Dataset Information

Phagocyte Transcriptomic Analysis Reveals Focal Adhesion Kinase (FAK) and Heparan Sulfate Proteoglycans (HSPGs) as Major Regulators in Anti-bacterial Defense of Crassostrea hongkongensis.

ABSTRACT: Invertebrates generally lack adaptive immunity and compensate for this with highly efficient innate immune machineries such as phagocytosis by hemocytes to eradicate invading pathogens. However, how extrinsically cued hemocytes marshal internal signals to accomplish phagocytosis is not yet fully understood. To this end, we established a facile magnetic cell sorting method to enrich professional phagocytes from hemocytes of the Hong Kong oyster (Crassostrea hongkongensis), an ecologically and commercially valuable marine invertebrate. Transcriptomic analysis on presorted cells shows that phagocytes maintain a remarkable array of differentially expressed genes that distinguish them from non-phagocytes, including 352 significantly upregulated genes and 479 downregulated genes. Pathway annotations reveal that focal adhesion and extracellular matrix-receptor interactions were the most conspicuously enriched pathways in phagocytes. Phagocytosis rate dramatically declined in the presence of an FAK inhibitor, confirming importance of the focal adhesion pathway in regulating phagocytosis. In addition, we also found that heparan sulfate proteoglycan (HSPG) families were lineage-specifically expanded in C. hongkongensis and abundantly expressed in phagocytes. Efficiency of phagocytosis and hemocytes aggregation was markedly reduced upon blockage of endogenous synthesis of HSPGs, thus implicating these proteins as key surface receptors in pathogen recognition and initiation of phagocytosis.

SUBMITTER: Lin Y

PROVIDER: S-EPMC7103635 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Phagocyte Transcriptomic Analysis Reveals Focal Adhesion Kinase (FAK) and Heparan Sulfate Proteoglycans (HSPGs) as Major Regulators in Anti-bacterial Defense of <i>Crassostrea hongkongensis</i>.

Lin Yue Y Mao Fan F Wong Nai-Kei NK Zhang Xiangyu X Liu Kunna K Huang Minwei M Ma Haitao H Xiang Zhiming Z Li Jun J Xiao Shu S Zhang Yang Y Yu Ziniu Z

Frontiers in immunology 20200320

Invertebrates generally lack adaptive immunity and compensate for this with highly efficient innate immune machineries such as phagocytosis by hemocytes to eradicate invading pathogens. However, how extrinsically cued hemocytes marshal internal signals to accomplish phagocytosis is not yet fully understood. To this end, we established a facile magnetic cell sorting method to enrich professional phagocytes from hemocytes of the Hong Kong oyster (<i>Crassostrea hongkongensis</i>), an ecologically ...[more]

PMID: 32265912

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Project description:BackgroundInterstitial flow directly affects cells that reside in tissues and regulates tissue physiology and pathology by modulating important cellular processes including proliferation, differentiation, and migration. However, the structures that cells utilize to sense interstitial flow in a 3-dimensional (3D) environment have not yet been elucidated. Previously, we have shown that interstitial flow upregulates matrix metalloproteinase (MMP) expression in rat vascular smooth muscle cells (SMCs) and fibroblasts/myofibroblasts via activation of an ERK1/2-c-Jun pathway, which in turn promotes cell migration in collagen. Herein, we focused on uncovering the flow-induced mechanotransduction mechanism in 3D.Methodology/principal findingsCleavage of rat vascular SMC surface glycocalyx heparan sulfate (HS) chains from proteoglycan (PG) core proteins by heparinase or disruption of HS biosynthesis by silencing N-deacetylase/N-sulfotransferase 1 (NDST1) suppressed interstitial flow-induced ERK1/2 activation, interstitial collagenase (MMP-13) expression, and SMC motility in 3D collagen. Inhibition or knockdown of focal adhesion kinase (FAK) also attenuated or blocked flow-induced ERK1/2 activation, MMP-13 expression, and cell motility. Interstitial flow induced FAK phosphorylation at Tyr925, and this activation was blocked when heparan sulfate proteoglycans (HSPGs) were disrupted. These data suggest that HSPGs mediate interstitial flow-induced mechanotransduction through FAK-ERK. In addition, we show that integrins are crucial for mechanotransduction through HSPGs as they mediate cell spreading and maintain cytoskeletal rigidity.Conclusions/significanceWe propose a conceptual mechanotransduction model wherein cell surface glycocalyx HSPGs, in the presence of integrin-mediated cell-matrix adhesions and cytoskeleton organization, sense interstitial flow and activate the FAK-ERK signaling axis, leading to upregulation of MMP expression and cell motility in 3D. This is the first study to describe a flow-induced mechanotransduction mechanism via HSPG-mediated FAK activation in 3D. This study will be of interest in understanding the flow-related mechanobiology in vascular lesion formation, tissue morphogenesis, cancer cell metastasis, and stem cell differentiation in 3D, and also has implications in tissue engineering.

Dataset Information

Phagocyte Transcriptomic Analysis Reveals Focal Adhesion Kinase (FAK) and Heparan Sulfate Proteoglycans (HSPGs) as Major Regulators in Anti-bacterial Defense of Crassostrea hongkongensis.

Publications

Phagocyte Transcriptomic Analysis Reveals Focal Adhesion Kinase (FAK) and Heparan Sulfate Proteoglycans (HSPGs) as Major Regulators in Anti-bacterial Defense of <i>Crassostrea hongkongensis</i>.

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