Stem cell models of disease
Pluripotent stem cell derived neurons expressing green fluorescent protein (under the control of LMX1A promoters) and counter labelled with antibodies to a non-specific neuronal marker (β3 tubulin, in white) and a marker of nuclei (blue). These cells were isolated using green fluorescence activated cell sort 15 days before immunolabeling. While many remain green, it is clear that some are not. We are currently investigating how neurons respond to their extracellular environment in health and disease.
Stem cells can self-renew indefinitely and, being pluripotent, they can develop into any cell type present in the adult. Much of the focus of stem cell biology is directed at the specification of particular cellular phenotypes (neurons, cardiomyocytes, blood, etc.) for use in cell replacement therapies, drug screening and disease modelling.
My research is largely focussed on the development of models of neuropsychiatric and neurodegenerative diseases. The first step in this model development program is defining conditions that permit the generation of specific cell types (for example, A9 or A10 midbrain dopaminergic neurons), the second step is defining conditions that reflect the environment associated with disease. This simple approach enables us to track the progression of healthy cells to unhealthy states. Understanding disease pathology enables us to efficiently direct studies toward new therapeutic targets.
The Stem Cell Group at MIPS has generated a number of embryonic stem cell reporter lines enabling the identification of particular neurons in culture. The development of these reporter lines underlies our core interest in developing human stem cell based models of human disease; however, these fluorescent reporter lines have a broader significance and we have used them to facilitate (i) the identification of conditions conducive to the enrichment of specific neural phenotypes, (ii) the characterization of neurons in culture and (iii) the development of models of neurodegeneration and neuropsychiatric disease.
Haynes, J.M., & Halliwell, R.F. (2017) Editorial: The Potential of Stem Cells for 21st Century Neuroscience II. Neurochem Int. 2017 Jun;106:1-2. doi: 10.1016/j.neuint.2017.05.001.
Hunt, C.P.J., Pouton, C.W. & Haynes, J.M. (2016) Differentiation and characterization of human DARPP32 cortical neurons from embryonic stem cells. Neurochem Int. S0197-0186(17)30003-7. doi: 10.1016/j.neuint.2017.01.003.
Niclis, J., Gantner, C., Alsanie, W., McDougall, S., Bye, C., Elefanty, A., Stanley, E., Haynes, J.M., Pouton, C.W.P. Thompson, L., Parish, C. (2017). Efficiently specified ventral midbrain dopamine neurons from human pluripotent stem cells uder xeno-free conditions restore motor deficits in Parkinsonian rodents. Stem Cells Translational Medicine 6:937-948. doi: 10.5966/sctm.2016-0073.
Lagerqvist, E.L., Finnin, B.A, Elliott, D.A., Anderson, D.J., Wu, S., Pouton, C.W. & Haynes, J.M. (2015) Comparing mouse and human pluripotent stem cell derived cardiac cells: both systems have advantages for pharmacological and toxicological screening. J Pharm Meth Tox. 74:17-25.
Watmuff, B., Hartley, B.J., Hunt, C.P.J, Fabb, S.A., Pouton, C.W. & Haynes, J.M. (2015). Human pluripotent stem cell derived midbrain PITX3eGFP/w neurons: a versatile tool for pharmacological screening and neurodegenerative modelling. Frontiers in Cellular Neuroscience. doi: 10.3389/fncel.2015.00104.
Chan, L.J., Bulitta, J.B., Williams, C.C., Ascher, D., Haynes, J.M., McLeod, V.M., Porter, C.J.H., Kaminskas, L.M. (2015) PEGylation does not significantly change the initial intravenous or subcutaneous pharmacokinetics or lymphatic exposure of trastuzumab in rats, but increases plasma clearance after subcutaneous administration. Molecular Pharmaceutics 12(3):794-809.
Kaminskas, L., McLeod, V., Ascher, D., Ryan, G., Jones, S., Haynes, J.M., Trevaskis, N., Chan, L., Sloan, E., Finnin, B., Williamson, M., Velkov, T., Williams, E., Kelly, B., Owen, D., Porter, C. (2015) Methotrexate-conjugated PEGylated dendrimers show differential patterns of deposition and activity in tumour-burdened lymph nodes after intravenous and subcutaneous administration in rats. Molecular Pharmaceutics 12(2):432-43. doi: 10.1021/mp500531e.
Tsang, K.M.C., Annabi, N., Zhou, K., Ercole, F., Karst, D. Haynes, J.M., Evans, R.A., Thissen, H., Khademhosseini, A. & Forsythe, J.S. (2014) Facile One-step Micropatterning Using Photodegradable Gelatin Methacrylate Hydrogels for Improved Cardiomyocyte Organization and Alignment. Advanced Functional Materials. 25(6):977–986.
Skelton, R.J.P., Costa M., Anderson D.J. Bruveris F., Finnin B.A., Koutsis K., Arasaratnam D., White A.J., Rafii A., Ng E.S., Elefanty A.G., Stanley E.G., Pouton C.W., Haynes J.M., Ardehali R., Davis R.P., Mummery C.L. & Elliott D.A. (2014). SIRPA, VCAM1 and CD34 identify discrete lineages during early human cardiovascular development. Stem Cell Res. 13(1):172-9.
