CD300a blockade enhances efferocytosis by infiltrating myeloid cells and ameliorates neuronal deficit after ischemic stroke

Nakahashi-Oda, Chigusa; Fujiyama, Satoshi; Nakazawa, Yuta; Kanemaru, Kazumasa; Wang, Yaqiu; Lyu, Wenxin; Shichita, Takashi; Kitaura, Jiro; Abe, Fumie; Shibuya, Akira

doi:10.1126/sciimmunol.abe7915

Cited by 21 publications

(22 citation statements)

References 68 publications

(85 reference statements)

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“…However, in MAs’ miRNA profile after exosome treatment, we identified three extra negatively related pairs related to the processes of efferocytosis and foam cell differentiation. All three pairs reflected a corresponding change in both mRNA and protein levels (miR-653–5p up vs CD300a down, miR-34b-5p down vs ABR up, miR-344e-3p and 590-3p down vs EP300 up), wherein CD300a-blocking was reported to offset phosphatidylserine binding competition with CD300b and improve CD300b/DAP12-mediated efferocytosis. , Hence, exosomes initiated the efferocytosis repair process through regulating MAs’ mRNA and miRNA expression independently as in Figure .…”

Section: Discussionmentioning

confidence: 92%

“…All three pairs reflected a corresponding change in both mRNA and protein levels (miR-653−5p up vs CD300a down, miR-34b-5p down vs ABR up, miR-344e-3p and 590-3p down vs EP300 up), wherein CD300a-blocking was reported to offset phosphatidylserine binding competition with CD300b and improve CD300b/DAP12-mediated efferocytosis. 46,47 Hence, exosomes initiated the efferocytosis repair process through regulating MAs' mRNA and miRNA expression independently as in Figure 10. In this work, exosome treatment repressed SMCs' proliferation while it enhanced ECs', which is primarily due to exosome's ability to regulate ROS generation.…”

Section: Discussionmentioning

confidence: 98%

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Exosome-Loaded Pro-efferocytic Vascular Stent with Lp-PLA₂-Triggered Release for Preventing In-Stent Restenosis

et al. 2022

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The efferocytosis defect is regarded as a pivotal event of atherosclerosis. The failure to clear apoptotic cells in atherosclerotic plaques under vascular stents causes a failure to resolve the inflammation underneath. However, efferocytosis repair is still confined to nonstenting therapeutics. Here, we identified a pro-efferocytotic agent and accordingly developed a bioresponsive pro-efferocytotic vascular stent aimed for poststenting healing. Exosomes derived from mesenchymal stem cells were found to be able to regulate efferocytosis via SLC2a1, STAT3/RAC1, and CD300a pathways and modulate foam cell formation processes through a CD36-mediated pathway. Pro-efferocytotic exosomes were encapsulated into liposome-based multivesicular chambers and grafted onto vascular stents. The multivesicular vesicles were able to release exosomes under the Lp-PLA2 environment. Compared to bare metal stents, exosome-stents in the presence of Lp-PLA2 enhanced the ratio of apoptotic cell clearance and reduced the neointimal thickness in the mal-efferocytotic rat model. Overall, we identified a pro-efferocytic agentexosomes that are able to regulate target cells via multiple signaling pathways and are good candidates to serve complex pathological environments, and this bioresponsive pro-efferocytotic vascular stent is an attractive approach for prevention of poststenting complications.

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Section: Discussionmentioning

confidence: 92%

Section: Discussionmentioning

confidence: 98%

Exosome-Loaded Pro-efferocytic Vascular Stent with Lp-PLA₂-Triggered Release for Preventing In-Stent Restenosis

et al. 2022

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“…1d ), was identified as a novel negative regulator 23 . Specifically, the binding of an AC with Cd300a and the activation of downstream signaling suppresses efferocytosis by myeloid cells, thus the blockage of Cd300a enhanced efferocytosis 23 . We were also able to validate the results in BMDM using a gRNA targeting Cd300a (Supplementary Fig.…”

Section: Resultsmentioning

confidence: 99%

“…MAGeCK (mageck test) uses Robust Rank Aggregation (RRA) for robust identification of CRISPR-screen hits, and outputs the summary results at both sgRNA and gene level as ranked lists of screen hits. Independent validation of top screen hits by individual gRNAs was performed by lentiviral transduction of gRNA in Rosa-Cas9 knock-in BM cells and differentiation to BMDM followed by efferocytosis assays and quantification 23 .…”

Section: Methodsmentioning

confidence: 99%

A genome-wide CRISPR screen identifies WDFY3 as a regulator of macrophage efferocytosis

