Multiparametric platform for profiling lipid trafficking in human leukocytes

Systematic insight into cellular dysfunction can improve understanding of disease etiology, risk assessment, and patient stratification. We present a multiparametric high-content imaging platform enabling quantification of low-density lipoprotein (LDL) uptake and lipid storage in cytoplasmic droplets of primary leukocyte subpopulations. We validate this platform with samples from 65 individuals with variable blood LDL-cholesterol (LDL-c) levels, including familial hypercholesterolemia (FH) and non-FH subjects. We integrate lipid storage data into another readout parameter, lipid mobilization, measuring the efficiency with which cells deplete lipid reservoirs. Lipid mobilization correlates positively with LDL uptake and negatively with hypercholesterolemia and age, improving differentiation of individuals with normal and elevated LDL-c. Moreover, combination of cell-based readouts with a polygenic risk score for LDL-c explains hypercholesterolemia better than the genetic risk score alone. This platform provides functional insights into cellular lipid trafficking and has broad possible applications in dissecting the cellular basis of metabolic disorders. [Display omitted] •High-content imaging platform for quantification of low-density lipoprotein uptake•Quantifies lipid storage and mobilization in cytoplasmic droplets of primary leukocytes•Approach provides personalized insights into cellular basis of hypercholesterolemia•Combining functional and polygenic scores improves hypercholesterolemia risk assessment We have limited information on how cellular lipid uptake and processing differ between individuals and influence the development of metabolic diseases, such as hypercholesterolemia. Available assays are labor intensive, require skilled personnel, and are difficult to scale to higher throughput, making it challenging to obtain systematic, functional cell-based data from individuals. To overcome this problem, we established a scalable automated analysis pipeline enabling reliable quantification of multiple cellular readouts, including lipid uptake, storage, and mobilization, from different white blood cell populations. This approach provides personalized insights into the cellular basis of hypercholesterolemia and obesity. Insights into cellular dysfunction underlying hypercholesterolemia are lacking. Pfisterer et al. establish an automated analysis platform enabling quantification of multiple cellular readouts, including lipid uptake, storage, and mobili