Glycaemic control in diabetic rats treated with islet transplantation using plasma combined with hydroxypropylmethyl cellulose hydrogel

Islet transplantation is one of the most efficient cell therapies used in clinics and could treat a large proportion of patients with diabetes. However, it is limited by the high requirement of pancreas necessary to provide the sufficient surviving islet mass in the hepatic tissue and restore normoglycaemia. Reduction in organ procurement requirements could be achieved by extrahepatic transplantation using a biomaterial that enhances islet survival and function. We report a plasma-supplemented hydroxypropyl methylcellulose (HPMC) hydrogel, engineered specifically using a newly developed technique for intra-omental islet infusion, known as hOMING (h-Omental Matrix Islet filliNG). The HPMC hydrogel delivered islets with better performance than that of the classical intrahepatic infusion. After the validation of the HPMC suitability for islets in vivo and in vitro, plasma supplementation modified the rheological properties of HPMC without affecting its applicability with hOMING. The biomaterial association was proven to be more efficient both in vitro and in vivo, with better islet viability and function than that of the current clinical intrahepatic delivery technique. Indeed, when the islet mass was decreased by 25% or 35%, glycaemia control was observed in the group of plasma-supplemented hydrogels, whereas no regulation was observed in the hepatic group. Plasma gelation, observed immediately post infusion, decreased anoïkis and promoted vascularisation. To conclude, the threshold mass for islet transplantation could be decreased using HPMC-Plasma combined with the hOMING technique. The simplicity of the hOMING technique and the already validated use of its components could facilitate its transfer to clinics

Acétylation des histones au cours des processus de mémorisation : influence du vieillissement et de l'environnement enrichi

Hippocampal-dependent memory formation is associated with the establishment of specific genetic programs in the rat hippocampus. This transcriptional activation of genes involved in synaptic plasticity and memory processes, like bdnf, can in part be attributable to histone acetylation-related mechanisms, allowing dynamic chromatin structure changes. Our results indicate a specific and differential regulation of histone acetylation in young rodents hippocampus depending on the nature of the stimuli. In fact, H2B and H4 acetylations are specific to rats having learnt a task (MWM, CFC), whereas H3 acetylation seems to be more sensitive to the environmental context. Besides, it is known that ageing and environmental enrichment (EE) are factors able to modulate histone acetylation, leading respectively to repression and activation of memory-related genetic programs. Here, we showed that an EE of 6 months, even applied to 18 month-old rats, which already present memory deficits, is able to induce persistent chromatin structure modifications through H3. By favoring the expression of genes as bdnf, these changes could participate in the preservation of memory abilities, which are normally lost in 24 month-old rats. The precise identification of regulating elements located on the bdnf promoter brings new data about the potential factors involved in the transcriptional response following EE, e.g. CREB and NFκB. Altogether, our results confirm the role of histone acetylation in memory processes and underline that these regulations remain flexible during life, thus highlighting possible therapeutic strategies in pathological ageing conditions.La formation de souvenirs nécessite la mise en place de programmes génétiques dans l’hippocampe. L’activation de la transcription de gènes impliqués dans les processus de plasticité comme le bdnf s’effectue, au moins en partie, via l’acétylation des histones, mécanisme qui permet des changements de la structure de la chromatine. Nos résultats soulignent l’existence d’une régulation spécifique et différentielle de l’acétylation des histones dans l’hippocampe de rongeurs adultes en fonction du type d’information à traiter. Les acétylations des histones H2B et H4 sont spécifiques de l’apprentissage d’une tâche (MWM, CFC) alors que celle de l’histone H3 semble plus sensible au contexte environnemental. Il est par ailleurs décrit que le vieillissement ainsi que l’environnement enrichi (EE) sont des facteurs susceptibles d’induire des changements d’acétylation des histones, aboutissant respectivement à la répression et à l’activation de gènes de « mémoire ». Nos études mettent en évidence qu’un EE de 6 mois, même appliqué à des rats âgés de 18 mois qui présentent déjà des déficits mnésiques, est capable d’induire des modifications durables de la structure de la chromatine par l’intermédiaire de H3. En favorisant l’expression de gènes comme le bdnf, ces changements participent au maintien des capacités mnésiques, normalement perdues chez le Rat âgé de 24 mois. Dans l’ensemble, nos résultats soulignent l’importance des mécanismes liés aux acétylations des histones dans les processus mnésiques et indiquent que ces régulations restent modulables au cours de la vie, permettant d’envisager d’éventuelles options thérapeutiques dans des conditions de vieillissement pathologique

