Generation and Functional Characterization of Integrin αM-Deficient Hoxb8 Hematopoietic Progenitors as a Model for Primary Leukocytes
Abstract
Leukocyte recruitment is a critical component of the immune response, but the over-accumulation of these cells drives the pathology of numerous inflammatory diseases. The αMβ2 (CD11b/CD18) integrin subunit αM serves as a key regulator in this process, mediating leukocyte adhesion, migration, and signal transduction pathways for precise cellular response. Despite its importance as a therapeutic target, studying integrin function presents significant methodological challenges. Currently, integrin function is mostly studied through primary leukocytes or immortalized myeloid cell lines. While primary leukocytes are physiologically accurate, they are short-lived, difficult to genetically manipulate, and limited by ethical constraints. Conversely, myeloid cell lines provide scalability but are susceptible to mutations altering physiological behavior. These limitations can be overcome using Hox transcription factors, which immortalize progenitors while maintaining characteristics identical to primary cells. However, studies generating a functional αM-deficient Hoxb8 cell line are nonexistent. This project aims to establish and characterize an αM-deficient Hoxb8 progenitor cell line, isolated from αM-knockout mice, differentiate the cells, and test their physiological similarity to primary αM-knockout leukocytes. We hypothesize that αM-deficient Hoxb8 hematopoietic progenitors can be differentiated into neutrophils and macrophages, and that resulting cells will exhibit cell-surface receptor expression and adhesion mechanisms similar to primary αM-knockout leukocytes. Wild-type and αM-knockout progenitors will be isolated and conditionally immortalized using the ER-Hoxb8 method. Differentiation will be induced by removing estrogen and introducing leukocyte-specific cytokines. Accurate differentiation will be confirmed via flow cytometry analysis of cell-surface markers. A functional adhesion assay using integrin-specific ligands will test adhesion between wild-type and αM-knockout Hoxb8-derived cells. Based on previous studies with primary and myeloid cell lines, we expect the Hoxb8-derived cells to differentiate into αM-deficient leukocytes and exhibit reduced adhesion to αM-dependent ligands compared to wild-type counterparts. Ultimately, this project will establish a defined, scalable model for studying integrin-dependent leukocyte migration in inflammatory diseases.
Start Time
15-4-2026 9:00 AM
End Time
15-4-2026 10:00 AM
Room Number
272
Presentation Type
Oral Presentation
Presentation Subtype
Research-in-Progress
Presentation Category
Health
Faculty Mentor
Yakubenko Valentin
Generation and Functional Characterization of Integrin αM-Deficient Hoxb8 Hematopoietic Progenitors as a Model for Primary Leukocytes
272
Leukocyte recruitment is a critical component of the immune response, but the over-accumulation of these cells drives the pathology of numerous inflammatory diseases. The αMβ2 (CD11b/CD18) integrin subunit αM serves as a key regulator in this process, mediating leukocyte adhesion, migration, and signal transduction pathways for precise cellular response. Despite its importance as a therapeutic target, studying integrin function presents significant methodological challenges. Currently, integrin function is mostly studied through primary leukocytes or immortalized myeloid cell lines. While primary leukocytes are physiologically accurate, they are short-lived, difficult to genetically manipulate, and limited by ethical constraints. Conversely, myeloid cell lines provide scalability but are susceptible to mutations altering physiological behavior. These limitations can be overcome using Hox transcription factors, which immortalize progenitors while maintaining characteristics identical to primary cells. However, studies generating a functional αM-deficient Hoxb8 cell line are nonexistent. This project aims to establish and characterize an αM-deficient Hoxb8 progenitor cell line, isolated from αM-knockout mice, differentiate the cells, and test their physiological similarity to primary αM-knockout leukocytes. We hypothesize that αM-deficient Hoxb8 hematopoietic progenitors can be differentiated into neutrophils and macrophages, and that resulting cells will exhibit cell-surface receptor expression and adhesion mechanisms similar to primary αM-knockout leukocytes. Wild-type and αM-knockout progenitors will be isolated and conditionally immortalized using the ER-Hoxb8 method. Differentiation will be induced by removing estrogen and introducing leukocyte-specific cytokines. Accurate differentiation will be confirmed via flow cytometry analysis of cell-surface markers. A functional adhesion assay using integrin-specific ligands will test adhesion between wild-type and αM-knockout Hoxb8-derived cells. Based on previous studies with primary and myeloid cell lines, we expect the Hoxb8-derived cells to differentiate into αM-deficient leukocytes and exhibit reduced adhesion to αM-dependent ligands compared to wild-type counterparts. Ultimately, this project will establish a defined, scalable model for studying integrin-dependent leukocyte migration in inflammatory diseases.
