In vivo imaging of induction of heat-shock protein-70 gene expression with fluorescence reflectance imaging and intravital confocal microscopy following brain ischaemia in reporter mice

Rosa, Xavier de la; Santalucı́a, Tomàs; Fortin, Pierre-Yves; Purroy, Jesús; Calvo, Marı́a; Salas-Perdomo, Angélica; Justicia, Carles; Couillaud, Franck; Planas, Anna M.

doi:10.1007/s00259-012-2277-7

Cited by 15 publications

(15 citation statements)

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“…Similar results of local expression of an alarmin family member could be demonstrated using a transgene reporter mice model for HSP-70 expression in ischemic brain injury. However, such transgenic approaches unfortunately only have limited potential for translation into clinical applications36. Owing to their specific mode of expression and release, S100A8 and S100A9 are sensitive biomarkers for the immediate response of innate immune mechanisms to disturbances of tissue homeostasis.…”

Section: Discussionmentioning

confidence: 99%

Alarmin S100A8/S100A9 as a biomarker for molecular imaging of local inflammatory activity

et al. 2014

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Inflammation has a key role in the pathogenesis of various human diseases. The early detection, localization and monitoring of inflammation are crucial for tailoring individual therapies. However, reliable biomarkers to detect local inflammatory activities and to predict disease outcome are still missing. Alarmins, which are locally released during cellular stress, are early amplifiers of inflammation. Here, using optical molecular imaging, we demonstrate that the alarmin S100A8/S100A9 serves as a sensitive local and systemic marker for the detection of even sub-clinical disease activity in inflammatory and immunological processes like irritative and allergic contact dermatitis. In a model of collagen-induced arthritis, we use S100A8/S100A9 imaging to predict the development of disease activity. Furthermore, S100A8/S100A9 can act as a very early and sensitive biomarker in experimental leishmaniasis for phagocyte activation linked to an effective Th1-response. In conclusion, the alarmin S100A8/S100A9 is a valuable and sensitive molecular target for novel imaging approaches to monitor clinically relevant inflammatory disorders on a molecular level.

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Section: Discussionmentioning

confidence: 99%

Alarmin S100A8/S100A9 as a biomarker for molecular imaging of local inflammatory activity

et al. 2014

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“…Some molecules may exhibit different expression pattern in different ischemic stroke model. For example, heat shock protein 70 was induced following the transient focal cerebral ischemia model,33, 34, 35 but was equally expressed in both peri‐infarct and healthy tissues in the permanent focal cerebral ischemia model36; sirtuin 1 expression was increased in neurons of the ipsilesional mouse brain in transient MCAO model,37 but was decreased in the ischemic hemisphere in the permanent MCAO model 38. To study a candidate for molecular therapeutic or imaging target in a variety of cerebral ischemic stroke may provide adequate information for its application.…”

Section: Discussionmentioning

confidence: 99%

Neuronal PirB Upregulated in Cerebral Ischemia Acts as an Attractive Theranostic Target for Ischemic Stroke

Wang

Zhang

Xia

et al. 2018

JAHA

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BackgroundIschemic stroke is a complex disease with multiple etiologies and clinical manifestations. Paired immunoglobulin‐like receptor B (PirB), which is originally thought to function exclusively in the immune system, is now also known to be expressed by neurons. A growing number of studies indicate that PirB can inhibit neurite outgrowth and restrict neuronal plasticity. The aim of the study is to investigate whether PirB can be an attractive theranostic target for ischemic stroke.Methods and ResultsFirst, we investigated the spatial‐temporal expression of PirB in multiple ischemic stroke models, including transient middle cerebral artery occlusion, photothrombotic cerebral cortex ischemia, and the neuronal oxygen glucose deprivation model. Then, anti‐PirB immunoliposome nanoprobe was developed by thin‐film hydration method and investigated its specific targeting in vitro and in vivo. Finally, soluble PirB ectodomain (sPirB) protein delivered by polyethylene glycol–modified nanoliposome was used as a therapeutic reagent for ischemic stroke by blocking PirB binding to its endogenous ligands. These results showed that PirB was significantly upregulated after cerebral ischemic injury in ischemic stroke models. Anti‐PirB immunoliposome nanoprobe was successfully developed and specifically bound to PirB in vitro. There was accumulation of anti‐PirB immunoliposome nanoprobe in the ischemic hemisphere in vivo. Soluble PirB ectodomains remarkably improved ischemic stroke model recovery by liposomal delivery system.ConclusionsThese data indicated that PirB was a significant element in the pathological process of cerebral ischemia. Therefore, PirB may act as a novel theranostic target for ischemic stroke.

