Ghyslain Budin - Academia.edu (original) (raw)

Papers by Ghyslain Budin

Optics and Biophotonics in Low-Resource Settings VIII

The invention relates to a technique to detect small molecules using Bioluminescence imaging (BLI... more The invention relates to a technique to detect small molecules using Bioluminescence imaging (BLI) to image and quantify non-invasively, in vitro and in vivo,intracellular metabolite fluxes and which can be applied to azido-modified compounds, such as azido-modified biomolecules

L’objectif majeur de ce travail a été de développer une boite à outil moléculaire permettant la v... more L’objectif majeur de ce travail a été de développer une boite à outil moléculaire permettant la visualisation des protéines dans la cellule et leur identification par spectrométrie de masse. Ces outils ont été conçus pour réagir de façon bioorthogonale avec des sondes chimiques ciblant une classe spécifique de protéine. Le premier objectif a été de développer une approche de marquage fluorescent des protéines en conditions non-covalentes et non-dénaturantes puis de développer un système pour la purification de complexe protéique en conditions non-dénaturantes pour identifier les sous-unités protéiques.The general topic of this work was to develop new chemicals and biochemical tools box allowing proteinsvisualization in cells and identification of proteins by mass spectrometry. These tools were designed to react bioorthogonally with chemicals probes that target a specific class of protein. The first objective was to develop a non-covalent and non-denaturing fluorescent labeling appro...

David Díaz Díaz,* Emmanuelle Morin, Eva M. Schön, Ghyslain Budin, Alain Wagner and JeanSerge Remy... more David Díaz Díaz,* Emmanuelle Morin, Eva M. Schön, Ghyslain Budin, Alain Wagner and JeanSerge Remy a Institut für Organische Chemie, Universität Regensburg, Universitätsstr. 31, 93040 Regensburg, Germany b Instituto de Ciencias de Materiales de Aragón, CSIC-Universidad de Zaragoza, Pedro Cerbuna 12, 50009 Zaragoza, Spain c Chimie Génétique, Faculté de Pharmacie, 67000 Strasbourg, France d Laboratory of Functionnal ChemoSystems, UMR 7199, Faculté de Pharmacie, 67000 Strasbourg, France

Nature Communications, 2021

Bioluminescent imaging (BLI) is one of the most powerful and widely used preclinical imaging moda... more Bioluminescent imaging (BLI) is one of the most powerful and widely used preclinical imaging modalities. However, the current technology relies on the use of transgenic luciferase-expressing cells and animals and therefore can only be applied to a limited number of existing animal models of human disease. Here, we report the development of a “portable bioluminescent” (PBL) technology that overcomes most of the major limitations of traditional BLI. We demonstrate that the PBL method is capable of noninvasive measuring the activity of both extracellular (e.g., dipeptidyl peptidase 4) and intracellular (e.g., cytochrome P450) enzymes in vivo in non-luciferase-expressing mice. Moreover, we successfully utilize PBL technology in dogs and human cadaver, paving the way for the translation of functional BLI to the noninvasive quantification of biological processes in large animals. The PBL methodology can be easily adapted for the noninvasive monitoring of a plethora of diseases across mult...

L’objectif majeur de ce travail a ete de developper une boite a outil moleculaire permettant la v... more L’objectif majeur de ce travail a ete de developper une boite a outil moleculaire permettant la visualisation des proteines dans la cellule et leur identification par spectrometrie de masse. Ces outils ont ete concus pour reagir de facon bioorthogonale avec des sondes chimiques ciblant une classe specifique de proteine. Le premier objectif a ete de developper une approche de marquage fluorescent des proteines en conditions non-covalentes et non-denaturantes puis de developper un systeme pour la purification de complexe proteique en conditions non-denaturantes pour identifier les sous-unites proteiques.

