Pharmacology and Clinical Pharmacology

Protein aggregates linked to C9ORF72 gene mutations in motor neuron disease- a human tissue study

Supervisor

Emma Scotter
Kevin Lee

Discipline

Pharmacology and Clinical Pharmacology

Project code: MHS011

Background
Motor neuron disease is a fatal and incurable movement disorder affecting ~1 in 15,000 New Zealanders. The most common genetic cause of motor neuron disease, accounting for around 10% of cases, is a nucleotide repeat mutation in the gene C9ORF72. We previously characterised three types of protein aggregates in motor neuron disease cases caused by C9ORF72 gene mutations (Scotter et al., 2017). This project seeks to determine whether in these C9ORF72 mutant cases, brain cells containing one type of aggregate are predisposed to accumulating another type of aggregate, and explore the temporal sequence of protein deposition. The project will test a method combining immunofluorescence staining with single cell labelling by microelectrode dye injection. We will also compare the morphology of filled cells with or without aggregates. By determining which protein is nucleating or seeding protein aggregation, we hope to determine an upstream therapeutic target for reducing the protein aggregation ‘domino effect’.

Skills
Human brain tissue immunofluorescence
-Human brain tissue microinjection
-Semi-automated imaging
-Automated image analysis
-Software: Metamorph, Microsoft Excel, statistical and graphing software
-Scientific writing

The student will be based at the Centre for Brain Research, University of Auckland. This pilot project is part of a larger program of research and there is potential for a successful candidate to continue into a Masters or PhD project.

Do blood sera from people with motor neuron disease harbour blood-brain barrier leakage factors or neurovascular toxins?

Supervisor

Emma Scotter
Mike Dragunow

Discipline

Pharmacology and Clinical Pharmacology

Project code: MHS019

Background
Motor neuron disease is a fatal and incurable movement disorder affecting ~1 in 15,000 New Zealanders. We have collected blood sera from a pilot cohort of controls and people living with motor neuron disease. This project seeks to test whether these blood sera show evidence of neuroinflammatory processes or blood-brain barrier leakage. Sera will also be applied to live human brain pericytes in order to assess whether the degeneration of pericytes in MND relates to their exposure to serum with toxic properties. This work complements a wider program of work investigating blood-brain and blood-spinal cord barrier leakage in motor neuron disease using a range of human tissues.
Skills
- Proteome profiler arrays
- Mammalian cell culture
- Immunofluorescence
- Semi-automated imaging and analysis
- Software: Metamorph, Microsoft Excel, statistical and graphing software
- Scientific writing

The student will be based at the Centre for Brain Research, University of Auckland.
This pilot project is part of a larger program of research and there is potential for a successful candidate to continue into a Masters or PhD project.

Functional Consequences of a Disease-Implicated Single Nucleotide Polymorphism

Supervisor

Natasha Grimsey
Caitlin Oyagawa

Discipline

Pharmacology and Clinical Pharmacology/Centre for Brain Research

Project code: MHS083

Cannabinoid Receptor 2 (CB2), so named because it is one of the receptors that responds to the active components in Cannabis sativa, is a G Protein-Coupled Receptor with considerable, but as yet untapped, therapeutic potential.

Expressed primarily in the immune system, CB2-targeted drugs are promising therapeutic leads in a wide range of disorders involving immune system dysregulation, including multiple sclerosis, autoimmune disorders, atherosclerosis, stroke and inflammatory bowel disease. Meanwhile, the likely impact of inflammation in the pathogenesis of mental illness is gaining recognition and a neuroimmunological and inflammatory basis has been proposed for a wide range of psychiatric disorders. Given CB2’s ability to modulate the immune system it seems an attractive candidate target in these disorders – or indeed, may exhibit dysfunction in these conditions which may contribute to pathogenesis.

A naturally occurring non-synonymous single nucleotide polymorphism (SNP) in CB2 has been reported to have increased incidence in bipolar disorder. Our pilot experiments on this polymorphism suggest it exhibits unique molecular pharmacology in comparison with “wild-type” CB2. This project will investigate this observation further and probe the functional consequences of this receptor modification.<br><br>

Skills Taught/Utilised<br>
• Mammalian cell culture and transfection<br>
• GPCR signalling assays, e.g. cAMP, beta-arrestin<br>
• Immunocytochemistry<br>
• Data analysis, graphing, statistics; scientific writing<br>

A student with background and interest in molecular pharmacology (eg. MEDSCI204, 304) is preferred, but this would also be suitable for students with general interests in mammalian cell biology and/or immune- or neuroscience-related therapeutic development/application (eg. BIOSCI201, 203, 353, 349, MEDSCI202, 307, 314).

Honours, Masters and PhD projects are also available in 2019 in related areas of study.

