Biology subjects

Kislev, N.

Publications and source records attributed to Kislev, N..

4 recordsLinked to original sources

bioRxiv2026

SVEP1 couples adipose niche remodeling to macrophage activation and impaired metabolism in obesity

The extracellular matrix (ECM) of adipose tissue regulates metabolic homeostasis, yet how ECM composition drives adipose dysfunction in obesity remains only partially defined. Multi-omics profiling of visceral adipose tissue identified the multidomain ECM protein SVEP1 as a candidate regulator of adipose remodeling; SVEP1 was elevated in obese human adipose tissue and correlated with insulin resistance and systemic inflammation, and interaction analysis predicted associations with ECM proteins, integrins, and cytoskeletal regulators. On a chow diet, Svep1 heterozygous (Svep1+/-) mice gained less weight with lower fasting glucose, and their adipocytes showed enhanced lipolysis and AMPK activation. On a high-fat diet, Svep1+/- mice exhibited attenuated adiposity, improved glucose tolerance and insulin sensitivity, and fewer adipose macrophages with a shift away from a pro-inflammatory profile, alongside downregulation of inflammatory, lipid-metabolic, and ECM-remodeling pathways. Decellularized Svep1+/- matrix altered macrophage adhesion and phenotype relative to Svep1+/+ matrix, identifying SVEP1 as a matrix-borne mediator coupling adipose remodeling to immune activation and metabolic impairment. Together, these findings position the adipose ECM as an active driver of immunometabolic dysfunction and nominate SVEP1 and its receptor network as candidate targets in obesity.

cell biology↗

bioRxiv2023

Advance Glycation End-products induction accelerate amyloid deposits in adipocyte`s lipid droplets

Adipose tissue dysfunction is central to insulin resistance, and the emergence of type 2 diabetes (T2D) is associated with elevated levels of carbonyl metabolites from glucose metabolism. In this study, using methylglyoxal (MGO) and glycolaldehyde (GAD) carbonyl metabolites, induced protein glycation leading to misfolding and {beta}-sheet formation and generation of advanced glycation end products (AGEs). The formed AGEs compromise adipocytes activity. Microscopic and spectroscopic assays were used to examine the impact of MGO and GAD on lipid droplet - associated proteins. The results provide information about how glycation leads to the appearance of amyloidogenic proteins formation that hinders metabolism and autophagy in adipocytes. We measured the beneficial effects of metformin, an anti-diabetic drug, on misfolded protein as assessed by thioflavin (ThT) spectroscopy and improved autophagy. In vitro findings were complemented by in vivo analysis of white adipose tissue (WAT), where lipid droplet-associated {beta}-amyloid deposits were predominantly linked to adipose triglyceride lipase (ATGL), a lipid droplet protein. Bioinformatics, imaging, and biochemical methods affirm ATGLs role in {beta}-sheet secondary structure creation. Our results highlighted the pronounced presence of amyloidogenic proteins in adipocytes treated with carbonyl compounds, potentially reshaping our understanding of adipocyte pathology in the context of T2D. This in-depth exploration offers novel perspectives on related pathophysiology and underscores the potential of adipocytes as pivotal therapeutic targets, bridging T2D, amyloidosis, protein glycation, and adipocyte malfunction. Significance StatementThe generation of advanced glycation end products (AGEs) has a strong connection to diabetes severity . Adipose tissue is known to play a key role in the metabolic impairment and obesity associated with diabetes. We used the carbonyl compounds methylglyoxal (MGO) and glycolaldehyde (GAD) to create AGEs in adipocytes. The results of this study indicate that glycation not only affects cell metabolism and impairs adipocyte lipolysis, but also alters autophagy and increases protein amyloid deposits related to the membrane of lipid droplets. We identify the ATGL as a protein prone to {beta} sheet alteration. consequently, ATGL emerges as a pivotal actor in lipid droplet metabolism and a prospective therapeutic target for T2D complications.

cell biology↗

bioRxiv2021

MYH10 governs adipocyte function and adipogenesis through its interaction with GLUT4

Adipocyte differentiation is dependent on cytoskeletal remodeling processes that determine and maintain cellular shape and function. In turn, cytoskeletal proteins contribute to the filament-based network responsible for controlling adipocytes shape and promoting the intracellular trafficking of key cellular components. Currently, our understanding of these mechanisms remains incomplete. In this study, we identified the non-muscle myosin 10 (MYH10) as an important regulator of adipogenesis and adipocyte function through its interaction with the insulin dependent, Glucose transporter 4 (GLUT4). MYH10 depletion in preadipocytes resulted in impaired adipogenesis, with knockdown cells exhibiting disrupted morphology and reduced molecular adipogenic signals. MYH10 was shown to be in complex with GLUT4 in adipocytes, an interaction regulated by insulin induction. The missing adipogenic capacity of MYH10-KD cells was restored when they uptook GLUT4 vesicles up from neighbor wild-type cells in a co-culture system. Our results provide the first demonstration that MYH10 interacts with GLUT4 in cells and adipose tissue through the insulin pathway. The signaling cascade is regulated by the protein kinase C {zeta} (PKC{zeta}), which interacts with MYH10 to modify the localization and interaction of both GLUT4 and MYH10 in adipocytes as PKC{zeta} inhibition resulted in reduced GLUT4 and MYH10 translocation and interactions. Overall, our study establishes MYH10 as an essential regulator of GLUT4 translocation, affecting both adipogenesis and adipocyte function, highlighting its importance in future cytoskeleton-based studies in adipocytes.

cell biology↗

bioRxiv2021

Imaging analysis to quantitate the Interplay of membrane and cytoplasm protein dynamics

Plasma membrane proteins are extremely important in cell signaling and cellular functions. Protein expression and localization alter in response to various signals in a way that is dependent on cell type and niche. Compartmental quantification of the expression of particular proteins is a very useful means of understanding their role in cellular processes. Immunofluorescence staining is frequently used to investigate the distribution of proteins of interest. Here, we present an imaging method for quantifying the membrane to cytoplasm ratio (MCR) of proteins analyzed at single-cell resolution. This technique provides a robust quantification of membrane proteins and contributes new insights into membrane expression dynamics. We have developed a protocol that uses immunostaining to assess protein expression according to the fluorescent cellular distribution and to compute the MCR. The method was applied to measure the MCR of glucose transporter 4 (GLUT4) in response to insulin in 3T3-L1 cells, an in-vitro model for adipocyte function and adipogenesis. The results revealed informative changes in the subcellular localization of GLUT4 following insulin induction. MCR analysis is a powerful imaging tool that can be generally applied to membrane proteins to provide a rapid and efficient quantitative analysis of protein distribution and sub-cellular processes in cells.

cell biology↗