SPINAL CORD INJURY RESEARCH UNIT
at the Centre for Brain Research
Established by the CatWalk Spinal Cord Injury Trust - September 2011

Overview
The major role of the Spinal Cord Injury Research Unit is to establish expertise and maintain spinal injury models that can be accessed by researchers from throughout New Zealand working on spinal cord injury and repair. The unit will help to further develop, grow and maintain an ongoing research programme with existing international collaborations and foster new initiatives both in New Zealand and abroad. Adminsitrative support and a research environment for student training will be provided by the Centre for Brain Research’s Integrative Neuroscience Facility. The unit will play a key role in educating bright, young students in spinal injury research and clinical awareness and practice, who will go on to postgraduate research and future clinical positions.

Directors
Professor Louise Nicholson is the principal investigator of the Molecular Neuroanatomy Laboratory in the Centre for Brain Research. She is internationally recognized as an authority on molecular neuroanatomy of the human brain, and has expertise in both cellular neuroanatomy and the molecular measures of neuropathology in both animal models and in human neurodegenerative diseases. Her research group currently focuses on the role of gap junctions in central nervous system injury, repair and neurodegeneration. Professor Nicholson is currently the New Zealand representative on both the Australian Neuroscience Society Council, and the Australia New Zealand Spinal Cord Injury Research Network. She has been a Council member and Secretary of the International Basal Ganglia Society. Professor Nicholson is currently the Associate Dean Research for the Faculty of Medical and Health Sciences.

Dr Simon O’Carroll is a research fellow in the Centre for Brain Research. He has expertise in the use of animal models for spinal cord injury and will lead the running of the unit. Dr O’Carroll’s research interests focus on the role of connexins in spinal cord injury, neuroinflammation, neurodegenerative disease and neurogenesis.

Professor Colin Green has published widely in the fields of gap junction biology and their roles in development, health and disease and has an established research interest in central nervous system and spinal cord injury. Professor Green has a strong focus on translational research and currently holds the W & B Hadden Chair of Ophthalmology and Translational Vision Research and was a founding scientist of CoDaTherapeutics (NZ) Ltd and CoDa Therapeutics, Inc (USA), established to apply the major breakthroughs he and collaborators have made in the role of gap junctions in tissue injury and repair.

Location
The Spinal Cord Injury Research Unit (SCIRU) is located in the hub of the Centre for Brain Research – a purpose built dedicated neuroscience research laboratory, situated in the Grafton Campus of the Faculty of Medical and Health Sciences at The University of Auckland. This laboratory environment is a specialist neuroscience research area, with world-class studies underway investigating human brain disease, stem cells, gene therapy, and drug development. As well as the physical proximity to such ground-breaking studies, SCIRU also links to the wider neuroscience research underway in Auckland University, through the Integrative Neuroscience Facility in the Centre for Brain Research. This collaborative approach means that pre-clinical investigations carried out in the laboratory are underpinned by molecular, cellular and clinical work underway in the rest of the centre.

The Centre for Brain Research is directed by Professor Richard Faull. Richard has an international reputation for his research studies on the normal and diseased human brain and has been awarded New Zealand's highest scientific award, the Rutherford Medal. He is the Patron of the Alzheimer's Foundation (Auckland), Alzheimers New Zealand Charitable Trust and the Huntington's Disease Association (Auckland and Northland), and the Medical Patron of the Motor Neurone Disease Association of New Zealand.

Ethics
All research carried out in the SCIRU must first be approved by the University of Auckland Animal Ethics Committee. This stringent ethics committee ensures that all research carried out in the University adheres to international ethical and safety standards. A panel of experts from across the University assess each research project and guarantee only the highest quality will pass to meet its high standards.
The use of human tissue bequeathed to the Neurological Foundation of New Zealand Human Brain Bank must follow strict ethical guidelines. All research projects must first be approved by the University of Auckland Human Ethics Committee. The team have also developed protocols for sensitive use of the tissue according to tikanga Māori.

Laboratory Safety
SCIRU will be located in a PC1 laboratory, and in a dedicated high containment facility within the VJU Research Unit which means that all practices adhere to rigorous health and safety standards. PC1 ensures that all biological material is handled safely and carefully, so that human health is protected. The high containment facility will allow the use stem cell and gene therapy technologies. All employees and students in the laboratory must undergo training in the safety standards before any research is commenced.

Collaborations
SCIRU is part of the Integrative Neuroscience Facility in the Centre for Brain Research, with the aim to produce research that is cutting-edge and maximises resources. Ideas, techniques and skills are shared within the unit, so that ground-breaking developments in the field of spinal cord injury will be able to influence those in stroke treatment, and vice versa. Our researchers also have numerous national and international collaborations, ensuring that SCIRU is at the forefront of global research. SCIRU welcomes new research collaborations and ideas, with the aim of maximising spinal cord research in New Zealand. Our ultimate hope is finding a cure for spinal cord injury.
 

