Vanessa Jane Hall
Dyrlægevej 100, 1870 Frederiksberg C, 1-05
My PhD resulted in the production of the very first cloned cows in Australia. I was internationally recognized then for developing an enhanced somatic cell nuclear transfer technique and spotlighted in the media in 2005. In my first postdoc in England, I was producing the world’s first cloned human embryos for the purpose of human embryonic stem cell (ESC) lines, which received considerable media and scientific attention. I have also significantly advanced the field of pluripotent cell signalling in the pig with several internationally recognized articles. My interest in applying ESC research for investigating disease began in Sweden and transferred from Parkinson’s disease to Alzheimer’s disease (AD), upon moving to Denmark, where I have now established a unique in-vitro cell model from a pig model of AD, which recapitulates disease pathology. I am currently acting as PI on two research projects in the field of AD with a focus on improving the quality of neurons and production of more disease-relevant cell types produced from patient-derived induced pluripotent stem cells.
Current Research Interests:
- Characterization of the entorhinal cortex in the fetal porcine brain
- Improving differentiation of human induced pluripotent stem cells into relevant neurons for studying Alzheimer's disease
- Characterization of pluripotency in the porcine stem cell populations of the pre-implantation embryo.
- Establishment and characterization of porcine embryonic stem cells and induced pluripotent stem cells.
- Analysis and differentiation of neural stem cells derived from Alzheimer biomedical pig.
Current funded Research projects 2017:
- Danish Research Council (Technology and Production): 2.591 million dKK
- Innovation Fund Denmark (www.brainstem.dk)
1. Use and application of the developing pig brain for producing human stellate cells
Problem statement: Stellate cells are a predominant neuron in the entorhinal cortex (EC), which are lost very early on in Alzheimer’s disease, but despite this, have been poorly studied. If we could produce this cell type from induced pluripotent stem cells (iPSCs), we end up with a very relevant neuron type affected which could help unfold mechanisms that arise early on in the disease.
Hypothesis: Superior grade stellate cells can be produced from human iPSCs using a state-of-the art in-vitro differentiation tool modelled on embryonic development of the porcine brain.
Impact of findings: The impact of this research will be large for the field of brain development, as we will uncover the proteome of the developing cortex and EC in a large mammal, with huge implications for human cortex development. Our findings will impact research in the Alzheimer’s disease area, as we will generate a highly disease-relevant cell type that has never been produced or studied in detail before. Stellate cells of the EC are also considered the border cell contributing to spatial memory, and therefore will also be of interest for neuroscience research on spatial memory processing and grid cell networks.
2. Analysis of nucleolar function in AD.
Purpose: To study how defects in PSEN1 might affect nucleolar function in neurons and glia.
- Mutations in PSEN1 disrupt ribosomal RNA transcription, processesing and assembly.
- Nucleolar stress is increased in neurons and glia derived from PSEN1 patients.
- Defects in nucelolar function result in activation of the nucelolar stress pathway in PSEN 1 AD patients.
Background: A large cohort of Alzheimer’s disease (AD) patients carrying mutations in the presenilin 1 (PSEN1) gene have the most severe form of the disease with earliest age of onset and earliest age of death. Known clinical symptoms and pathology linked to PSEN1-associated AD includes progressive dementia, parkinsonism, notch signaling defects and accumulation of Amyloid-beta (Ab). PSEN1 forms part of the catalytic core of the g-secretase complex, which is important for cleaving transmembrane proteins at the cell membrane such as amyloid precursor protein (APP) and mutations in PSEN1 lead to disruption of this cleavage process and accumulation of toxic forms of Ab.
It has been shown that nucleolin (NCL) (a protein involved in pre-RNA transcription and ribosome assembly) binds to the 3’ non-translated region of APP and may also be dysregulated as a consequence. NCL is a phosphoprotein involved in the synthesis and maturation of ribosomes and expressed in the fibrillar center of the nucleolus. In neurons, NCL has also been shown to be expressed in the cytoplasm, and what it’s role is in the cytoplasm, is unknown. Nucleolar function is important for cell survival. The nucleolus is also important for controling mitochondrial activity and stress signalling pathways, which are both affected in the disease. Furthermore, phosphorylated NCL is present in neurofibrillary tangles (NFTs) and acts as an early marker for NFT. It is clear that a better understanding of the role of NCL and nucleolar function in AD may help to unravel novel key processes that occur in the disease.
1. Investigate nucleolar function. Nucleolar compartments including the fibrillar center, the dense fibrillar centre and the granular components will be investigated in neuroprogenitors, neurons and glia from PSEN1 AD patients by use of transmission electron microscopy.
2. Expression of ribosomal RNA synthesis and processing will be investigated by evaluation of ribosomal RNA genes, PolI and UBF (important for synthesis of ribosomal RNA genes, 28S, 18S and 5S), PolII (important for synthesis of mRNA) and PolIII (important for transcribing 5S and tRNA genes) in the same cells.
3. Investigate the role of NCL. Analysis of expression of NCL will be performed in neuroprogenitors, neurons and glia from PSEN1 AD patients. Expression levels and localization of protein will be determined.
4. Investigating the nucleolar stress and oxidative stress pathway. Expression of ribosomal genes, MDM2 and p53 will be investigated in neuroprogenitors, neurons and glia from PSEN1 AD patients. Expression of JNK2 and TIF1A will be also be investigated.
Previously funded Research Project 2010-2014:
Generation of induced pluripotent stem cells from the APPsw biomedical pig.
Objective of the Project:
The major objective was to produce porcine neural stem cells from wildtype and APPsw transgenic pigs and differentiate these into neurons. The APPsw pig, was produced by Aarhus University and carries a human mutant gene which leads to a juvenile onset of Alzheimer’s disease in humans. The wildtype iPSC-derived neural cells could be used for future transplantations into the Alzheimer’s pig brain and neural cells derived from Alzheimer biomedical pig will be studied in-vitro to investigate the etiology of the disease.
Porcine induced pluripotent stem cells – Generation of in-vitro models and therapeutic neurons for the Alzheimer’s biomedical pig.
Det Frie Forskningsråd: Teknologi og Produktion
The Danish Research Council for Technology and Production Sciences
PhDs (Acted as official co-supervisor for four students)
2014 – 2015 Carlota Pires/University of Copenhagen, Denmark
2011 – 2015 Dong Li: Submitted September 2015/University of Copenhagen, Denmark
2008 – 2011 Yu Gao/Uni of Copenhagen, Denmark: Now a postdoc at University of Wisconsin-Madison
2007 – 2010 Mikkel Rasmussen/Uni of Copenhagen, Denmark Now a postdoc at Bioneer A/S, Denmark developing iPSCs from patients.
Masters and Bachelors
Acted as co-supervisor for 4 masters students (12-18 month projects) and main supervisor for 3 Bachelor’s projects
Fields of interest
Field: Human neural development: Professor Kjeld Møllgård, University of Copenhagen/Denmark
Field: Human neural stem cells in Parkinson’s disease: Associate Professor Morten Meyer, University of Southern Denmark/Denmark
Field: Human ESC transplantation in Parkinson’s disease: Dr. Agnete Kirkeby, Lund University/ Sweden
Field: Induced pluripotent stem cells: Prof. Heiner Niemann, Institute of Farm Animal Genetics, /Germany
Field: Induced pluripotent stem cells: Prof. Andras Dinnyes, Biotalentum/Hungary