Nature Chemical Biology

Mitochondrial membrane potential (Δ Ψ m ) is a universal selective indicator of mitochondrial fun... more Mitochondrial membrane potential (Δ Ψ m ) is a universal selective indicator of mitochondrial function and is known to play a central role in many human pathologies, such as diabetes mellitus, cancer and Alzheimer’s and Parkinson’s diseases. Here, we report the design, synthesis and several applications of mitochondria-activatable luciferin (MAL), a bioluminescent probe sensitive to Δ Ψ m , and partially to plasma membrane potential (Δ Ψ p ), for non-invasive, longitudinal monitoring of Δ Ψ m in vitro and in vivo. We applied this new technology to evaluate the aging-related change of Δ Ψ m in mice and showed that nicotinamide riboside (NR) reverts aging-related mitochondrial depolarization, revealing another important aspect of the mechanism of action of this potent biomolecule. In addition, we demonstrated application of the MAL probe for studies of brown adipose tissue (BAT) activation and non-invasive in vivo assessment of Δ Ψ m in animal cancer models, opening exciting opportunities for understanding the underlying mechanisms and for discovery of effective treatments for many human pathologies. A mitochondria-activatable bioluminescent probe was designed enabling sensitive, non-invasive and longitudinal monitoring of mitochondrial membrane potential in vitro and in vivo.

The Journal of the American Academy of Orthopaedic Surgeons, Jan 8, 2016

Diagnosing prosthetic joint infection (PJI) poses significant challenges, and current modalities ... more Diagnosing prosthetic joint infection (PJI) poses significant challenges, and current modalities are fraught with low sensitivity and/or potential morbidity. Photoacoustic imaging (PAI) is a novel ultrasound-based modality with potential for diagnosing PJI safely and noninvasively. In an established preclinical mouse model of bioluminescent Staphylococcus aureus PJI, fluorescent indocyanine green (ICG) was conjugated to β-cyclodextrin (CDX-ICG) or teicoplanin (Teic-ICG) and injected intravenously for 1 week postoperatively. Daily fluorescent imaging and PAI were used to localize and quantify tracer signals. Results were analyzed using 2-way analysis of variance. Fluorescence clearly localized to the site of infection and was significantly higher with Teic-ICG compared with CDX-ICG (P = 0.046) and ICG alone (P = 0.0087). With PAI, the photoacoustic signal per volumetric analysis was substantially higher and better visualized with Teic-ICG compared with CDX-ICG and ICG alone, and colo...

Advanced Healthcare Materials, 2013

Sensitive, rapid and phenotype-specific enumeration of pathogens is essential for the diagnosis o... more Sensitive, rapid and phenotype-specific enumeration of pathogens is essential for the diagnosis of infectious disease, monitoring of food chains, and for defense against bioterrorism. Microbiological culture and genotyping, techniques that sensitively and selectively detect bacteria in laboratory settings, have limited application in clinical environments due to high cost, slow response times, and the need for specially trained staff and laboratory infrastructure. To address these challenges, we developed a microfluidic chip-based micro-Hall (μHall) platform capable of measuring single, magnetically tagged bacteria directly in clinical specimens with minimal sample processing. We demonstrated the clinical utility of the μHall chip by enumerating Gram-positive bacteria using a two-step bioorthogonal labeling procedure. The overall detection limit of the system was similar to that of culture tests (~10 bacteria), but the assay time was 50-times faster. This low-cost, single-cell analytical technique is especially well-suited to diagnose infectious diseases in resource-limited clinical settings. Keywords μHall; bacterial detection; magnetic nanoparticles Development of fast, sensitive, and accurate diagnostic tests is key to controlling infectious diseases. [1-3] By enabling early diagnosis and treatment, such tests would not only help rapidly identify sources of infection, but would prevent further disease transmission, and reduce the risk of patients developing long-term complications. [2] The detection of pathogens (bacteria, parasites, fungi, viruses) in clinical specimens, however, has continued to present significant technical challenges, namely because a) specimens often have a complex composition (e.g., blood, tissue), b) sample sizes are often limited, and c) specimens may contain very few pathogens (e.g., early disease stages). [4-7] Magnetic sensing, a technique in which pathogens are labeled with magnetic nanoparticles (MNPs), is

