Biology subjects

Werner, L.

Publications and source records attributed to Werner, L..

7 recordsLinked to original sources

bioRxiv2026

A chemically reactive and Raman-active non-canonical amino acid reveals photocycle complexity in a blue-light receptor

Photosensory protein function spans multiple time and length scales, demanding integrative approaches. We introduce 4-diacetylenyl-phenylalanine (DAF), a dual-purpose non-canonical amino acid (ncAA) that enables both chemical control and spectroscopic readout of photoreceptor dynamics. Genetically encoded in E. coli, DAF combines a reactive diyne for bioorthogonal ligations (thiols, azides, tetrazines) with a strong, solvatochromic Raman signal in the cell-silent region. Applied to the light-oxygen-voltage (LOV) transcription factor EL222, DAF enables multifaceted interrogation of its photocycle. We engineer a covalently cross-linked variant that suppresses light-driven conformational changes and DNA binding, and generate a donor-acceptor construct for Forster resonance energy transfer (FRET) tracking of photoinduced structural dynamics. Time-resolved stimulated Raman spectroscopy following flavin mononucleotide (FMN) excitation reveals additional processes -from vibrational energy transfer to local unfolding-spanning femtoseconds to milliseconds. DAF thus constitutes a versatile tool to resolve protein dynamics with high spatiotemporal resolution.

biochemistry↗

bioRxiv2025

Evaluation of a Polarization-Sensitive, Dual-Wavelength Wearable Photoplethysmography Sensor Across a Range of Skin Tones

SignificanceHigh-quality photoplethysmography (PPG) signals are essential for accurate extraction of cardiovascular metrics such as heart rate, heart rate variability, and perfusion index. However, signal degradation for individuals with dark skin tones can compromise PPG quality and pose challenges for equitable sensing. AimWe develop a dual-wavelength, polarization-sensitive PPG device to assess perfusion index (PI) across a range of skin tones. ApproachTo evaluate the impact of polarization on PPG signal quality, we record PI for co-polarized (polarized illumination and parallel-aligned polarized detection), and cross-polarized conditions (polarized illumination and orthogonally aligned polarized detection) at 655 nm and 940 nm in participants representing light, medium, and brown skin tone categories. Skin tone classification are based on the individual typology angle (ITA) values derived from the CIE L*b* color space measurements. ResultsAt 940 nm, light from the cross-polarized light channel significantly increases PI (p < 0.05). At 655 nm, cross-polarization presents a statistically significantly enhanced PI (p < 0.05) relative to light from the co-polarized illumination condition, although the magnitude of the improvement decreases with lighter skin tone indication a possible interaction between skin tone and polarization. This improvement is consistent across all skin tones. ConclusionsOur results suggest that the cross-polarized condition improves PPG signal quality by reducing the influence of superficial scattering and enhancing deeper vascular signals. This approach may be especially beneficial for individuals with darker skin tones and offers a promising path towards more robust and inclusive physiological monitoring using PPG-based technologies.

bioengineering↗

bioRxiv2024

Molecular dissection of HERV-W dependent microglial- and astroglial cell polarization

Abstract/SubjectThe endogenous retrovirus type W (HERV-W) is a human-specific entity, which was initially discovered in multiple sclerosis (MS) patient derived cells. We initially found that the HERV-W envelope (ENV) protein negatively affects oligodendrogenesis and controls microglial cell polarization towards a myelinated axon associated and damaging phenotype. Such first functional assessments were conducted ex vivo, given the human-specific origin of HERV-W. Recent experimental evidence gathered on a novel transgenic mouse model, mimicking activation and expression of the HERV-W ENV protein, revealed that all glial cell types are impacted and that cellular fates, differentiation, and functions were changed. In order to identify HERV-W-specific signatures in glial cells, the current study analyzed the transcriptome of ENV protein stimulated microglial- and astroglial cells and compared the transcriptomic signatures to lipopolysaccharide (LPS) stimulated cells, owing to the fact that both ligands can activate toll-like receptor-4 (TLR-4). Additionally, a comparison between published disease associated glial signatures and the transcriptome of HERV-W ENV stimulated glial cells was conducted. We, therefore, provide here for the first time a detailed molecular description of specific HERV-W ENV evoked effects on those glial cell populations that are involved in smoldering neuroinflammatory processes relevant for progression of neurodegenerative diseases.

neuroscience↗

bioRxiv2024

Mitochondrially targeted deferasirox kills cancer cells via simultaneous iron deprivation and ferroptosis induction.

In principle, two separate ways to target cancer cells exist, the first one using iron deprivation and subsequent dysfunction of iron-dependent enzymes, and the second one inducing iron overload that leads to cell death known as ferroptosis. In this study, we introduce a compound that uniquely employs both pathways at once - mitochondrially targeted deferasirox (mitoDFX). MitoDFX deprives cells of biologically active iron while simultaneously depleting the primary cellular antioxidant glutathione (GSH) and inducing lipid peroxidation, both of which are hallmarks of ferroptosis. The role of the GSH is further supported by enhanced cell death in glutathione peroxidase 4 KO cells (GPX4 KO) induced by mitoDFX. This response is further exacerbated by simultaneous inhibition of glutathione metabolism or the pentose phosphate pathway. MitoDFX strongly affects the structure and function of mitochondria, leading to its fragmentation, ROS production and induction of mitophagy. Moreover, we found that mitoDFX treatment markedly reduces mitochondrial translation and downregulates levels of enzymatic subunits coupled with the ETC/TCA cycle as well as DNA replication and transcription, which could explain its strong cytostatic effects. In summary, mitoDFX achieves high activity and selectivity against cancer cells that seems unattainable for the conventional untargeted iron chelators, which have adverse effects on systemic iron metabolism. Our novel, targeted chelator has also shown enhanced efficacy in animal models, surpassing other mitochondrially targeted drugs and making mitoDFX a promising candidate for the development of future selective and highly effective cancer therapies.

cancer biology↗

bioRxiv2022

Repurposing of mitochondria-targeted tamoxifen: Novel anti-cancer drug exhibits potent activity against major protozoan and fungal pathogens.

