studies for acceptance, the dissertation entitled: Characterization and expansion of regulatory natural killer cells for the therapy of chronic graft-versus-host disease submitted by Madeline Lauener in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Pathology and Laboratory Medicine Examining Committee: Kirk Schultz, MD, Professor, Department of Pediatrics, UBC Supervisor Kevin Hay, MD, Associate Professor, Department of Medicine, University of Calgary Cumming School of Medicine, Department of Medicine Supervisory Committee Member Gregor Reid, PhD, Associate Professor, Department of Pediatrics, UBC University Examiner Alexander Beristain, PhD, Associate Professor, Department of Obstetrics and Gynaecology, UBC University Examiner Denis-Claude Roy, MD, Professor, Department of Medicine, Université de Montréal External Examiner Additional Supervisory Committee Members: Laura Sly, PhD, Professor, Department of Pediatrics, UBC Supervisory Committee Member Ramon Klein Geltink, PhD, Assistant Professor, Department of Pathology and Laboratory Medicine, UBC Supervisory Committee Member iii Abstract Chronic graft-versus-host disease (cGvHD) occurs in ~25% of pediatric and ~60% of adult Hematopoietic Stem Cell Transplant (HSCT) patients and is a major cause of mortality. In large study cohorts (~500 patients), our group has shown a CD56brightPerforin- Natural Killer population, consistent wi…
has type: Thesis/Dissertation
date issued: 2024-11
is shown at: 10.14288/1.0444063
language: eng
degree discipline: Pathology and Laboratory Medicine
provider: Vancouver : University of British Columbia Library
publisher: University of British Columbia
rights: Attribution-NonCommercial-NoDerivatives 4.0 International
rights URI: http://creativecommons.org/licenses/by-nc-nd/4.0/
scholar level: Graduate
contributor: Schultz, Kirk R.
type: Text
identifier URI: http://hdl.handle.net/2429/88549