Biology subjects

von Eckardstein, A.

Publications and source records attributed to von Eckardstein, A..

7 recordsLinked to original sources

bioRxiv2026

The protein and lipid compositions of brain apolipoprotein E particles are cell type-specific

Genetic variation in APOE, which encodes a lipid transporter apolipoprotein E (apoE), is the strongest risk factor for late-onset Alzheimer's disease (AD). Although central nervous system (CNS) apoE is produced by various cell types, including astrocytes, microglia, and pericytes, it is unclear whether apoE lipoproteins are a homogeneous population or if their molecular composition varies depending on their cellular origin. Emerging evidence suggests that CNS apoE lipoproteins contribute to AD-related processes, including amyloid deposition, neuroinflammation, synaptic maintenance, and blood-brain barrier function in a cell type-specific manner. We hypothesized that the compositions of CNS apoE lipoproteins depend on their cellular origin and investigated the lipid and protein profiles of apoE particles secreted by induced pluripotent stem cell (iPSC)-derived brain cells and derived from human cerebrospinal fluid (CSF) using liquid chromatography-mass spectrometry (LC-MS/MS). Our results demonstrate that astrocytes, pericytes, and microglia secrete apoE lipoproteins with distinct, cell type-specific lipid and protein profiles. Astrocyte-secreted particles were enriched in sphingomyelins and proteins linked to lipid metabolism, whereas pericyte-secreted particles contained ether-linked phospholipids and proteins associated with antioxidant activity. The lipidation of microglial apoE was markedly lower, but these particles had higher levels of transferrin, apoC-II, and cathepsin D. Notably, one-third of identified features were cell type-specific, highlighting the substantial molecular specialization across cell types. Most lipids and proteins identified in cell-secreted apoE were also detected in CSF-derived particles. Our findings advance the understanding of apoE biology by shifting the focus from apoE as a single, homogeneous entity to a diverse population of complex particles, the composition of which might be linked to functions in health and neurodegenerative diseases.

neuroscience↗

bioRxiv2026

The COPI coatomer influences LDL receptor activity, hepatic lipid storage, and apoB secretion

BackgroundDecreased hepatic removal of low density lipoproteins (LDL) and increased apolipoprotein B (apoB) production cause hypercholesterolemia, a major causal risk factor of atherosclerotic cardiovascular disease (ASCVD). By a genome-wide siRNA screen, we previously identified subunits of the Coat protein I (COPI) complex to limit LDL uptake into Huh-7 hepatocarcinoma cells. MethodsThese findings were validated by targeted in vitro experiments as well as genetic association studies in humans and three mouse models with mutated or disrupted COPI genes. ResultsSilencing of COPA, COPB1, COPB2, ARCN1, COPG1, and COPZ1 in Huh-7 cells resulted in decreased uptake of LDL and aberrant glycosylation and altered cell surface abundance of the LDL receptor (LDLR) as well as increased apoB secretion and cellular lipid storage. Single nucleotide polymorphisms of ARCN1 were associated with lower ARCN1 expression and higher levels of LDL-cholesterol (LDL-C). Rare variants of COPA and COPG1 were enriched among patients with LDL-C > 5 mmol/L. Patients and mice carrying other rare immunopathogenic missense variants of COPA and COPG1 did not present with elevated plasma levels of LDL-C, while hepatic knockdown of murine Copg1 increased the concentrations of non-HDL-cholesterol in plasma and triglycerides in the liver. ConclusionsThe COPI coatomer regulates LDLR activity and apoB secretion as well as lipid content of liver cells. Loss of function of some variants of COPI genes are associated with higher LDL-C levels.

cell biology↗

bioRxiv2025

Angiopoietin like protein 3 regulates low-density lipoprotein transport through aortic endothelial cells via endothelial lipase.

Angiopoietin-like protein 3 (ANGPTL3) inhibits endothelial lipase (EL, encoded by LIPG) and lipoprotein lipase (LPL) and has emerged as a promising target for lipid-lowering therapies. While ANGPTL3 inhibition lowers circulating triglycerides and low-density lipoprotein cholesterol (LDL-C), its impact on atherosclerotic cardiovascular disease (ASCVD) remains uncertain. This study investigates the roles of ANGPTL3 and EL in the trans-endothelial transport of LDL and high-density lipoprotein (HDL), an early step in atherogenesis. Using primary human aortic endothelial cells (HAEC), we demonstrate that EL promotes binding, uptake, and transcytosis of LDL and HDL, with its catalytic activity being essential for transport but not for surface association. ANGPTL3 selectively reduced the transport of LDL but not HDL through HAEC. Mechanistically, EL and scavenger receptor BI (SR-BI) act sequentially to mediate LDL uptake, but independently of each other towards HDL uptake. Ex vivo, ANGPTL3 reduced LDL accumulation within bovine aorta. Interestingly, ANGPTL4 had no measurable effect on lipoprotein transport, while ANGPTL8 modestly inhibited both LDL and HDL association with endothelial cells. Our findings provide evidence that ANGPTL3 inhibits EL-mediated transendothelial LDL transport. Opposing effects of ANGPTL3 inhibition on LDL-C concentration and LDL entry into the arterial wall may explain why recent population studies did not reveal any genetically causal association of ANGPTL3 and LIPG with ASCVD risk.

