Work Package 3.
Spatial and temporal regulation of endo-lysosomal processes in lipid metabolism.
Low-density lipoprotein (LDL) cholesterol is taken up by the cell through the LDL receptor (LDLR), a process highly dependent on the endo-lysosomal system. Endocytosed cholesterol is further transported to other cellular compartments. Changes in intracellular cholesterol concentrations modulate endo-lysosomal processes at a spatial and temporal level to maintain cholesterol homeostasis. Defects in these processes are correlated with impaired cholesterol uptake and intracellular transport, which can lead to hypercholesterolemia, cholesterol storage diseases and NAFLD. This work package will investigate the protein networks controlling endosomal cholesterol transport and lipid metabolism and their spatio-temporal regulation in living cells.
PhD projects in this Work Package:
PhD student 4 – Uncovering the spatio-temporal regulation of endo-lysosomal lipid trafficking in control of metabolism
University Medical Centre Utrecht, NL Judith Klumperman, j.klumperman[at]umcutrecht.nlThe PhD student will establish the role of Vps3, Vps8 and Vps33B in cholesterol and LDLR/LRP1 transport and whether the SNX17 retriever CCC complex exit from early endosomes sorts cargo into Vps3/8/33B dependent transport vesicles. The student will be involved in characterizing the compartments and mechanisms involved in segregation of cholesterol/Rab8/CD63 positive carriers from LAMP-positive lysosomes.
PhD student 5 – Coordination of endo-lysosomal cargo transport by NPC1 and post-NPC1 regulators of lipid metabolism
University of Helsinki, FI Elina Ikonen, elina.ikonen[at]helsinki.fiThe PhD student will investigate how trafficking of NPC1 is regulated by endosomal machineries and how this will impact endo-lysosomal lipid delivery to post-NPC1 destinations, including the plasma membrane, ER and lipid droplets. The student will take advantage of advanced live cell imaging techniques to visualize lipids and proteins and dissect how proteins coordinate cellular lipid deposition between endo-lysosomes and lipid droplets, with relevance to lysosomal storage and fatty liver diseases.
PhD student 6 – Genome-wide functional genetic approaches to identify novel regulators of the endo-lysosomal network in control of lipid metabolism
Amsterdam Medical Centre, NL Noam Zelcer, n.zelcer[at]amsterdamumc.nlThe PhD student will develop and use mammalian haploid- and CRISPR-Cas9-based genome-wide genetic screens to interrogate regulation of endogenously tagged proteins involved in intracellular trafficking of cholesterol within the endo-lysosomal network and in delivery of lipoprotein-derived cholesterol to the ER/lipid droplets. Identified regulators will be studied using cellular and mouse models, and the metabolic consequence of their hepatic-specific deficiency on lipid and energy metabolism in atherosclerosis and metabolic syndrome will be investigated.