CIPL

New Tools for Real-Time Cellular Imaging and Protein Labelling

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 Obiettivo del progetto (Objective)

'The primary aim of the proposed research is to develop smart new imaging tools and reagents for real-time in situ monitoring of specific cellular processes both in vitro and in vivo, with a particular focus on lung function and apoptosis/inflammation. The methods and tools currently available to define the inflammatory/infective state in lung tissues are limited and crude. Therefore, I will design, synthesise and validate novel clinical molecular imaging probes which can, in the future, be used to define the inflammatory and infective aetiology of lung infiltrates in intensive care unit patients. These dual probes would allow clinicians in intensive care units to rapidly and accurately determine the aetiology of deteriorating respiratory function or clinical status of the patient. These activity-based probes are designed to detect caspase, human neutraphil elastase as well as bacterial presence and activity in the target tissue. The research will be conducted at the interface of chemistry, biology and pre-clinical medicine, with molecules being made and then directly applied in pertinent in vivo models. In addition to the imaging work, I will develop an efficient, selective and generic labelling method for proteins.'

Introduzione (Teaser)

Diagnosis of lung inflammation in hospitalised patients is often delayed with fatal consequences. European scientists proposed to develop imaging probes for early detection of lung inflammation and infection to allow clinicians to accurately stratify patients for appropriate therapy.

Descrizione progetto (Article)

Acute lung injury (ALI) is a clinical syndrome mainly encountered in patients hospitalised in the intensive care unit (ICU).

ALI is characterised by heavy neutrophil infiltration in the lungs and often leads to ventilator-associated pneumonia (VAP), a cause of high mortality.

Tracking ALI remains a major diagnostic challenge for critical care clinicians.Seeking to address this issue, the EU-funded ?New tools for real-time cellular imaging and protein labelling? (CIPL) project proposed to develop specialised molecular imaging probes for detecting ALI and VAP.

These activity-based probes were designed to either target neutrophils, the primary cause of ALI inflammation, or gram-negative and gram-positive bacteria.CIPL members tested a labelling moiety targeted against neutrophil enzymes such as caspase and human neutrophil elastase or bacterial equivalents.

In vitro screening tests verified their discriminatory capacity against healthy lung tissue.Additionally, CIPL researchers were interested in developing protein labelling methodology, which could be combined with their imaging probes.

To this end, they utilised the well-established histidine tag as well as modified specific tyrosine residues on the protein.

Detection of the modified proteins was performed by specific probes containing ligands against these tags and a fluorophore for imaging purposes.Although at their infancy, the CIPL optical molecular probes are expected to help identify critical inflammatory events in the lungs of ICU patients promptly.

In the long-term, this method could be applied for rapidly and accurately determining the aetiology of deteriorating respiratory function or the clinical status of patients.