Single-cell transcriptomic profiling of lung endothelial cells identifies dynamic inflammatory and regenerative subpopulations

Zhang, Lianghui; Gao, Shangce; White, Zachary; Dai, Yang; Malik, Asrar B.; Rehman, Jalees

doi:10.1172/jci.insight.158079

Cited by 34 publications

(28 citation statements)

References 52 publications

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“…In this study, we uncovered a novel pro-inflammatory role of Pink1-mediated mitophagy in endothelial cells. Inflammatory activation induced mitophagy in endothelial cells, both in vitro and in an in vivo endotoxin model of lung injury that is characterized by excessive endothelial inflammation and influx of neutrophils (Bachmaier, Toya et al 2007, Kolaczkowska and Kubes 2013, Zhang, Gao et al 2022). The observed endothelial mitophagy was mediated by Pink1 activation.…”

Section: Discussionmentioning

confidence: 99%

Pink1-mediated mitophagy in the endothelium releases proteins encoded by mitochondrial DNA and activates neutrophil responses

Gajwani

Wang

Srivastava

et al. 2022

Preprint

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Given their ancient evolutionary origins, eukaryotic mitochondria possess multiple vestiges of their prokaryotic ancestors. One such factor is the N-terminal formylation of proteins encoded by mitochondrial DNA. N-formylated proteins are also released by bacteria and trigger activation of immune cells such as neutrophils. Growing evidence indicate that circulating levels of mitochondrial formyl proteins are elevated in the serum of patients with excessive inflammatory responses and trigger neutrophil activation like their bacterial counterparts. However, the cellular source of these proteins, and the mechanism by which they are released into the circulation is not known. In this study, we have identified vascular endothelial cells as a source of mitophagy induced release of formyl proteins in response to inflammatory mediators in vitro. Mechanistically, endothelial mitophagy required activation of the Pink1 pathway. Using liposomal delivery of sgRNA targeting Pink1 in mice expressing endothelial-specific Cas9, we developed a mouse model in which Pink1 is specifically depleted in the endothelium. Deletion of endothelial Pink1 was remarkably protective in endotoxin-induced lung inflammation, resulting in reduced neutrophil infiltration and significantly reduced death in mice. We thus propose that endothelial cells upregulate pro-inflammatory mitophagy in response to inflammation, leading to release of mitochondrial formyl peptides and detrimental neutrophil recruitment into the lung.

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Section: Discussionmentioning

confidence: 99%

Pink1-mediated mitophagy in the endothelium releases proteins encoded by mitochondrial DNA and activates neutrophil responses

Gajwani

Wang

Srivastava

et al. 2022

Preprint

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“…The mice were monitored every other day to measure tumor growth with electronic caliper and body weight for 3 weeks and tumor size was presented with tumor volume which was calculated with equation of width 2 x height x 0.523 as previously described 78 . For lineage tracing of muscle endothelial cells during distal tumor growth, we used inducible endothelial specific tdTomato expressing mice (tdTomato flfl:Cdh5-CreERT2, EC tdTomato ) 49 . To induce the Cre recombinase in tdTomato fl/fl: Cdh5-CreERT2 mice, 8 weeks EC tdTomato mice were intraperitoneally administrated with tamoxifen (100 mg/kg, body weight, Sigma-Aldrich T5648) once a day for consecutive three days.…”

Section: Methodsmentioning

confidence: 99%

“…Thus, we examined whether cachexic muscles have specific EC subpopulations that drive muscle loss using scRNA-seq analysis. We took advantage of inducible EC specific tdTomato lineage tracing mice (Cdh5-Cre ERT2 -tdTomato, EC tdTomato ) 49 which would maintain their irreversible genetic endothelial lineage label with tdTomato. The skeletal muscles from the hindlimbs of tumor free EC tdTomato mice (WT) and melanoma-cachexia EC tdTomato mice were used to isolate the tdTomato positive (+) cells (ECs) and tdTomato negative (-) cells (non-ECs) by flow cytometric sorting and the isolated cells were subjected to scRNA-seq using the 10X Genomics 3' platform.…”

Section: Single Cell Transcriptomics Identifies Hypoxia Catabolism An...mentioning

confidence: 99%

Impaired Barrier Integrity of the Skeletal Muscle Vascular Endothelium Drives Progression of Cancer Cachexia

