Multiparameter Single Cell Profiling of Airway Inflammatory Cells

Abstract

Airway diseases affect over 7% of the U.S. population and millions of patients worldwide. Asthmatic patients have wide variation in clinical severity with different clinical and physiologic manifestations of disease that may be driven by distinct biologic mechanisms. Further, the immunologic underpinnings of this complex trait disease are heterogeneous and treatment success depends on defining subgroups of asthmatics. Due to the limited availability and number of cells from the lung, the active site, in-depth investigation has been challenging. Recent advances in technology support transcriptional analysis of cells from induced sputum. Flow cytometry studies have described cells present in the sputum but a detailed analysis of these subsets is lacking. Mass cytometry or CyTOF (Cytometry by Time-Of-Flight) offers tremendous opportunities for multiparameter single cell analysis. Experiments can now allow detection of up to ~40 markers to facilitate unprecedented multidimensional cellular analyses. Here we demonstrate the use of CyTOF on primary airway samples obtained from well-characterized patients with asthma and cystic fibrosis. Using this technology, we quantify cellular frequency and functional status of defined cell subsets. Our studies provide a blueprint to define the heterogeneity among subjects and underscore the power of this single cell method to characterize airway immune status.

Single Cell Transcriptional Archetypes of Airway Inflammation in Cystic Fibrosis

“…Nine subjects with a confirmed CF diagnosis from the Yale Adult CF Program provided sputum samples. We also recruited five HC to undergo sputum induction according to previous protocols (22). Study subjects were closely age-matched, with a higher inclusion of female subjects in the CF group (67% CF, n=6, 40% HC, n=2).…”

The end of omics? High dimensional single cell analysis in precision medicine

“…One of the most prominent application of HDcyto technology is the investigation of human pathologies, as it allows a deep understanding of inter‐cellular perturbations and the identification of disease‐specific signatures that may serve as diagnostic biomarkers or, eventually, therapeutic targets. HDcyto has already proved to be a powerful platform for the investigation of idiopathic syndromes and pathologies with incomplete understanding of disease‐driving mechanisms .…”

“…Distinct monocyte/macrophage and granulocyte cell populations distinguished asthmatic patients from cystic fibrosis patients and healthy controls. The method proved to be especially useful for dissecting a heterogeneous condition like asthma into groups with different immunological characteristics which, might reflect differences in disease pathophysiologies and could help to make individualized therapeutic decisions in the future .…”

The road ahead: Implementing mass cytometry in clinical studies, one cell at a time

“…We highlight six studies that illustrate recent progress. The manuscripts by Yao et al , Corneau et al , and Strauss‐Albee et al emphasize the utility of mass cytometry for identifying cell subsets that capture patient‐specific disease attributes in the fields of pulmonology, neonatology, and virology. The studies by Abraham et al and Vendrame et al apply visualization and statistical tools in novel combinations to interpret high‐dimensional data, while the manuscript by Leelatian et al describes a standardized protocol to derive isolated cancer cells from solid tissue for analysis by mass cytometry.…”

“…Yao et al demonstrate the utility of multiparameter profiling of airway inflammatory cells in patients with cystic fibrosis (CF) to better understand the complex interactions between an individual's immune state and disease severity. Using a novel assay for the mass cytometry analysis of airway inflammatory cells, the authors identified significant differences in the frequency of immune cell subsets in sputum collected from patients with CF, patients with asthma, and healthy controls.…”