Kaminskas, L. McLeod, V.M. Ryan, G.M., Kelly, B.D., Haynes, J.M., Williamson, M. Thienthong, N., Owen, D.J., Porter, C.J. (2014). Pulmonary administration of a doxorubicin-conjugated dendrimer enhances exposure of lung metastases to drug and improves cancer therapy, Journal of Controlled Release. 183:18-26.
Hartley, B.J., Fabb, S.A., Finnin, B.A.L., Haynes, J.M., Pouton, C.W. (2014). Zinc-finger Nuclease Enhanced Gene Targeting in Human Pluripotent Stem Cells. J Vis Exp. 2014 Aug 23;(90):e51764.
Hunt, C.P.J., Watmuff, B., Hartley, B.J., Pouton C.W. & Haynes, J.M. (2014) Genetic reporter cell lines: tools for stem cell biology and drug discovery. 'Neural Stem Cell Assays' Ed: Kaur, N and Vermuri, M C, John Wiley & Sons.
Hartley, B.J., Watmuff, B., Hunt, C.P.J., Haynes, J.M. & Pouton, C.W. (2014) In vitro differentiation of pluripotent stem cells towards either forebrain GABAergic or midbrain dopaminergic neurons. 'Neural Stem Cell Assays' Ed: Kaur, N and Vermuri, M C, John Wiley & Sons.
Watmuff, B., Hartley, B.J., Pouton, C.W., Hunt C.P.J. & Haynes, J.M. (2013) Using stem cell-derived dopaminergic neurons to model neurodegeneration. Future Neurology. 8(6): 649 -661.
Hunt, C.P.J., Fabb, S.A., Pouton C.W. & Haynes, J.M. (2013) DNA-dependent protein kinase is a context dependent regulator of Lmx1a and midbrain specification. PLoSOne;8(10):e78759.
Oliver, V.L., Poulios, K., Ventura, S. & Haynes, J.M. (2013) A novel androgen signalling pathway uses dihydrotestosterone, but not testosterone, to activate the epidermal growth factor receptor signalling cascade in prostate stromal cells. Br. J. Pharmacol. 170(3): 592-601.
Niclis, J.C., Pinar, A., Haynes, J.M., Alsanie, W., Jenny, R. & Cram D.S. (2013) Characterization of forebrain neurons derived from late-onset Huntington's disease human embryonic stem cell lines. Frontiers in Cellular Neuroscience. 7(37): Epub doi: 10.3389/ fncel.2013.00037.
- Matt Blurton-Jones (USC)
- Jon Niclis, Clare Parish, Lachlan Thompson (Howard Florey Institute)
- Professor Colin Pouton (MIPS)
- Sharon Ricardo (Faculty MNHS, Monash University)
- John Forsythe (Monash Engineering)
- Carmel O'Brien (CSIRO)
- David Elliott (MCRI)
- Dr Katie Leach (Drug Discovery Biology, MIPS)
Dr Joan Ho
Former student lab members
- Ashley Preston (graduated PhD 2003)
- Anna Cook (PhD, 2004)
- Simer Khaira (PhD 2009)
- Christian Nefzger (PhD 2009)
- Angela Xie (MSc 2009)
- Ebba Louise Lagerqvist (PhD 2012)
- Victoria Oliver (PhD 2012)
- Colin Su (PhD 2012)
- Rhys Bellinge (MSc 2013)
- Pankaj Gulati (PhD 2013)
- Brigham Hartley (PhD 2013)
Past staff lab members
- Nathalie Touchon-Danguy
- Warren Raye
- Hady Warden
- 2015-2018 NHMRC Optimizing the differentiation and expansion of microglial progenitors from human pluripotent stem cells for the study and treatment of neurological disease. Pouton & Haynes.
- 2017 Alport Foundation of Australia. Dysregulation of focal adhesion kinase in Alport Syndrome patients. Haynes & Ricardo.
- 2014-2016 ARC Control of cell fate decisions in neurogenesis: use of embryonic stem cells to investigate key signalling systems and gene expression programs. Pouton & Haynes.
- 2014 Using induced pluripotent stem cell-derived podocytes to create an in vitro model of Alport syndrome.
- 2010-2013 California Institute of Regenerative Medicine (CIRM)/Victorian Government Stem Cell Alliance. (DIIRD funding). "Developmental Candidates" for Cell-Based Therapies for Parkinson's Disease (PD).
- 2010-2011 Australian Stem Cell Centre. Determination of the most promising strategy for cell therapy of Alzheimer's disease: significance of cholinergic neurogenesis versus neural 'chaperone' activity to recovery of cognitive function following hippocampal transplantation of ES cell-derived neural stem cells or neural progenitors.
- 2010-2011 Australian Stem Cell Centre. Stem cell derived cardiomyocytes: tools for investigating cardiac disease.
- 2007-2010 Pfizer / Australian Government P3 scheme: Discovery of low molecular weight inducers of neural differentiation in mouse ES cells.
- 2009-2010 Heart Foundation. A vicious cycle involving adipokines, oxidative stress and renal sympathetic neuroeffector dysfunction in obesity.
- 2007-2009 ARC Linkage project. Functional characterisation of neurons derived from embryonic stem cells and NS cells.
- 2005-2006 Heart Foundation. Angiotensin II-induced vasodilatation in the renal medulla: physiological significance and cellular signalling mechanisms.
- 2004-2007 ARC Linkage. Neurons isolated from embryonic stem cells as functional models for drug discovery.