Niemann-Pick disease type C (NPC) is characterized by lysosomal storage of cholesterol and gangliosides, which results from defects in intracellular lipid trafficking. Most studies of NPC1 have focused on its role in intracellular cholesterol movement. Our hypothesis is that NPC1 facilitates the egress of cholesterol from late endosomes, which are where active NPC1 is located. When NPC1 is defective, cholesterol does not exit late endosomes; instead, it is carried along to lysosomal storage bodies, where it accumulates. In this study, we addressed whether cholesterol is transported from endosomes to the plasma membrane before reaching NPC1-containing late endosomes. Our study was conducted in Chinese hamster ovary cell lines that display the classical NPC biochemical phenotype and belong to the NPC1 complementation group. We used three approaches to test whether low density lipoprotein (LDL)-derived cholesterol en route to NPC1-containing organelles passes through the plasma membrane. First, we used cyclodextrins to measure the arrival of LDL cholesterol at the plasma membrane and found that the arrival of LDL cholesterol in a cyclodextrin-accessible pool was significantly delayed in NPC1 cells. Second, the movement of LDL cholesterol to NPC1-containing late endosomes was assessed and found to be normal in Chinese hamster ovary mutant 3-6, which exhibits defective movement of plasma membrane cholesterol to intracellular membranes. Third, we examined the movement of plasma membrane cholesterol to the endoplasmic reticulum and found that this pathway is intact in NPC1 cells, i.e. it does not pass through NPC1-containing late endosomes. Our data suggest that in NPC1 cells LDL cholesterol traffics directly through endosomes to lysosomes, bypassing the plasma membrane, and is trapped there because of dysfunctional NPC1. Niemann-Pick disease type C (NPC)1 is an autosomal recessive lysosomal storage disease that is caused by mutations in either the NPC1 or NPC2 genes and leads to progressive neurodegeneration and premature death (1). Fibroblasts from NPC1 patients exhibit massive storage of unesterified cholesterol and gangliosides in lysosomes, which results from defects in intracellular lipid trafficking (reviewed in Refs. 2 and 3). Most studies of NPC1 have focused on its role in intracellular cholesterol movement. In normal cells, low density lipoproteins (LDL) are internalized and transported through the endocytic pathway where the cholesteryl esters are hydrolyzed. The cholesterol that is released is transported to the plasma membrane and endoplasmic reticulum (4). Our working hypothesis has been that NPC1 facilitates the egress of free cholesterol from late endosomes, which are the location of functional NPC1 (5, 6). When NPC1 is defective, the free cholesterol does not exit the late endosomes; instead, it is carried along the endocytic pathway to lysosomal storage bodies where it accumulates along with mutant NPC1 protein.In this study, we addressed whether cholesterol released by hydrolysis of LDL-derived chol...

We investigated the role of LMNA in adipose tissue by developing a novel mouse model of lipodystrophy. Transgenic mice were generated that express the LMNA mutation that causes familial partial lipodystrophy of the Dunnigan type (FPLD2). The phenotype observed in FPLD-transgenic mice resembles many of the features of human FPLD2, including lack of fat accumulation, insulin resistance, and enlarged, fatty liver. Similar to the human disease, FPLD-transgenic mice appear to develop normally, but after several weeks they are unable to accumulate fat to the same extent as their wild-type littermates. One poorly understood aspect of lipodystrophies is the mechanism of fat loss. To this end, we have examined the effects of the FPLD2 mutation on fat cell function. Contrary to the current literature, which suggests FPLD2 results in a loss of fat, we found that the key mechanism contributing to the lack of fat accumulation involves not a loss, but an apparent inability of the adipose tissue to renew itself. Specifically, preadipocytes are unable to differentiate into mature and fully functional adipocytes. These findings provide insights not only for the treatment of lipodystrophies, but also for the study of adipogenesis, obesity, and insulin resistance.-Wojtanik, K. M., K. Edgemon, S. Viswanadha, B. Lindsey, M. Haluzik, W. Chen, G. Poy, M. Reitman, and C. Londos. The role of LMNA in adipose: a novel mouse model of lipodystrophy based on the Dunnigan-type familial partial lipodystrophy mutation.

What is it? Niemann-Pick C1 (NPC1) is a 1278 amino acid membrane protein residing in the endosomal-lysosomal system. Studies in humans and animal models substantiate NPC1 as a crucial player in intracellular cholesterol trafficking. Can we live without it?Mutations in the NPC1 gene result in Niemann-Pick C disease, an autosomal recessive lipid storage disease. Affected patients display progressive neurodegeneration beginning in early childhood, typically leading to death during the teen years. How did it get its name?The first Niemann-Pick patient was described in 1914 by German pediatrician, Albert Niemann, who noted similarities to Gaucher disease. Ludwig Pick later determined that the disease was distinct from Gaucher, and modestly called the disease Niemann-Pick.