MicroRNAs signature in Traumatic Brain Injury (TBI): diagnosis, prognosis and functional studies
Valentina Di Pietro, Antonio Belli
We have discovered that several microRNAs are expressed in blood, saliva and urine of TBI patients within minutes of injury. These are differentially expressed in different TBI severities and outcomes, reflecting differential pathophysiological responses to injury. MicroRNAs are silencing and post-transcriptional regulators of gene expression, and capture the multidimentionality of host-response at tissue and systemic level. They modulate downstream processes in brain and systemic targets, and dynamically continue to do so as the pathology evolves. The discovery of a new regulatory signalling into TBI pathology represents a step-change for molecular TBI research.
SCRUM study: Study of Concussion in Rugby Union testing microRNAs
Valentina Di Pietro, Antonio Belli, Patrick Ohalloran, Kamal Yakoub, Conor Bentley, Callum Watson, Ugo Scarpa, David Davis
This study is carried out in association with the Rugby Football Union (RFU), Premiership Rugby and the Rugby Players' Association. During matches in 2017-18, players with confirmed or suspected concussion will provide saliva and urine samples immediately following the injury and will give follow-up samples as they go through the return-to-play protocol. Samples will be used to test microRNAs as biomarkers of concussion. The study will run alongside the existing Head Injury Assessments (HIA) and if the results support those found in laboratory tests, a new pitch-side test on a device will be developed.
Development of TBI point-of-care diagnostic devices
Valentina Di Pietro, Ugo Scarpa, Antonio Belli, Biovici diagnostics Ltd
Over 1.4 million people attend UK Emergency Department every year with a TBI. Many more suffer concussion during contact sports but are not treated in hospital. Timely diagnosis of TBI is key. We have identified highly sensitive, ultra-early biomarkers of TBI consisting of microRNAs. However, a sensitive biomarker would be no used for roadside or pitch-side diagnosis if samples still needed analysing in a hospital lab. We have therefore partnered with Biovici diagnostic Ltd, a company who is developing a rapid portable and low-cost detection method.
Biomarkers of TBI and neuroimaging
Valentina Di Pietro, Antonio Belli, Aron K Barbey (University of Illinois)
Biomarkers (microRNAs/proteins/metabolites) identified in different bio-fluids of our cohort of concussed athletes will be correlated with the most advanced neuroimaging techniques (MRI, fMRI, DTI, MRE, MRS) and a battery of neurocognitive tests available at the Biomedical Imaging Center at the Beckman Institute of Illinois. The integration of biomarkers, neuroimaging and neuropsychology in a single sample would allow for comparison across multiple measurements resulting in refinement of their individual diagnostic, prognostic and clinical utility as well as in evaluating their effectiveness to detect neural and cognitive recovery.
Valentina Di Pietro, Joachim T Operskalski (University of Illinois), Aron K Barbey (University of Illinois)
Saliva samples of TBI patients were collected at the Beckaman Institute together with neuroimaging (MRE, MRI) and neurocognitive assessments. Samples will be used for the analysis of DNA polymorphism and correlated with cognitive and behavioural indicators in relation to neuroimaging. The final aim is to characterize the processes of recovery.
A characterisation of exosomes within different bio-fluids following concussion and potential therapeutic applications
Valentina Di Pietro, Callum Watson, Antonio Belli
Exosomes (40nm-100nm in diameter) are forms of extracellular vesicles (EVs) that affect disease processes through their involvement in inter cell communication. Their composition is varied, containing numerous signalling molecules including DNA, RNA and proteins appropriate for signalling at the target location which if changed could detail important components in concussion phenotype, demonstrating their potential as a clinical biomarker. Exosomes have potential as biomarkers if changes during a disease process are consistent and measureable with two potential methods of monitoring changes; through change in exosome number or change in composition of exosomes. We aim to demonstrate that: concussed patients will have an increase in exosome number which could be objectively measured in different bio-fluids and miRNA profiles will be distinct between concussed and healthy patients and based on the bio-fluid analysed.
MiRNA functional study
Valentina Di Pietro, Adam Thompson, Antonio Belli
Multiple methodologies are now available to ascertain miRNA targeting. The luciferase test is the most used to experimentally validate the bioinformatically predicted mRNA targets. Understanding the biological role of the microRNA differentially expressed in pathological state (such as mild TBI) is quiet relevant to explore and understand the possibility of using miRNA as therapeutic agents or identifying new therapeutic pathways. In this study we want to investigate the biological role of microRNAs differentially expressed in TBI with the aim to use it in the near future as potential therapeutic drug or to identify new area for therapeutic interventions. Understand the biology hidden by this microRNA is fundamental to restore its normal biological function.
Neuro-inflammatory proteins in TBI
Valentina Di Pietro, Antonio Belli
Within minutes of a traumatic impact, a robust inflammatory response is elicited in the injured brain. Neuroinflammation is responsible for both beneficial and detrimental effects, contributing to secondary brain damage but also facilitating neurorepair. For all these reasons, these inflammatory molecules could satisfy the main characteristics required for a biomarker in the diagnosis and prognosis of TBI. In particular, the early identification of these biomarkers could permit clinicians to recognise and treat those patients at risk of secondary injury while they are still capable of responding to therapy before irreversible damages occur.
Changes of metabolism in severe and mild TBI in an in vivo animal model
Valentina Di Pietro, Antonio Belli, Giuseppe Lazzarino (University of Catania), Barbara Tavazzi (Catholic University of Rome)
Using different approaches (HPLC to analyse biochemical alterations of energy metabolism and oxidative/nitrosative stresses, microarray to analyse gene expression profile) the metabolic, enzymatic and gene changes following graded diffuse traumatic brain injury (TBI) are studied in a weight drop model of TBI in rats.
Neurobioreactors: a new in vitro stretch injury device
Valentina Di Pietro, Adam Thomposn, Lisa J Hill, Antonio Belli, Alastair Campbell Ritchie (University of Nottingham).
This project developed a reliable and reproducible platform for testing drugs and other neuroprotective strategies using an in vitro model of organotypic hippocampal slice cultures. The cultures are stretched on a silicone membrane to mimic in vivo tissue deformation experienced during TBI. The novelty of this study is to understand the molecular changes occurring in TBI in a “clean” environment without the systemic confounders of the in vivo model. A prototype of the model has already been built thanks to the ongoing collaboration with Dr Alastair Campbell Ritchie (UoN). The prototype uses pneumatic pressure as the actuation method, giving greater flexibility in stimulation regimens than current methods using direct physical displacement.
Micro-optofluidic engineered device technology (MEDtech) for timely assessment of TBI
Pola Goldberg Oppenheimer, Valentina Di Pietro, Antonio Belli
In this project a highly-innovative, reliable, diagnostic technology for rapid detection of biomarkers at a picomolar range capable of discriminating between different analytes from complex bio-fluids has been developed. The portable, minimally-invasive MEDTech is based on cost-effective, controllable electrohydrodynamic (EHD) sub-microstructures engineered for multiplexed-surface enhanced Raman scattering (SERS) detection. It combines sensitive, reproducible and molecule-specific EHD-SERS lab-on-a-chip with miniaturised optofluidic platforms for on-the-spot biomarkers detection, either individually or as a panel, to enable real-time diagnostics and care-delivery.