Histone acetylation and memory processes : impact of ageing and environmental enrichment

La formation de souvenirs nécessite la mise en place de programmes génétiques dans l’hippocampe. L’activation de la transcription de gènes impliqués dans les processus de plasticité comme le bdnf s’effectue, au moins en partie, via l’acétylation des histones, mécanisme qui permet des changements de la structure de la chromatine. Nos résultats soulignent l’existence d’une régulation spécifique et différentielle de l’acétylation des histones dans l’hippocampe de rongeurs adultes en fonction du type d’information à traiter. Les acétylations des histones H2B et H4 sont spécifiques de l’apprentissage d’une tâche (MWM, CFC) alors que celle de l’histone H3 semble plus sensible au contexte environnemental. Il est par ailleurs décrit que le vieillissement ainsi que l’environnement enrichi (EE) sont des facteurs susceptibles d’induire des changements d’acétylation des histones, aboutissant respectivement à la répression et à l’activation de gènes de « mémoire ». Nos études mettent en évidence qu’un EE de 6 mois, même appliqué à des rats âgés de 18 mois qui présentent déjà des déficits mnésiques, est capable d’induire des modifications durables de la structure de la chromatine par l’intermédiaire de H3. En favorisant l’expression de gènes comme le bdnf, ces changements participent au maintien des capacités mnésiques, normalement perdues chez le Rat âgé de 24 mois. Dans l’ensemble, nos résultats soulignent l’importance des mécanismes liés aux acétylations des histones dans les processus mnésiques et indiquent que ces régulations restent modulables au cours de la vie, permettant d’envisager d’éventuelles options thérapeutiques dans des conditions de vieillissement pathologique.Hippocampal-dependent memory formation is associated with the establishment of specific genetic programs in the rat hippocampus. This transcriptional activation of genes involved in synaptic plasticity and memory processes, like bdnf, can in part be attributable to histone acetylation-related mechanisms, allowing dynamic chromatin structure changes. Our results indicate a specific and differential regulation of histone acetylation in young rodents hippocampus depending on the nature of the stimuli. In fact, H2B and H4 acetylations are specific to rats having learnt a task (MWM, CFC), whereas H3 acetylation seems to be more sensitive to the environmental context. Besides, it is known that ageing and environmental enrichment (EE) are factors able to modulate histone acetylation, leading respectively to repression and activation of memory-related genetic programs. Here, we showed that an EE of 6 months, even applied to 18 month-old rats, which already present memory deficits, is able to induce persistent chromatin structure modifications through H3. By favoring the expression of genes as bdnf, these changes could participate in the preservation of memory abilities, which are normally lost in 24 month-old rats. The precise identification of regulating elements located on the bdnf promoter brings new data about the potential factors involved in the transcriptional response following EE, e.g. CREB and NFκB. Altogether, our results confirm the role of histone acetylation in memory processes and underline that these regulations remain flexible during life, thus highlighting possible therapeutic strategies in pathological ageing conditions

Acétylation des histones au cours des processus de mémorisation (influence du vieillissement et de l'environnement enrichi)