Many of the currently available anti-parasitic and anti-fungal frontline drugs have severe limitations, including adverse side effects, complex administration, and increasing occurrence of resistance. The discovery and development of new therapeutic agents is a costly and lengthy process. Therefore, repurposing drugs with already established clinical application offers an attractive, fast-track approach for novel treatment options. In this study, we show that the anti-cancer drug MitoTam, a mitochondria-targeted analog of tamoxifen, efficiently eliminates a wide range of evolutionarily distinct pathogens in vitro, including pathogenic fungi, Plasmodium falciparum, and several species of trypanosomatid parasites, causative agents of debilitating neglected tropical diseases. MitoTam treatment was also effective in vivo and significantly reduced parasitemia of two medically important parasites, Leishmania mexicana and Trypanosoma brucei, in their respective animal infection models. Functional analysis in the bloodstream form of T. brucei showed that MitoTam rapidly altered mitochondrial functions, particularly affecting cellular respiration, lowering ATP levels, and dissipating mitochondrial membrane potential. Our data suggest that the mode of action of MitoTam involves disruption of the inner mitochondrial membrane, leading to rapid organelle depolarization and cell death. Altogether, MitoTam is an excellent candidate drug against several important pathogens, for which there are no efficient therapies and for which drug development is not a priority. Author SummaryMitoTam, a mitochondrially targeted analog of tamoxifen, is a promising anti-cancer candidate drug acting by accumulating in and destabilizing cell mitochondria. In this study, we analyze its effect on a wide range of evolutionarily distinct and medically important pathogens. These include a) pathogenic fungi, Candida albicans, and Cryptococcus neoformans, ubiquitous opportunistic pathogens that cause life-threatening diseases in immunocompromised or immunologically deficient individuals; b) Plasmodium falciparum, the causative agent of human malaria; and c) several species of trypanosomatid parasites such as Trypanosoma cruzi, responsible for deadly Chagas disease in South America, Trypanosoma brucei, the causative agent of sleeping sickness in Africa, and Leishmania, the etiological agent of leishmaniasis, a spectrum of diseases ranging from usually self-healing but potentially disfiguring cutaneous and mucocutaneous leishmaniasis, to visceral leishmaniasis, which is invariably fatal if left untreated. We show that MitoTam efficiently kills these parasites in laboratory conditions and in the case of trypanosomes and leishmaniases suppress or at least slow down the infection in mouse model.

microbiology↗

bioRxiv2022

Single cell atlas of the neonatal small intestine with necrotizing enterocolitis

Necrotizing enterocolitis (NEC) is a gastrointestinal complication of premature infants with high rates of morbidity and mortality. A comprehensive view of the cellular changes and aberrant interactions that underlie this disease is lacking. Here, we combine single cell RNA sequencing, T Cell Receptor beta (TCR{beta}) analysis, bulk transcriptomics, and imaging to characterize cell identities, interactions and zonal changes in NEC. We find that inflammatory macrophages are abundant in NEC and that T cells exhibit increased expression of inflammatory genes and cytokines accompanied by an increase in TCR{beta} clonal expansion. Fibroblasts and endothelial cells increase in proportion and exhibit a switch to an activated pro-inflammatory state. Villus tip epithelial cell identity is substantially reduced in NEC and the remaining epithelial cells up-regulate pro-inflammatory genes. We establish a detailed map of aberrant epithelial-mesenchymal-immune interactions that may be driving inflammation in NEC mucosa. Our analyses highlight the cellular changes underlying NEC disease pathogenesis and identify potential targets for biomarker discovery and therapeutics.

immunology↗

bioRxiv2019

Multidimensional extracellular-matrix analysis reveals a silent pre-clinical signature in inflammatory disease

Identification of early processes leading to complex tissue pathologies, such as inflammatory bowel diseases, poses a major scientific and clinical challenge that is imperative for improved diagnosis and treatment. Most studies of inflammation onset focus on cellular processes and signaling molecules, while overlooking the environment in which they take place, the continuously remodeled extracellular matrix. In this study, we used colitis models for investigating extracellular-matrix dynamics during disease onset, while treating the matrix as a complete and defined entity. Through the analysis of matrix structure, stiffness and composition, we unexpectedly revealed that even prior to the first clinical symptoms, the colon displays its own unique extracellular-matrix signature and found specific markers of clinical potential, which were also validated in human subjects. We also show that the emergence of this pre-symptomatic matrix is mediated by sub-clinical infiltration of neutrophils and monocytes bearing remodeling enzymes. Remarkably, whether the inflammation is chronic or acute, its matrix signature converges at pre-symptomatic states. We suggest that the existence of a pre-symptomatic extracellular-matrix is general and relevant to a wide range of diseases.

systems biology↗