Croatian Science Foundation
CANDiD: Characterisation of aggregating proteins in neuropsychiatric diseases, including Drosophila models”
ISkrEN: Istraživanje shizofrenije kroz ekspresiju netopivih proteina
Project leader: Dr. Nicholas Bradshaw, PhD
Project associates: Prof. Rozi Andretić Waldowski, MD PhD, Prof. Gordana Rubeša, MD PhD, Dr. Aristea Pavešić Radonja, MD, Beti Zaharija, MSc, Bobana Samardžija, MSc, Maja Juković MSc, Simone Ruhije Bertoša MSc, Giovanna Dashi, BSc, Tina Fartek, BSc, Anja Hart, BSc.
Duration: 11/2018-03/2023
Funder: Croatian Science Foundation (Hrvatska zaklada za znanost, HRZZ), project grant IP-2018-01-9424
Duration: 11/2018-03/2023
Funder: Croatian Science Foundation (Hrvatska zaklada za znanost, HRZZ), project grant IP-2018-01-9424
While mental illnesses such as schizophrenia, major depression and bipolar disorder have devastating effects on patients and those around them, we still know relatively little about their biological causes. A major problem has been that the genetic background of these illnesses is extremely complex: with many different genes involved, but few having more than a small individual effect on whether someone develops the illness or not. We, along with our collaboration partners, have therefore been working on an alternative method of studying the biology of these disorders. In many other diseases of the brain, such as Alzheimer’s disease or Parkinson’s disease, specific proteins fold incorrectly in the brain, and instead build up in unfolded clusters in the brain, called aggregates. Some of these aggregates are toxic, and actively kill the cells of the brain, while others instead prevent the cells from functioning normally. We and others have therefore been investigating whether similar protein aggregates appear in the brains of patients with life-long mental illnesses. So far, a total of five proteins have been found to aggregate in schizophrenia and/or other conditions, named CRMP1, DISC1, dysbindin-1, NPAS3 and TRIOBP-1.
Our HRZZ project therefore has five main aims:
1) We know that in the case of TRIOBP-1 and NPAS3, very specific sections of the protein are required for them to aggregate – with deletion or mutation of these sections affecting whether or not they form aggregates. Using molecular and cellular biology techniques, we are now investigating whether similar “aggregation critical” sites exist on the other three proteins, to better understand how aggregation occurs.
2) Each of the proteins that we study has been seen to form aggregates alone in mental illness, but there is also evidence, at least in some cases, that they can also bind to each other to form “co-aggregates” which might jointly affect neurons, and ultimately mental illness. We are therefore systematically investigating the interactions, and co-aggregation, of these proteins.
3) Once we know that aggregation does occur in mental illness, the next stage is to determine whether aggregation itself can directly affect the way we think and our behaviour. As a first test of this, we are collaborating with the laboratory of Dr. Rozi Andretić Waldowski to test the effect of aggregation of our proteins in a simple organism: the fruit fly Drosophila.
4) While it is interesting to know if each of our proteins can affect neurons or fruit flies when they begin to aggregate, our aim is to understand how they co-operate together to have effect on mental illness. We will therefore study the affects of these proteins in combination on cell function and behaviour.
5) The study of protein aggregation in mental illness is still very new, and has been reported so far in only a small number of patients. Therefore, through collaborations with the Psychiatry Clinic of our local hospital, along with research institutes abroad, we are continuing to investigate the presence of protein aggregates in samples donated by patients with a variety of mental illnesses.
So far, the following publications have arisen from this project: 1) Zaharija, Samardžija & Bradshaw 2020, 2) Samardžija, Pavešić Radonja et al 2021, 3) Zaharija, Odorčić et al. 2022, 4) Zaharija & Bradshaw 2023.
More details can be found on the Publications page.
More details can be found on the Publications page.
Additionally, Beti Zaharija and Bobana Samardžija are being supported by a PhD student stipends from the HRZZ: DOK-2018-09-5395 (2019-2023) and DOK-2020-01-8580 (2020-2024)
Alexander von Humboldt Foundation
DISC1: Its Structure, Causes of Aggregation and Relevance to Disease (the DISCARD collaboration)
Project leaders: Dr. Nicholas Bradshaw, PhD and Dr. Oliver Weiergräber, PhD
Project associates: Dr. Abhishek Cukkemane PhD, Beti Zaharija, MSc, Bobana Samardžija, Priyanka Sathyamoorthy MSc
Duration: 02/2022-01/2025
Funder: Alexander von Humboldt Foundation/Stiftung
Chronic mental illnesses, such as schizophrenia and major depressive disorder, are debilitating conditions, whose pathological basis is complex and still poorly understood. One protein of significant interest in studying these is Disrupted in Schizophrenia 1 (DISC1), a scaffolding protein involved in neurodevelopment, which is not only implicated through genetic evidence, but has also been shown to form misfolded protein aggregates in the brains of patients. These aggregates may be analogous to similar events in neurodegenerative conditions, such as Parkinson's disease or ALS.
The three-dimensional structure of the DISC1 protein is poorly understood and has proven very difficult to study. A breakthrough was made recently, however, by using a high throughput approach to map four putative folded domains within the protein.