Kim

Sanborn

Wang

et al. 2022

Preprint

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Cancer patients experience cachexia, which is characterized by extensive skeletal muscle wasting that worsens the quality of life and increases mortality. Currently, there are no approved treatments that can effectively counteract cancer cachexia. Vascular endothelial cells (ECs) are essential for maintaining tissue perfusion, nutrient supply, and preventing inappropriate transmigration of immune cells into the tissue. However, little is known about the role of the muscle vasculature in cancer cachexia. We hypothesized that endothelial dysfunction in the skeletal muscle mediates cancer cachexia. Using transgenic pancreatic ductal adenocarcinoma (PDAC) mice and a tissue clearing and high-resolution 3D-tissue imaging approach, we found that the loss of skeletal muscle vascular density precedes the loss of muscle mass. Importantly, we show that cancer cachexia patients exhibit significantly decreased muscle vascular density and severe muscle atrophy when compared to non-cancer patients. Unbiased single cell transcriptomic analyses of the muscle endothelium unveiled a unique EC population present in cachexia muscles. Increased circulating Activin-A suppresses the expression of the transcriptional co-activator PGC1alpha in the muscle endothelium, thus disrupting junctional integrity in the vasculature and increasing vascular leakage. Conversely, restoration of endothelial-specific PGC1alpha prevented the decreased vascular density and muscle loss observed in tumor-bearing mice. Our study suggests that EC-PGC1alpha is essential for maintaining the integrity of the skeletal muscle vascular barrier and that restoring muscle endothelial function could be a valuable therapeutic approach to prevent or reverse cancer cachexia.

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“…The mice were monitored every other day to measure tumor growth with electronic caliper and body weight for 3 weeks and tumor size was presented with tumor volume which was calculated with equation of width 2 x height x 0.523 as previously described 78 . For lineage tracing of muscle endothelial cells during distal tumor growth, we used inducible endothelial speci c tdTomato expressing mice (tdTomato :Cdh5-CreERT2, EC tdTomato ) 49 . To induce the Cre recombinase in tdTomato / : Cdh5-CreERT2 mice, 8 weeks EC tdTomato mice were intraperitoneally administrated with tamoxifen (100 mg/kg, body weight, Sigma-Aldrich T5648) once a day for consecutive three days.…”

Section: Methodsmentioning

confidence: 99%

Impaired Barrier Integrity of the Skeletal Muscle Vascular Endothelium Drives Progression of Cancer Cachexia

Rehman

Kim

Sanborn

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

Cancer patients experience cachexia, which is characterized by extensive skeletal muscle wasting that worsens the quality of life and increases mortality. Currently, there are no approved treatments that can effectively counteract cancer cachexia. Vascular endothelial cells (ECs) are essential for maintaining tissue perfusion, nutrient supply, and preventing inappropriate transmigration of immune cells into the tissue. However, little is known about the role of the muscle vasculature in cancer cachexia. We hypothesized that endothelial dysfunction in the skeletal muscle mediates cancer cachexia. Using transgenic pancreatic ductal adenocarcinoma (PDAC) mice and a tissue clearing and high-resolution 3D-tissue imaging approach, we found that the loss of skeletal muscle vascular density precedes the loss of muscle mass. Importantly, we show that cancer cachexia patients exhibit significantly decreased muscle vascular density and severe muscle atrophy when compared to non-cancer patients. Unbiased single cell transcriptomic analyses of the muscle endothelium unveiled a unique EC population present in cachexia muscles. Increased circulating Activin-A suppresses the expression of the transcriptional co-activator PGC1α in the muscle endothelium, thus disrupting junctional integrity in the vasculature and increasing vascular leakage. Conversely, restoration of endothelial-specific PGC1α prevented the decreased vascular density and muscle loss observed in tumor-bearing mice. Our study suggests that endothelial PGC1α is essential for maintaining the integrity of the skeletal muscle vascular barrier and that restoring muscle endothelial function could be a valuable therapeutic approach to prevent or reverse cancer cachexia.

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Single-cell transcriptomic profiling of lung endothelial cells identifies dynamic inflammatory and regenerative subpopulations

Cited by 34 publications

References 52 publications

Pink1-mediated mitophagy in the endothelium releases proteins encoded by mitochondrial DNA and activates neutrophil responses

Pink1-mediated mitophagy in the endothelium releases proteins encoded by mitochondrial DNA and activates neutrophil responses

Impaired Barrier Integrity of the Skeletal Muscle Vascular Endothelium Drives Progression of Cancer Cachexia

Impaired Barrier Integrity of the Skeletal Muscle Vascular Endothelium Drives Progression of Cancer Cachexia

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