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“…The inserts (italics) were PCR-amplified from the following plasmid templates: mKate2 , pmKate2-C (Evrogen); EGFP , pLV-EGFP (kind gift from M. Perez-Pinzon, UM); EYFP , pEYFP-mito (Clontech); ECFP and IRES_ECFP , pYIC (Addgene plasmid 18673 [43]; V5-6xHis epitope tag cassette , pEF6/V5-His (Life Technologies). Construction of the clone containing the SV40 early polyadenylation signal has previously been described [44]. All inserts were amplified in a two-step PCR reaction, similarly to the way described in the section above, with the primers described in Additional file 3: Table S1.…”

Section: Methodsmentioning

confidence: 99%

A plasmid toolkit for cloning chimeric cDNAs encoding customized fusion proteins into any Gateway destination expression vector

et al. 2013

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BackgroundValuable clone collections encoding the complete ORFeomes for some model organisms have been constructed following the completion of their genome sequencing projects. These libraries are based on Gateway cloning technology, which facilitates the study of protein function by simplifying the subcloning of open reading frames (ORF) into any suitable destination vector. The expression of proteins of interest as fusions with functional modules is a frequent approach in their initial functional characterization. A limited number of Gateway destination expression vectors allow the construction of fusion proteins from ORFeome-derived sequences, but they are restricted to the possibilities offered by their inbuilt functional modules and their pre-defined model organism-specificity. Thus, the availability of cloning systems that overcome these limitations would be highly advantageous.ResultsWe present a versatile cloning toolkit for constructing fully-customizable three-part fusion proteins based on the MultiSite Gateway cloning system. The fusion protein components are encoded in the three plasmids integral to the kit. These can recombine with any purposely-engineered destination vector that uses a heterologous promoter external to the Gateway cassette, leading to the in-frame cloning of an ORF of interest flanked by two functional modules. In contrast to previous systems, a third part becomes available for peptide-encoding as it no longer needs to contain a promoter, resulting in an increased number of possible fusion combinations. We have constructed the kit’s component plasmids and demonstrate its functionality by providing proof-of-principle data on the expression of prototype fluorescent fusions in transiently-transfected cells.ConclusionsWe have developed a toolkit for creating fusion proteins with customized N- and C-term modules from Gateway entry clones encoding ORFs of interest. Importantly, our method allows entry clones obtained from ORFeome collections to be used without prior modifications. Using this technology, any existing Gateway destination expression vector with its model-specific properties could be easily adapted for expressing fusion proteins.

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In vivo imaging of induction of heat-shock protein-70 gene expression with fluorescence reflectance imaging and intravital confocal microscopy following brain ischaemia in reporter mice

Abstract: This study showed in vivo induction of Hsp-70 gene expression in ischaemic brain using reporter mice. The fluorescence signal showed in vivo the induction of Hsp-70 in penumbra neurons and in the vasculature within the ischaemic core.

Cited by 15 publications

References 43 publications

Alarmin S100A8/S100A9 as a biomarker for molecular imaging of local inflammatory activity

Alarmin S100A8/S100A9 as a biomarker for molecular imaging of local inflammatory activity

Neuronal PirB Upregulated in Cerebral Ischemia Acts as an Attractive Theranostic Target for Ischemic Stroke

A plasmid toolkit for cloning chimeric cDNAs encoding customized fusion proteins into any Gateway destination expression vector

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