Biology subjects

Stemmer, P.

Publications and source records attributed to Stemmer, P..

4 recordsLinked to original sources

bioRxiv2026

Insulin preservatives trigger neutrophil extracellular trap formation through SYNE1-mediated nuclear disassembly

Neutrophil extracellular traps (NETs) are critical effector molecules in sterile inflammation, yet the molecular mechanisms by which xenobiotic chemical exposures trigger NETosis remain poorly defined. Here, using phenolic preservatives present in all FDA-approved insulin formulations as a discovery platform, we show that these compounds induce NETosis in primary human neutrophils (34.3 {+/-} 5.0% vs. 2.8 {+/-} 0.9% for preservative-free insulin; p < 0.001) via a mechanism distinct from canonical PKC- and calcium-dependent pathways. Data-independent acquisition mass spectrometry (n = 6 donors) reveals that preservatives prompt coordinated dephosphorylation of SYNE1 (nesprin-1) at Ser8724 and Ser8727 (log2FC = -4.41 and -4.01, respectively; both q-value < 0.0001), disrupting LINC complex-mediated nuclear-cytoskeletal anchoring, through a phosphatase-dependent pathway distinct from canonical PKC- and calcium-dependent NETosis. In a porcine subcutaneous catheter model, preservative-containing formulations drive progressive NET accumulation, neutrophil infiltration, and early fibrotic changes over 7 days, whereas removing preservatives reduces the histological inflammation score by 40% (P < 0.001). These findings establish phenolic preservatives as non-pathogen triggers of NETosis, identify disruption of the SYNE1-LINC complex as the underlying mechanism, and demonstrate that preservative-free formulations lessen device-related inflammation, offering a translatable strategy for safer implantable drug delivery systems.

immunology↗

bioRxiv2024

Integrative multiomics analysis of metabolic dysregulation induced by occupational benzene exposure in mice

Background: Type 2 Diabetes Mellitus (T2DM) is a significant public health burden. Emerging evidence links volatile organic compounds (VOCs), such as benzene to endocrine disruption and metabolic dysfunction. However, the effects of chronic environmentally relevant VOC exposures on metabolic health are still emerging. Objective: Building on our previous findings that benzene exposure at smoking levels (50 ppm) induces metabolic impairments in male mice, we investigated the effects of occupationally relevant, below OSHA approved, benzene exposure on metabolic health. Methods: Adult male C57BL/6 mice were exposed to 0.9ppm benzene 8 hours a day for 9 weeks. We assessed measures of metabolic homeostasis and conducted RNA and proteome sequencing on insulin-sensitive organs (liver, skeletal muscle, adipose tissue). Results: This low-dose exposure caused significant metabolic disruptions, including hyperglycemia, hyperinsulinemia, and insulin resistance. Transcriptomic analysis of liver, skeletal muscle, and adipose tissue identified key changes in metabolic and immune pathways especially in liver. Proteomic analysis of the liver revealed mitochondrial dysfunction as a shared feature, with disruptions in oxidative phosphorylation, mitophagy, and immune activation. Comparative analysis with high-dose (50 ppm) exposure showed both conserved and dose-specific transcriptomic changes in liver, particularly in metabolic and immune responses. Conclusions: Our study is the first to comprehensively assess the impacts of occupational benzene exposure on metabolic health, highlighting mitochondrial dysfunction as a central mechanism and the dose-dependent molecular pathways in insulin-sensitive organs driving benzene-induced metabolic imbalance. Our data indicate that current OSHA occupational exposure limits for benzene are insufficient, as they could result in adverse metabolic health in exposed workers, particularly men, following chronic exposure.

pharmacology and toxicology↗

bioRxiv2024

Neurotoxic Methamphetamine Doses Alter CDCel-1 Levels and Its Interaction with Vesicular Monoamine Transporter-2 in Rat Striatum

In recent years, methamphetamine METH misuse in the US has been rapidly increasing and there is no FDA-approved pharmacotherapy for METH use disorder (MUD). In addition to being dependent on the drug, people with MUD develop a variety of neurological problems related to the toxicity of this drug. A variety of molecular mechanisms underlying METH neurotoxicity has been identified, including dysfunction of the neuroprotective protein parkin. However, it is not known whether parkin loss of function within striatal dopaminergic (DAergic) terminals translates into a decrease in DA storage capacity. This study examined the relationship between parkin, its substrate cell division cycle related-1 (CDCrel-1), and vesicular monoamine transporter-2 (VMAT2) in METH neurotoxicity in male Sprague Dawley rats. To also assess individual differences in response to METHs neurotoxic effects, a large group of rats was treated with binge METH or saline and sacrificed 1h or 24h later. This study is the first to show that binge METH alters the levels and subcellular localization of CDCrel-1 and that CDCrel-1 interacts with VMAT2 and increases its levels at the plasma membrane. Furthermore, we found wide individual differences in the responses of measured indices to METH. Proteomic analysis of VMAT-2-associated proteins revealed upregulation of several proteins involved in the exocytosis/endocytosis cycle. The results suggest that at 1h after METH binge, DAergic neurons are engaged in counteracting METH-induced toxic effects, including oxidative stress- and hyperthermia-induced inhibition of synaptic vesicle cycling, with the responses varying between individual rats. Studying CDCrel-1, VMAT2, and other proteins in large groups of outbred rats can help define individual genetic and molecular differences in responses to METH neurotoxicity which, in turn, will aid treating humans suffering from METH use disorder and its neurological consequences.

neuroscience↗

bioRxiv2020

2019 Association of Biomolecular Resource Facilities Multi-Laboratory Data-Independent Acquisition Study

Despite the advantages of fewer missing values by collecting fragment ion data on all analytes in the sample, as well as the potential for deeper coverage, the adoption of data-independent acquisition (DIA) in core facility settings has been slow. The Association of Biomolecular Resource Facilities conducted a large interlaboratory study to evaluate DIA performance in laboratories with various instrumentation. Participants were supplied with generic methods and a uniform set of test samples. The resulting 49 DIA datasets act as benchmarks and have utility in education and tool development. The sample set consisted of a tryptic HeLa digest spiked with high or low levels of four exogenous proteins. Data are available in MassIVE MSV000086479. Additionally, we demonstrate how the data can be analysed by focusing on two datasets using different library approaches and show the utility of select summary statistics. These data can be used by DIA newcomers, software developers, or DIA experts evaluating performance with different platforms, acquisition settings and skill levels.

bioinformatics↗