RESEARCH PROJECT UPDATE
Regulation of cell to cell communication for spinal cord injury - Taking the next step

Overview - September 2011
The team of Dr Simon O’Carroll, Professor Colin Green and Professor Louise Nicholson at the University of Auckland discovered that one of the critical changes that takes place after spinal cord injury is an increase in the number of communicating channels between nerve cells. These channels play a major role in spreading the damage from the site of injury to areas that would otherwise not be affected. Using small molecules called mimetic peptides, developed by the team in their laboratory, they can alter cellular function to close these channels. These tests have shown that they can reduce swelling, inflammation and scar formation and promote nerve cell survival. They are currently testing one of these peptides in an animal model of spinal cord injury. The team has shown that delivery of this peptide prevents inflammation, protects nerve cells from death and leads to improved locomotion. They are currently developing this work further to enable delivery of the peptide via the blood stream. The optimal time point for the delivery of the peptide (window of opportunity and optimal treatment period) and whether modifying the peptide may further improve its efficacy are currently being investigated.

In other related work, they have been using small DNA molecules called antisense to prevent the channel proteins from being made. In collaboration with Professor Jack Kessler from Northwestern University in Chicago they have used nanofibre technology, in conjunction with the antisense, to improve nerve fibre regrowth after spinal cord injury in an animal model. Results have shown extensive regrowth of nerve fibre tracts across the damaged area, even in completely split spinal cords.
Future studies will include the use of human spinal cord tissue. This tissue is available from the Neurological Foundation of New Zealand Human Brain Bank based in the Centre for Brain Research. Work on human tissue will allow the team to determine new targets for intervention in spinal cord injury, as well as confirming findings from animal studies. The potential exists for collaborations with other members of the Centre to use stem cell and gene therapy technologies. Collaborations will also be sought with researchers throughout New Zealand in order to gain a better understanding of the causes of disability following injury and to develop new treatments.

Introduction

We are able to stop the opening of pathological channels between nerve cells, which is one of the critical changes occurring early after injury and which leads to the spread of damage between cells in the spinal cord. We have two ways of doing this. 1) the antisense “gel”, which is a topical application and will be useful for repair strategies and 2) a mimetic peptide, which is able to be delivered via the blood stream and can be used for early intervention after spinal cord injury to reduce damage spread.

Reduce inflammation

In this figure we have two segments of spinal cord removed from a rat and grown in a dish. The top segment has not been treated and the spinal cord tissue has swelled out of the ends of the cord (as marked by the dotted lines) showing there has been inflammation. The bottom piece was treated with the antisense “gel” and inflammation has been prevented as no swelling is seen.

These are images of tissue taken from a rat that received a spinal cord contusion injury and 1 hour later it was treated with the peptide (Left hand image) or with saline (Right hand image). The tissue was stained for the presence of immune cells, which are indicated by the yellow or green colour. As you can see treatment with the peptide almost completely prevents inflammation in the damaged cord.

Reduce scarring
These are more images from contused spinal cords as described above. This time they are stained red for cells called astrocytes, which are involved in forming the scar that occurs after injury. The left hand image was treated with saline and shows very strong red staining around the edge of the damaged area (the black area in the middle). The right hand image was treated with peptide and shows very little red staining around the damaged area. This shows the peptide has reduced scarring, which is important for recovery after injury.

Better outcomes
3 weeks after injury; Control animals had some hind limb movement but it was not weight bearing and not coordinated between left and right while peptide treated animals had some weight bearing movement of their hind limbs that was coordinated between left and right.

Repair
Results of an experiment where peripheral nerve grafting was used to promote nerve cell growth into a damaged cord after injury – a potential repair strategy for spinal cord injury; peripheral nerves were grafted into segments of spinal cord in the presence or absence of the antisense gel. The antisense gel greatly improved the number of nerve cell fibres that grew into the peripheral nerve compared with controls. The growth of nerve fibres is important to allow reconnection of a damaged cord and this shows that the antisense gel may help improve the success of spinal cord repair strategies, by reducing the inflammation and tissue dieback that occurs.

This image shows a piece of damaged spinal cord (left side of the image) into which has been grafted a biologically active nanofibre matrix, which are being used for spinal cord injury research, in the presence of the antisense gel. The area highlighted by the red oval shows the extensive growth of nerve fibres from the spinal cord in to the nanofibre matrix. This growth is much greater than what is seen when no antisense gel is used. This again shows the potential of the antisense gel in enhancing repair strategies for spinal cord injury.