Investigating New Drugs for a Receptor with Untapped Therapeutic Potential

Supervisor

Natasha Grimsey
Yurii Saroz

Discipline

Pharmacology and Clinical Pharmacology/Centre for Brain Research

Project code: MHS084

G protein-coupled receptors (GPCRs) are the targets of more than one third of currently approved pharmaceuticals. The need to enhance and unlock new therapeutic effects, but reduce unwanted side-effects, is driving research to improve drug specificity and fine-tune drug action. While traditional drug discovery and design are focused on simply activating/inactivating receptors, we now know that each GPCR is pleiotropic in nature and exhibits considerable complexity in its potential for modulating cell function. This complexity represents untapped potential in drug development and lays in wait to be harnessed for therapeutic gain.

Cannabinoid Receptor 2 (CB2) is a GPCR with considerable, though as yet unrealised, therapeutic potential. Predominantly expressed in the immune system, CB2 activation is widely associated with immune suppression, and is therefore of particular interest as a therapeutic target in a diverse range of disorders involving immune system over-activation, including auto-immune diseases and neuroinflammation, as well as in various cancers, cardiovascular disease, stroke, mental illness, and osteoporosis.

We are characterising novel CB2 ligands synthesised by medicinal chemists with whom we are collaborating. Our pilot data suggests that these ligands activate unique signalling patterns via CB2 and therefore may represent interesting therapeutic leads. This project will extend research on these novel compounds to as yet untested signalling pathways, including arrestin recruitment, potassium channel activation and desensitisation, and calcium flux (range of assays utilised will be time-dependent).

Skills Taught/Utilised
• Mammalian cell culture and transfection<br>
• GPCR signalling assays, e.g. beta-arrestin, potassium channel, intracellular calcium
• Data analysis, graphing, statistics; scientific writing

A student with background and interest in molecular pharmacology (eg. MEDSCI204, 304) is preferred, but this would also be suitable for students with general interests in mammalian cell biology, medicinal chemistry / drug development and/or immune-related therapeutic application (eg. BIOSCI201, 203, 353, 349, CHEM390, 392, MEDSCI202, 314). Honours, Masters and PhD projects are also available in 2019 in related areas of study.

Slipping away – Limiting tumour cell migration in malignant brain tumours

Supervisor

Dr Thomas Park
Professor Mike Dragunow

Discipline

Pharmacology and Clinical Pharmacology

Project code: MHS087

Glioblastoma multiforme (GBM) is the most common and fatal brain tumour with current clinical practice consisting of surgical resection, followed by chemo- and radiation therapy. Despite these therapies, the prognosis is poor due to the aggressive invasion of the surrounding brain tissue by highly migratory treatment-resistant GBM cells. This study seeks to study the underlying mechanisms governing cellular migration of these treatment-resistant GBM cells. Patient-derived GBM cells will be studied to elucidate these mechanisms with a focus on the pro-migratory molecules that makes these cells ‘slippery’.

The student will be working at the Centre for Brain Research with patient-derived GBM cells isolated from neurosurgical specimens and will learn cell culture, molecular and imaging techniques.

Investigating brain cell type-specific transcriptomic changes in neurological diseases

Supervisor

Dr Thomas Park
Professor Mike Dragunow

Discipline

Pharmacology and Clinical Pharmacology

Project code: MHS088

The underlying mechanism for many neurological diseases are multifaceted and involve many, if not all the cell types present in the human brain. Classical transcriptomic studies relied on whole brain tissue extracts to identify changes in neurological diseases, largely neglecting the fact that the brain is a multicellular structure. In this study, we will utilise a relatively new technique of isolating cell-type specific nuclei from frozen human brain tissue to interrogate cell-specific changes that might occur in neurological diseases.

The student will be working with donated adult human brain specimens that have been cryopreserved in the Centre for Brain Research from various neurological diseases to optimise this nuclei-isolation technique and sort individual cell populations for transcriptomic studies.

This project will involve learning immunohistological techniques, nuclei-isolation from frozen human brain tissues, fluorescence-activated cell sorting (FACS) and quantitative PCR techniques to validate and identify genes of interest.

Platelet-derived growth factor receptorß (PDGFRB)-signalling in human brain cells

Supervisor

Professor Mike Dragunow
Deidre Jansson

Discipline

Pharmacology and Clinical Pharmacology

Project code: MHS201

PDGFRB is a critical receptor for survival/proliferation/migration signalling in perivascular pericytes and also in glioma cells in the brain. In this project, we will use pharmacological, cellular and molecular techniques to dissect out the components of these signalling pathways in cultured primary human brain pericytes and in glioma cells to determine whether signalling is similar or altered in the cell types. These cells will be derived from consenting donors undergoing neurosurgical interventions.

This work will have important implications for understanding factors controlling the proliferation and survival of these cell types and in particular for developing treatments to control the growth and spread of human brain tumours.