La formation de souvenirs nécessite la mise en place de programmes génétiques dans l hippocampe. L activation de la transcription de gènes impliqués dans les processus de plasticité comme le bdnf s effectue, au moins en partie, via l acétylation des histones, mécanisme qui permet des changements de la structure de la chromatine. Nos résultats soulignent l existence d une régulation spécifique et différentielle de l acétylation des histones dans l hippocampe de rongeurs adultes en fonction du type d information à traiter. Les acétylations des histones H2B et H4 sont spécifiques de l apprentissage d une tâche (MWM, CFC) alors que celle de l histone H3 semble plus sensible au contexte environnemental. Il est par ailleurs décrit que le vieillissement ainsi que l environnement enrichi (EE) sont des facteurs susceptibles d induire des changements d acétylation des histones, aboutissant respectivement à la répression et à l activation de gènes de mémoire . Nos études mettent en évidence qu un EE de 6 mois, même appliqué à des rats âgés de 18 mois qui présentent déjà des déficits mnésiques, est capable d induire des modifications durables de la structure de la chromatine par l intermédiaire de H3. En favorisant l expression de gènes comme le bdnf, ces changements participent au maintien des capacités mnésiques, normalement perdues chez le Rat âgé de 24 mois. Dans l ensemble, nos résultats soulignent l importance des mécanismes liés aux acétylations des histones dans les processus mnésiques et indiquent que ces régulations restent modulables au cours de la vie, permettant d envisager d éventuelles options thérapeutiques dans des conditions de vieillissement pathologique.Hippocampal-dependent memory formation is associated with the establishment of specific genetic programs in the rat hippocampus. This transcriptional activation of genes involved in synaptic plasticity and memory processes, like bdnf, can in part be attributable to histone acetylation-related mechanisms, allowing dynamic chromatin structure changes. Our results indicate a specific and differential regulation of histone acetylation in young rodents hippocampus depending on the nature of the stimuli. In fact, H2B and H4 acetylations are specific to rats having learnt a task (MWM, CFC), whereas H3 acetylation seems to be more sensitive to the environmental context. Besides, it is known that ageing and environmental enrichment (EE) are factors able to modulate histone acetylation, leading respectively to repression and activation of memory-related genetic programs. Here, we showed that an EE of 6 months, even applied to 18 month-old rats, which already present memory deficits, is able to induce persistent chromatin structure modifications through H3. By favoring the expression of genes as bdnf, these changes could participate in the preservation of memory abilities, which are normally lost in 24 month-old rats. The precise identification of regulating elements located on the bdnf promoter brings new data about the potential factors involved in the transcriptional response following EE, e.g. CREB and NF B. Altogether, our results confirm the role of histone acetylation in memory processes and underline that these regulations remain flexible during life, thus highlighting possible therapeutic strategies in pathological ageing conditions.STRASBOURG-Bib.electronique 063 (674829902) / SudocSudocFranceF

Detection of Histone Acetylation Levels in the Dorsal Hippocampus Reveals Early Tagging on Specific Residues of H2B and H4 Histones in Response to Learning

International audienc

Oral insulin delivery, the challenge to increase insulin bioavailability: Influence of surface charge in nanoparticle system

International audienceOral administration of insulin increases patient comfort and could improve glycemic control thanks to the hepatic first passage. However, challenges remain. The current approach uses poly (d, lactic-co-glycolic) acid (PLGA) nanoparticles (NPs), an effective drug carrier system with a long acting profile. However, this system presents a bioavailability of less than 20% for insulin encapsulation. In this context, physico-chemical parameters like surface charge could play a critical role in NP uptake by the intestinal barrier. Therefore, we developed a simple method to modulate NP surface charge to test its impact on uptake in vitro and finally on NP efficiency in vivo. Various NPs were prepared in the presence (+) or absence (−) of polyvinyl alcohol (PVA), sodium dodecyl sulfate (SDS), and/or coated with chitosan chloride. In vitro internalization was tested using epithelial culture of Caco-2 or using a co-culture (Caco-2/RevHT29MTX) by flow cytometry. NPs were then administered by oral route using a pharmaceutical complex vector (100 or 250 UI/kg) in a diabetic rat model.SDS-NPs (−42 ± 2 mV) were more negatively charged than −PVA-NPs (–22 ± 1 mV) and chitosan-coated NPs were highly positively charged (56 ± 2 mV) compared to +PVA particles (−2 ± 1 mV), which were uncharged. In the Caco-2 model, NP internalization was significantly improved by using negatively charged NPs (SDS NPs) compared to using classical NPs (+PVA NPs) and chitosan-coated NPs. Finally, the efficacy of insulin SDS-NPs was demonstrated in vivo (100 or 250 UI insulin/kg) with a reduction of blood glucose levels in diabetic rats. Formulation of negatively charged NPs represents a promising approach to improve NP uptake and insulin bioavailability for oral delivery

Impact of contextual fear conditioning on histone acetylation in the rat hippocampus.