molecular biology↗

bioRxiv2025

Dyslipidemic SPTLC3 Integrates Bile Acid-FXR Signaling with Sphingolipid Remodeling in MASLD

Molecular mechanisms driving metabolic disease pathogenesis remain poorly understood. Genetic and functional studies implicate SPTLC3 with dyslipidemia. SPTLC3 synthesizes atypical long-chain bases (LCB) precursors for sphingolipid production. We demonstrate significant SPTLC3 expression in human liver. ORMDL1-3 regulate SPTLC3 post-translationally in hepatic and non-hepatic cells. Independently, farnesoid X receptor (FXR) represses hepatic SPTLC3 transcription via a negative promoter element. In mice, high-fat diet (HFD) induced whereas bile acids normalized hepatic SPTLC3 transcription. SPTLC3 derived LCBs in plasma originate from the liver and are elevated in HFD-fed mice and in patients with metabolic dysfunction-associated steatotic liver disease (MASLD). Cross-species comparison revealed marked differences in LCB composition between mice and humans. Notably, omega-3-methylsphingosine (meC18SO) was significantly associated with MASLD in humans but undetectable in mice. In Huh7 cells, meC18SO enhanced complex II and IV activity, oxygen consumption, and mitochondrial ROS content. FXR-SPTLC3 axis and presented findings have potential implications for future translational research.

biochemistry↗

bioRxiv2023

Mapping the dynamic cell surface interactome of high-density lipoprotein reveals Aminopeptidase N as modulator of its endothelial uptake

Heterogeneous high-density lipoprotein (HDL) particles, which can contain hundreds of proteins, affect human health and disease through dynamic molecular interactions with cell surface proteins. How HDL mediates its long-range signaling functions and interactions with various cell types is largely unknown. Due to the complexity of HDL, we hypothesize that multiple receptors engage with HDL particles resulting in condition-dependent receptor-HDL interaction clusters at the cell surface. Here we used the mass spectrometry-based and light-controlled proximity labeling strategy LUX-MS in a discovery-driven manner to decode HDL-receptor interactions. Surfaceome nanoscale organization analysis of hepatocytes and endothelial cells using LUX-MS revealed that the previously known HDL-binding protein scavenger receptor SCRB1 is embedded in a cell surface protein community, which we term HDL synapse. Modulating the endothelial HDL synapse, composed of 60 proteins, by silencing individual members showed that the HDL synapse can be assembled in the absence of SCRB1 and that the members are interlinked. The aminopeptidase AMPN (also known as CD13) was identified as an HDL synapse member that directly influences HDL uptake into the primary human aortic endothelial cells (HAECs). Our data indicate that preformed cell surface residing protein complexes modulate HDL function and suggest new theragnostic opportunities.