In this project, a collaboration between researchers at the University of Rijeka (Croatia) and the Forschungszentrum Jülich (Germany), we aim to analyse both the structure of the DISC1 protein and the process by which it forms aggregates, using an interdisciplinary approach. At the University of Rijeka (Croatia), we are using molecular and cellular biology approaches to confirm and refine the domain structure of DISC1 in mammalian cells, as well as study the cellular consequences of its aggregation, through the expression of fragments of DISC1 in mammalian cells. Our partners in Jülich (Germany), meanwhile, are probing the structure and aggregation dynamics of the same fragments using biophysical and structural biology approaches in vitro. This is facilitated using a shared plasmid cloning program, as well as reciprocal research visits to coordinate approaches and share skills, including three extended visits by doctoral students. Both parts of the work are helped through regular online collaboration, and regular exchange visits by members of both laboratories to their partners abroad.
In this way, we aim to understand not only the structure of the DISC1 protein, but also how this structure goes wrong in mental illness, leading to the eventual onset of clinical symptoms.
Project leaders: Dr. Nicholas Bradshaw, PhD and Dr. Oliver Weiergräber, PhD
Project associates: Dr. Abhishek Cukkemane PhD, Beti Zaharija, MSc, Bobana Samardžija, Priyanka Sathyamoorthy MSc
Duration: 02/2022-01/2025
Funder: Alexander von Humboldt Foundation/Stiftung
Chronic mental illnesses, such as schizophrenia and major depressive disorder, are debilitating conditions, whose pathological basis is complex and still poorly understood. One protein of significant interest in studying these is Disrupted in Schizophrenia 1 (DISC1), a scaffolding protein involved in neurodevelopment, which is not only implicated through genetic evidence, but has also been shown to form misfolded protein aggregates in the brains of patients. These aggregates may be analogous to similar events in neurodegenerative conditions, such as Parkinson's disease or ALS.
The three-dimensional structure of the DISC1 protein is poorly understood and has proven very difficult to study. A breakthrough was made recently, however, by using a high throughput approach to map four putative folded domains within the protein.
In this project, a collaboration between researchers at the University of Rijeka (Croatia) and the Forschungszentrum Jülich (Germany), we aim to analyse both the structure of the DISC1 protein and the process by which it forms aggregates, using an interdisciplinary approach. At the University of Rijeka (Croatia), we are using molecular and cellular biology approaches to confirm and refine the domain structure of DISC1 in mammalian cells, as well as study the cellular consequences of its aggregation, through the expression of fragments of DISC1 in mammalian cells. Our partners in Jülich (Germany), meanwhile, are probing the structure and aggregation dynamics of the same fragments using biophysical and structural biology approaches in vitro. This is facilitated using a shared plasmid cloning program, as well as reciprocal research visits to coordinate approaches and share skills, including three extended visits by doctoral students. Both parts of the work are helped through regular online collaboration, and regular exchange visits by members of both laboratories to their partners abroad.
In this way, we aim to understand not only the structure of the DISC1 protein, but also how this structure goes wrong in mental illness, leading to the eventual onset of clinical symptoms.
Travel and collaboration grants
Students in the Bradshaw group have so far been highly successful in winning ERASMUS+ traineeships. Beti Zaharija performed an 8-month traineeship in 2019 to the University of Edinburgh, to work in the group of Dr. Kirsty Millar, while both her and and Bobana Samardžija have performed traineeships at the Jülich Research Centre (Forschungszentrum Jülich) under the supervision of Dr. Oliver Weiergräber. Nick Bradshaw has also undertaken a teaching exchange to the University of Bremen in 2022.
In spring 2023, Nick Bradshaw undertook a CAPES PrInt visiting professorship to the Federal University of São Paulo (UNIFESP), in Brazil, funded by the Brazilian government. This involved teaching two courses, on communication in English and mental illness research, as well as working on collaborative research with Prof Mirian Hayashi.
The Bradshaw group has also recently won a bilateral project from the Ministry of Science and Education (MZO) for travel to support collaborative research with Prof Alja Videtič Paska from the University of Ljubljana. Together, we will investigate protein aggregates as potential biomarkers for suicidal tendency.
Equipment grants
Equipment throughout the Department of Biotechnology, including in our labs was funded by the European Union, through the European Regional Development Fund grant “Razvoj istraživačke infrastrukture na Kampusu Sveučilišta u Rijeci” (Developing research infrastructure at the Campus of the University of Rijeka, RC.2.2.06.0001)
Additionally, as an alumni of the Alexander von Humboldt Foundation (postdoctoral fellowship 2011-2014), Dr. Bradshaw received an equipment subsidy from the Foundation to help establish the research group, and which was presented by the Political Attaché of the German Embassy in Croatia.
Previous funding
Our group has received two grants from the University of Rijeka to support our work (young researcher’s grant 969, stimulation funding 1157). Previous funding to Dr. Bradshaw has come from the Fritz Thyssen Foundation, the Alexander von Humboldt Foundation and the Heinrich Heine University Düsseldorf.
Our group has received two grants from the University of Rijeka to support our work (young researcher’s grant 969, stimulation funding 1157). Previous funding to Dr. Bradshaw has come from the Fritz Thyssen Foundation, the Alexander von Humboldt Foundation and the Heinrich Heine University Düsseldorf.
