Adhesion, motility and matrix-degrading gene expression changes in CSF-1-induced mouse macrophage differentiation

Abstract

Migratory macrophages play critical roles in tissue development, homeostasis and disease so it is important to understand how their migration machinery is regulated. Whole transcriptome sequencing revealed that CSF-1-stimulated differentiation of bone marrow-derived precursors into mature macrophages is accompanied by widespread, profound changes in expression of genes regulating adhesion, actin cytoskeletal remodeling and extracellular matrix degradation. Significantly altered expression of almost 40% of adhesion genes, 60 to 86% of Rho family GTPases, their regulators and effectors and over 70% of extracellular proteases occurred. The gene expression changes were mirrored by changes in macrophage adhesion associated with increases in motility and matrix degrading capacity. IL-4 further increased motility and matrix degrading capacity in mature macrophages with additional changes in migration machinery gene expression. Finally, siRNA-induced reductions in the expression of core adhesion proteins, paxillin and leupaxin, decreased macrophage spreading and adhesion numbers with distinct effects on adhesion numbers and their distribution and on matrix degradation. Together, the datasets provide an important resource for our understanding of the regulation of migration in macrophages and to develop therapies targeting disease-enhancing macrophages.

Withholding of M-CSF Supplement Reprograms Macrophages to M2-Like via Endogenous CSF-1 Activation

“…To further validate our findings, macrophages from P3 versus P4 were analyzed in GO pathways. Consistent with the previous report that showed the development of an adherent phenotype of BMDMs upon exposure to increasing doses of M-CSF [40], the GO pathways from our data also manifested the cells from P4 versus P3 fractions in the regulation of cell motility, cell adhesion and cell migration (Figure 6D). Those higher activities indicates faithfully the M2-like phenotypes of macrophages.…”

“…In contrast to the preponderance of extracellular matrix related makers in the upregulated gene set, the top downregulated genes in P4 macrophages encoded proteins with disparate functions, including biological process (CLEC4A1, ERMARD), glycolytic process (ENO1B), metabolic process (DYNLT1B, ATP6V0C-PS2, FUNDC2), cell cycle process (CCNA2, PAGR1A, CDK20, HJURP, DLGAP5), mRNA processing (SRSF7), cellular response (IFI209), response to virus (OAS1A, EVI2), defense response (CCR5, HIST1H2BK) and unknown (BCC003331, CYP4F16). Consistent with previous study [40], we found that the top upregulated Gene Ontology (GO) pathways regulated cell motility, cell adhesion and cell migration mostly (Figure 5D) and the top downregulated GO pathways regulated metabolism, cell cycle and cellular response, and so forth. (Figure 5E).…”

Therapeutic inhibition of the SRC-kinase HCK facilitates T cell tumor infiltration and improves response to immunotherapy

“…While we cannot formally exclude legacy activity of RK20449 against FGR, our previous genetic observations in Hck KO hosts argue that the antitumor effects conferred by RK20449 arise primarily from limiting HCK activity ( 3 ). We also note that FGR is less abundant than HCK in myeloid cells and down-regulated along the monocyte-macrophage differentiation pathway ( 34 ).…”

The Culture Dish Surface Influences the Phenotype and Dissociation Strategy in Distinct Mouse Macrophage Populations

“…Both membrane protein integrity and cell viability during dissociation are critical for successful downstream biological studies ( 3 , 4 ), including flow cytometry and single cell RNA sequencing. Besides, cell adhesion characteristics vary with macrophage maturation/polarization ( 5 , 6 ). Therefore, it is essential to determine the appropriate dissociation methods for different macrophage populations.…”