Harnessing lipid signaling pathways to target specialized pro-angiogenic neutrophil subsets for regenerative immunotherapy

Turner, Thomas C.; Sok, Mary Caitlin P.; Hymel, Lauren A.; Pittman, FredE.; York, William Y.; Mac, Quoc D.; Vyshnya, Sofiya; Lim, Hong Seo; Kwong, Gabriel A.; Qiu, Peng; Botchwey, Edward A.

doi:10.1126/sciadv.aba7702

Cited by 15 publications

(12 citation statements)

References 35 publications

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“…Before UMAP dimensional reduction, each flow cytometry sample was manually pregated to select single cells. The gated FCS (flow cytometry standard) files were imported into Python 3.7 using fcsparser Pandas 2.5 according to ( 59 ). Each channel, except for FSC and SSC, was normalized by applying arcsinh transformation with a cofactor of 150 to transform fluorescence data into a fold-channel scale.…”

Section: Methodsmentioning

confidence: 99%

Analyzing immune response to engineered hydrogels by hierarchical clustering of inflammatory cell subsets

et al. 2022

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Understanding the immune response to hydrogel implantation is critical for the design of immunomodulatory biomaterials. To study the progression of inflammation around poly(ethylene glycol) hydrogels presenting Arg-Gly-Asp (RGD) peptides and vascular endothelial growth factor, we used temporal analysis of high-dimensional flow cytometry data paired with intravital imaging, immunohistochemistry, and multiplexed proteomic profiling. RGD-presenting hydrogels created a reparative microenvironment promoting CD206 + cellular infiltration and revascularization in wounded dorsal skin tissue. Unbiased clustering algorithms (SPADE) revealed significant phenotypic transition shifts as a function of the cell-adhesion hydrogel properties. SPADE identified an intermediate macrophage subset functionally regulating in vivo cytokine secretion that was preferentially recruited for RGD-presenting hydrogels, whereas dendritic cell subsets were preferentially recruited to RDG-presenting hydrogels. Last, RGD-presenting hydrogels controlled macrophage functional cytokine secretion to direct polarization and vascularization. Our studies show that unbiased clustering of single-cell data provides unbiased insights into the underlying immune response to engineered materials.

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Section: Methodsmentioning

confidence: 99%

Analyzing immune response to engineered hydrogels by hierarchical clustering of inflammatory cell subsets

et al. 2022

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“…UMAPs generated as previously reported (27, 45, 46). Briefly, UMAP was used to embed high-dimensional flow cytometry data into a space of two dimensions, cells are visualized in a scatter plot where similarity is demonstrated via proximity to other points.…”

Section: Methodsmentioning

confidence: 99%

“…SPADE trees generated as previously reported (27, 45, 46). Briefly, SPADE was performed through MATLAB and the source code is available at http://pengqiu.gatech.edu/software/SPADE/.…”

Section: Methodsmentioning

confidence: 99%

Section: Spanning-tree Progression Analysis Of Density-normalized Events (Spade)mentioning

confidence: 99%

“…A population of cells captured at the same time via high-dimensional immune cell profiling includes many distinct intermediate differentiation states of cells; however, classical analytical techniques only evaluate their average properties and thus mask trends occurring across individual cells and subpopulations (25)(26)(27). In this study, we approached this challenge by employing a unique dimensionality reduction and clustering analytical strategy that combines both uniform manifold approximation and projection (UMAP) and spanning-tree progression analysis of density-normalized events (SPADE); this analytical approach captures the functional heterogeneity of the early immune cell response and dynamics after an injury event and was applied to the scenario of critical VML injury (Figure S1).…”

Section: Introductionmentioning

confidence: 99%

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Identifying dysregulated immune cell subsets following critical volumetric muscle loss with pseudo-time trajectories

Hymel

Turner

et al. 2021

Preprint

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Volumetric muscle loss (VML) results in permanent functional deficits and remains a substantial regenerative medicine challenge. A coordinated immune response is crucial for timely myofiber regeneration, however the immune response following VML has yet to be fully characterized. Here, we leveraged dimensionality reduction and pseudo-time analysis techniques to elucidate the cellular players underlying a functional or pathological outcome as a result of subcritical or critical VML in the murine quadriceps, respectively. We found that critical VML presented with a sustained presence of M2-like and CD206hiLy6Chi "hybrid" macrophages whereas subcritical defects resolved these populations. These macrophage subsets may contribute to fibrogenesis in critical VML, especially in the presence of TGF-β. Furthermore, several T cell populations were significantly elevated in critical VML compared to subcritical injuries. Specifically, there was a significant increase of CD127+ T cells at days 3 and 7, and upregulated CD127 expression may indicate aberrant IL-7 signaling in critical VML. These results demonstrate a dysregulated immune response in critical VML that is unable to resolve the chronic inflammatory state and transition to a pro-regenerative microenvironment. These data provide important insights into potential therapeutic strategies which could reduce the immune cell burden and pro-fibrotic signaling characteristic of VML.

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Effect of stretching on inflammation in a subcutaneous carrageenan mouse model analyzed at single‐cell resolution

Berrueta,

Muñoz‐Vergara,

Martin

et al. 2023

Journal Cellular Physiology

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Understanding the factors that influence the biological response to inflammation is crucial, due to its involvement in physiological and pathological processes, including tissue repair/healing, cancer, infections, and autoimmune diseases. We have previously demonstrated that in vivo stretching can reduce inflammation and increase local pro‐resolving lipid mediators in rats, suggesting a direct mechanical effect on inflammation resolution. Here we aimed to explore further the effects of stretching at the cellular/molecular level in a mouse subcutaneous carrageenan‐inflammation model. Stretching for 10 min twice a day reduced inflammation, increased the production of pro‐resolving mediator pathway intermediate 17‐HDHA at 48 h postcarrageenan injection, and decreased both pro‐resolving and pro‐inflammatory mediators (e.g., PGE2 and PGD2) at 96 h. Single‐cell RNA sequencing analysis of inflammatory lesions at 96 h showed that stretching increased the expression of both pro‐inflammatory (Nos2) and pro‐resolution (Arg1) genes in M1 and M2 macrophages at 96 h. An intercellular communication analysis predicted specific ligand–receptor interactions orchestrated by neutrophils and M2a macrophages, suggesting a continuous neutrophil presence recruiting immune cells such as activated macrophages to contain the antigen while promoting resolution and preserving tissue homeostasis.

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Harnessing lipid signaling pathways to target specialized pro-angiogenic neutrophil subsets for regenerative immunotherapy

Cited by 15 publications

References 35 publications

Analyzing immune response to engineered hydrogels by hierarchical clustering of inflammatory cell subsets

Analyzing immune response to engineered hydrogels by hierarchical clustering of inflammatory cell subsets

Identifying dysregulated immune cell subsets following critical volumetric muscle loss with pseudo-time trajectories

Effect of stretching on inflammation in a subcutaneous carrageenan mouse model analyzed at single‐cell resolution

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