systems biology↗

bioRxiv2022

Decoding functional high-density lipoprotein particle surfaceome interactions

High-density lipoprotein (HDL) is a mixture of complex particles mediating reverse cholesterol transport in the human body and several cytoprotective activities. Despite its relevance for human health, many aspects of HDL-mediated lipid trafficking and cellular signaling remain elusive at the molecular level. During HDLs journey throughout the body, its function is mediated through interactions with cell surface receptors on different cell types. Using four different cellular model systems HDL is interacting with, we comparatively analyzed their surfaceomes to define the HDL receptome. Surfaceome analysis of EA.hy926, HEPG2, foam cells, and human aortic endothelial cells (HAEC) revealed the main currently known HDL-receptor scavenger receptor B1 (SCRB1), as well as 154 shared cell surface receptors representing potential HDL interaction receptor candidates. Since vascular endothelial growth factor A (VEGF-A) was recently found as a regulatory factor of transendothelial transport of HDL, we next analyzed the VEGF modulated surfaceome of HAEC using auto-CSC technology. VEGF-A treatment led to a remodeling of the surfaceome of HAEC cells including the previously reported higher surfaceome abundance of SCRB1 upon treatment. 165 additional receptors were found on HAEC upon VEGF-A treatment representing SCRB1 co-regulated receptors potentially involved in HDL function. Using HATRIC-based ligand receptor capturing (HATRIC-LRC) technology on human endothelial cells, we specifically aimed for the identification of other bona fide (co-)receptors of HDL beyond SCRB1. HATRIC-LRC enabled, next to SCRB1, the identification of the receptor tyrosine-protein kinase Mer (MERTK), which we show is directly contributing to endothelial HDL binding and uptake. Subsequent proximity ligation assays (PLA) demonstrate furthermore the spatial vicinity of MERTK and SCRB1 on the endothelial cell surface. The data shown provide direct evidence for a complex and dynamic HDL receptome and that receptor nanoscale organization may influence functional HDL binding and uptake. List of human gene and protein names discussed in the paperABCA1: Phospholipid-transporting ATPase ABCA1 (ABCA1) ABCA7: Phospholipid-transporting ATPase ABCA7 (ABCA7) ABCG1: ATB-binding cassette G1 (ABCG1) ACVR2A: Activin receptor type-2A (AVR2A) APOA1: Apolipoprotein A1 (APOA1) APOB: Apolipoprotein B (APOB) APOC3: Apolipoprotein C-III (APOC3) APOH: Beta-2-glycoprotein 1 (APOH) APOM: Apolipoprotein M (APOM) ATP5MF: Ecto-F1-ATPase (ATPK) CD4: T-cell surface glycoprotein CD4 (CD4) CD14: Monocyte differentiation antigen CD14 (CD14) CD36: platelet glycoprotein 4 (CD36) EGFR: Epidermal growth factor receptor (EGFR) ITGAM: Integrin alpha-M (ITAM) ITGAV: Integrin alpha-V (ITAV) KDR: Vascular endothelial growth factor receptor 2 (VGFR2) KIT: Mast/stem cell growth factor receptor Kit (KIT) MEGF8: Multiple epidermal growth factor-like domains protein 8 (MEGF8) MEGF10: Multiple epidermal growth factor-like domains protein 10 (MEGF10) MERTK: tyrosine-protein kinase Mer (MERTK) MRC2: C-type mannose receptor 2 (MRC2) MSR1: Macrophage scavenger receptor types I and II (MSRE) PCDHAC1: Protocadherin alpha-C1 (PCDC1) PLTP: Phospholipid transfer protein (PLTP) S1PR1: sphingosine 1-phosphate receptor 1 (S1PR1) S1PR2: sphingosine 1-phosphate receptor 2 (S1PR2) S1PR3: sphingosine 1-phosphate receptor 3 (S1PR3) SCARA3: Scavenger receptor class A member 3 (SCAR3) SCARB1: Scavenger receptor B1 (SCRB1) SLC8A3: Sodium/calcium exchanger 2 (NAC2) TF: Transferrin (TRFE) TFRC: Transferrin receptor protein 1 (TFR1) VEGFA: vascular endothelial growth factor A (VEGF-A) Significance statementA molecular understanding of the high-density lipoprotein (HDL) receptome would provide the rational basis for modulating reverse cholesterol transport. Here we used cellular model systems mimicking tissues involved in HDL-trafficking and reverse cholesterol transport to define the HDL receptome. The receptor tyrosine-protein kinase Mer (MERTK) was identified as a novel mediator of HDL binding and uptake in human aortic endothelial cells. Endothelial MERTK resides within the surfaceome proximal to SCRB1 representing a potential co-receptor. The molecular nanoscale organization of the cell-specific surfaceome may influence cellular HDL interactions and functionality.

molecular biology↗

bioRxiv2020

Sphingosine-1-phosphate receptors 1 and 3 regulate the expression of SR-B1 in human aortic endothelial cells

Several vasoprotective functions of high-density lipoproteins (HDL) on the endothelium have been shown to depend on the presence of sphingosine-1-phosphate (S1P) receptors (S1PRs) as well as scavenger receptor class B type 1 (SR-B1). Interference with the presence of S1P or the activity of S1PR1 or S1PR3 mimics many effects seen by the interference with SR-B1. This raises the question on interactions between S1P receptors and SR-B1. We investigated the influence of S1PRs on SR-B1 expression in human aortic endothelial cells. Silencing or pharmacological inhibition of S1PR1 or S1PR3 down-regulated SCARB1 mRNA expression as well as SR-B1 protein abundance. RNA interference with S1PR1 or S1PR3 also decreased cellular association of 125I-HDL with HAECs. Further mechanistic studies showed that knockdown of S1PR1 or S1PR3 reduced SR-B1 protein by inducing its degradation through deceasing Akt activity. Moreover, silencing of S1PR1 or S1PR3 suppressed SCARB1 mRNA expression by decreasing cellular cAMP levels. In conclusion, we provide evidence for an as yet unappreciated interaction, namely the regulation of SR-B1 abundance by S1PRs on both transcriptional and post-translational levels, suggesting that interactions of S1PRs and SR-B1 regulate signaling functions of HDL as well as uptake of lipoproteins in endothelial cells.

cell biology↗