Shi

Wang

et al. 2022

Nat Commun

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Phagocytic clearance of dying cells, termed efferocytosis, is essential for maintaining tissue homeostasis, yet our understanding of efferocytosis regulation remains incomplete. Here we perform a FACS-based, genome-wide CRISPR knockout screen in primary mouse macrophages to search for novel regulators of efferocytosis. The results show that Wdfy3 knockout in macrophages specifically impairs uptake, but not binding, of apoptotic cells due to defective actin disassembly. Additionally, WDFY3 interacts with GABARAP, thus facilitating LC3 lipidation and subsequent lysosomal acidification to permit the degradation of apoptotic cell components. Mechanistically, while the C-terminus of WDFY3 is sufficient to rescue the impaired degradation induced by Wdfy3 knockout, full-length WDFY3 is required to reconstitute the uptake of apoptotic cells. Finally, WDFY3 is also required for efficient efferocytosis in vivo in mice and in vitro in primary human macrophages. This work thus expands our knowledge of the mechanisms of macrophage efferocytosis, as well as supports genome-wide CRISPR screen as a platform for interrogating complex functional phenotypes in primary macrophages.

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“…Data were analyzed using MAGeCK (Model-based Analysis of Genome-wide CRISPR-Cas9 Knockout 21 to obtain ranked lists of screen hits. Independent validation of top screen hits by individual gRNAs was performed by lentiviral transduction of gRNA in Rosa-Cas9 knockin BM cells and differentiation to BMDM followed by efferocytosis assays and quantification 23 .…”

Section: Crispr-cas9 Screen For Efferocytosis In Bmdmsmentioning

confidence: 99%

A Genome-wide CRISPR Screen Identifies WDFY3 as a Novel Regulator of Macrophage Efferocytosis

Shi

Wang

et al. 2022

Preprint

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Phagocytic clearance of dying cells, termed efferocytosis, must occur efficiently to maintain homeostasis and prevent disease. Yet, our understanding of this important biological process remains incomplete. To search for novel regulators of efferocytosis, we performed a FACS-based genome-wide CRISPR knockout screen in primary murine macrophages. We identified a novel role for WDFY3 in efferocytosis by macrophages. WDFY3 deficiency in macrophages specifically impaired uptake, not binding, of apoptotic cells due to defective actin depolymerization. We further revealed that WDFY3 directly interacts with GABARAP, thus facilitating LC3 lipidation and subsequent lysosomal acidification to permit the degradation of apoptotic cell components. Although the C-terminus of WDFY3 was sufficient to rescue impaired degradation, full-length WDFY3 is still required for regulating uptake. Finally, WDFY3 is required for efficient efferocytosis in vivo in mice and in primary human macrophages. The work expands our knowledge of the mechanisms of macrophage efferocytosis, and more broadly, provides a general strategy for genome-wide CRISPR screen to interrogate complex functional phenotypes in primary macrophages.

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CD300a blockade enhances efferocytosis by infiltrating myeloid cells and ameliorates neuronal deficit after ischemic stroke

Abstract: CD300a blockade ameliorates stroke pathology through enhancing efferocytosis in the brain by infiltrating myeloid cells.

Cited by 21 publications

References 68 publications

Exosome-Loaded Pro-efferocytic Vascular Stent with Lp-PLA₂-Triggered Release for Preventing In-Stent Restenosis

Exosome-Loaded Pro-efferocytic Vascular Stent with Lp-PLA₂-Triggered Release for Preventing In-Stent Restenosis

A genome-wide CRISPR screen identifies WDFY3 as a regulator of macrophage efferocytosis

A Genome-wide CRISPR Screen Identifies WDFY3 as a Novel Regulator of Macrophage Efferocytosis

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CD300a blockade enhances efferocytosis by infiltrating myeloid cells and ameliorates neuronal deficit after ischemic stroke

Abstract: CD300a blockade ameliorates stroke pathology through enhancing efferocytosis in the brain by infiltrating myeloid cells.

Cited by 21 publications

References 68 publications

Exosome-Loaded Pro-efferocytic Vascular Stent with Lp-PLA2-Triggered Release for Preventing In-Stent Restenosis

Exosome-Loaded Pro-efferocytic Vascular Stent with Lp-PLA2-Triggered Release for Preventing In-Stent Restenosis

A genome-wide CRISPR screen identifies WDFY3 as a regulator of macrophage efferocytosis

A Genome-wide CRISPR Screen Identifies WDFY3 as a Novel Regulator of Macrophage Efferocytosis

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Product

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Exosome-Loaded Pro-efferocytic Vascular Stent with Lp-PLA₂-Triggered Release for Preventing In-Stent Restenosis

Exosome-Loaded Pro-efferocytic Vascular Stent with Lp-PLA₂-Triggered Release for Preventing In-Stent Restenosis