<p>(A) Experimental design. Three groups of rats (n = 16/group) were used. In one group, rats were kept in the context but received no shock (CX). Others received three immediate and consecutive shocks and were subsequently left in the context for 8 min (IS). In the last group, rats received three randomly-distributed shocks while being kept in the context as noted (CS). Animals (n = 10/group) were then either tested for freezing behavior after 24 h (probe) (B; n = 10/group<b>)</b> or euthanized after 1 h for tissue collection (dorsal hippocampus) and western blot analyses of acetylated histones (C; n = 6/group<b>)</b>. (B) Freezing levels at 24 h. Notice that marked freezing was observed only in the Context-shock group (CS), demonstrating that rats of this group were the only ones to have associated the shock with the context and memorized this association. (C) Comparison of acetylated and total histone levels in the three groups relative to their counterparts taken from the home cage (HC, n = 6). Lysine acetylations measured were H2BK5 (K5Ac, plain histograms), H2BK5K12K15K20 (Tetra Ac, stripped histograms), H4K12 (K12Ac) and H3K9K14 (K9K14Ac). Typical western blots are shown in duplicates. Quantified results are represented as % induction of the Acetylated/total ratio for each histone. The ratio obtained in the HC condition was arbitrarily set at 100%. Newman-Keuls multiple comparisons test: ***p<0.001**p<0.01, *p<0.05, as compared to HC group. Global H2B and H4 histone acetylation levels were clearly increased in the group exhibiting fear towards the context (CS) as compared to the other situations, while H3 acetylation levels were increased in CS and both controls (CX and IS ) as compared to rats completely naive to the test situation (HC).</p

J. Neurosci.

Aging weakens memory functions. Exposing healthy rodents or pathological rodent models to environmental enrichment (EE) housing improves their cognitive functions by changing neuronal levels of excitation, cellular signaling, and plasticity, notably in the hippocampus. At the molecular level, brain derived-neurotrophic factor (BDNF) represents an important player that supports EE-associated changes. EE facilitation of learning was also shown to correlate with chromatin acetylation in the hippocampus. It is not known, however, whether such mechanisms are still into play during aging. In this study, we exposed a cohort of aged rats (18-month-old) to either a 6 month period of EE or standard housing conditions and investigated chromatin acetylation-associated events [histone acetyltranferase activity, gene expression, and histone 3 (H3) acetylation] and epigenetic modulation of the Bdnf gene under rest conditions and during learning. We show that EE leads to upregulation of acetylation-dependent mechanisms in aged rats, whether at rest or following a learning challenge. We found an increased expression of Bdnf through Exon-I-dependent transcription, associated with an enrichment of acetylated H3 at several sites of Bdnf promoter I, more particularly on a proximal nuclear factor κB (NF-κB) site under learning conditions. We further evidenced p65/NF-κB binding to chromatin at promoters of genes important for plasticity and hippocampus-dependent learning (e.g., Bdnf, CamK2D). Altogether, our findings demonstrate that aged rats respond to a belated period of EE by increasing hippocampal plasticity, together with activating sustained acetylation-associated mechanisms recruiting NF-κB and promoting related gene transcription. These responses are likely to trigger beneficial effects associated with EE during aging. SIGNIFICANCE STATEMENT: Aging weakens memory functions. Optimizing the neuronal circuitry required for normal brain function can be achieved by increasing sensory, motor, and cognitive stimuli resulting from interactions with the environment (behavioral therapy). This can be experimentally modeled by exposing rodents to environmental enrichment (EE), as with large cages, numerous and varied toys, and interaction with other rodents. However, EE effects in aged rodents has been poorly studied, and it is not known whether beneficial mechanisms evidenced in the young adults can still be recruited during aging. Our study shows that aged rats respond to a belated period of EE by activating specific epigenetic and transcriptional signaling that promotes gene expression likely to facilitate plasticity and learning behaviors