Current Protocols in Chemical Biology, 2009

The great complexity of many human pathologies, such as cancer, diabetes, and neurodegenerative d... more The great complexity of many human pathologies, such as cancer, diabetes, and neurodegenerative diseases, requires new tools for studies of biological processes on the whole organism level. The discovery of novel biocompatible reactions has tremendously advanced our understanding of basic biology; however, no efficient tools exist for realtime non-invasive imaging of many human proteases that play very important roles in multiple human disorders. We recently reported that the "split luciferin" biocompatible reaction represents a valuable tool for evaluation of protease activity directly in living animals using bioluminescence imaging (BLI). Since BLI is the most sensitive in vivo imaging modality known to date, this method can be widely applied for the evaluation of the activity of multiple proteases, as well as identification of their new peptide-specific substrates. In this unit, we describe several applications of this "split luciferin" reaction for quantification of protease activities in test tube assays and living animals. Curr.

Tetrahedron Lett, 2009

The direct azidation of various heterocyclic β-ketoesters, lactones, and lactams is reported. By ... more The direct azidation of various heterocyclic β-ketoesters, lactones, and lactams is reported. By using tosylazide and an organic base such as l-proline or TBD, the direct α-insertion of azide into these substrates was achieved in moderate to good yields, without competitive deacylating diazo transfer. This procedure represents an interesting alternative to the usual two-step approach of α-halogenation and subsequent displacement with azide ion.

PLOS ONE, 2015

Trehalose is a naturally occurring disaccharide which is associated with extraordinary stress-tol... more Trehalose is a naturally occurring disaccharide which is associated with extraordinary stress-tolerance capacity in certain species of unicellular and multicellular organisms. In mammalian cells, presence of intra- and extracellular trehalose has been shown to confer improved tolerance against freezing and desiccation. Since mammalian cells do not synthesize nor import trehalose, the development of novel methods for efficient intracellular delivery of trehalose has been an ongoing investigation. Herein, we studied the membrane permeability of engineered lipophilic derivatives of trehalose. Trehalose conjugated with 6 acetyl groups (trehalose hexaacetate or 6-O-Ac-Tre) demonstrated superior permeability in rat hepatocytes compared with regular trehalose, trehalose diacetate (2-O-Ac-Tre) and trehalose tetraacetate (4-O-Ac-Tre). Once in the cell, intracellular esterases hydrolyzed the 6-O-Ac-Tre molecules, releasing free trehalose into the cytoplasm. The total concentration of intracellular trehalose (plus acetylated variants) reached as high as 10 fold the extracellular concentration of 6-O-Ac-Tre, attaining concentrations suitable for applications in biopreservation. To describe this accumulation phenomenon, a diffusion-reaction model was proposed and the permeability and reaction kinetics of 6-O-Ac-Tre were determined by fitting to experimental data. Further studies suggested that the impact of the loading and the presence of intracellular trehalose on cellular viability and function were negligible. Engineering of trehalose chemical structure rather than manipulating the cell, is an innocuous, cell-friendly method for trehalose delivery, with demonstrated potential for trehalose loading in different types of cells and cell lines, and can facilitate the wide-spread application of trehalose as an intracellular protective agent in biopreservation studies.

Tetrahedron Letters, 2009

European Journal of Organic Chemistry, 2010

ABSTRACT In this paper we conducted an extensive reactivity study to determine the key structural... more ABSTRACT In this paper we conducted an extensive reactivity study to determine the key structural features that favour the dithionite-triggered reductive cleavage of the azo–arene group. Our stepwise investigation allowed identification of a highly reactive azo–arene structure 25 bearing a carboxylic acid at the ortho position of the electron-poor arene and an ortho-O-alkyl-resorcinol as the electron-rich arene. Based on this 2-(2′-alkoxy-4′-hydroxyphenylazo)benzoic acid (HAZA) scaffold, the orthogonally protected difunctional azo–arene cleavable linker 26 was designed and synthesized. Selective linker deprotection and derivatization was performed by introducing an alkyne reactive group and a biotin affinity tag. This optimized azo–arene cleavable linker led to a total cleavage in less than 10 s with only 1 mM dithionite. Similar results were obtained in biological media.