title: Rapid and label-free microfluidic neutrophil purification and phenotyping in diabetes mellitus.
abstract: Advanced management of dysmetabolic syndromes such as diabetes will benefit from a timely mechanistic insight enabling personalized medicine approaches. Herein, we present a rapid microfluidic neutrophil sorting and functional phenotyping strategy for type 2 diabetes mellitus (T2DM) patients using small blood volumes (fingerprick ~100 μL). The developed inertial microfluidics technology enables single-step neutrophil isolation (>90% purity) without immuno-labeling and sorted neutrophils are used to characterize their rolling behavior on E-selectin, a critical step in leukocyte recruitment during inflammation. The integrated microfluidics testing methodology facilitates high throughput single-cell quantification of neutrophil rolling to detect subtle differences in speed distribution. Higher rolling speed was observed in T2DM patients (P < 0.01) which strongly correlated with neutrophil activation, rolling ligand P-selectin glycoprotein ligand 1 (PSGL-1) expression, as well as established cardiovascular risk factors (cholesterol, high-sensitive C-reactive protein (CRP) and HbA1c). Rolling phenotype can be modulated by common disease risk modifiers (metformin and pravastatin). Receiver operating characteristics (ROC) and principal component analysis (PCA) revealed neutrophil rolling as an important functional phenotype in T2DM diagnostics. These results suggest a new point-of-care testing methodology, and neutrophil rolling speed as a functional biomarker for rapid profiling of dysmetabolic subjects in clinical and patient-oriented settings.
authors.affiliation: Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore.
authors.forename: Han Wei
authors.initials: HW
authors.lastname: Hou
authors.affiliation: Mechanical and Aerospace Engineering, Nanyang Technological University, Singapore.
authors.forename: Chayakorn
authors.initials: C
authors.lastname: Petchakup
authors.affiliation: Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore.
authors.forename: Hui Min
authors.initials: HM
authors.lastname: Tay
authors.affiliation: Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore.
authors.forename: Zhi Yang
authors.initials: ZY
authors.lastname: Tam
authors.affiliation: Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore.|Endocrine and Diabetes, Tan Tock Seng Hospital, Singapore.
authors.forename: Rinkoo
authors.initials: R
authors.lastname: Dalan
authors.affiliation: Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore.|Endocrine and Diabetes, Tan Tock Seng Hospital, Singapore.
authors.forename: Daniel Ek Kwang
authors.initials: DE
authors.lastname: Chew
authors.affiliation: Mechanical and Aerospace Engineering, Nanyang Technological University, Singapore.
authors.forename: King Ho Holden
authors.initials: KH
authors.lastname: Li
authors.affiliation: Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore.|Endocrine and Diabetes, Tan Tock Seng Hospital, Singapore.|Imperial College London, UK.
authors.forename: Bernhard O
authors.initials: BO
authors.lastname: Boehm
mesh_terms: D000328:Adult; D002469:Cell Separation / Q000379:methods*; D003924:Diabetes Mellitus, Type 2 / Q000175:diagnosis / Q000276:immunology*; D006801:Humans; D036904:Leukocyte Rolling; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D008875:Middle Aged; D009504:Neutrophils / Q000166:cytology*; D010641:Phenotype; D025341:Principal Component Analysis; D012372:ROC Curve; D059010:Single-Cell Analysis; D055815:Young Adult