- Department / Institute
- Faculty of Biology
- Subject area
- Neuroscience
- Project title
- Modelling Human Neurodevelopmental Disease with Cerebral Organoids
- Name of supervisor
- Prof. Dr. David Keays
- Number of open positions
- 1
- Language requirements
- Proficiency in English
- Academic requirements
- 4-year Bachelor's plus Master's Degree; at the time of application, the last final exam should have taken place in the past 4 years.
- Study model
- Full doctoral study model: 36 or 48 months
Project description
The human brain is arguably the most complex structure in biology. It’s construction is dependent on a complex cascade of cellular events that include mitotic division, relocation of migrating neurons, and the extension of dendrites and axons. These processes are reliant on a dynamic and functionally diverse microtubule cytoskeleton.
Microtubules form the mitotic spindle enabling the separation of sister chromatids, they facilitate translocation of the nucleus and extension of the leading process during neuronal migration, and microtubule polymers extend and maintain large and longstanding axons in mature neurons. Reflecting their importance mutations in genes encoding for tubulin subunits and microtubule associated proteins cause severe neurodevelopmental disorders. For instance, variants in TUBA1A are known to cause lissencephaly and cerebral palsy, mutations in TUBB2A cause cortical malformations, and substitutions in MAST1 cause microcephaly, autism and corpus callosum phenotypes.
To study the underlying molecular and cellular mechanisms of these diseases the Keays laboratory is exploiting iPSCs and advanced 2D and 3D neuronal cultures. This project will utilise our recently created biobank of patient derived iPSCs (www.tubulinbiobank.org), coupled with CRISPR-cas9 genome engineering to generate isogenic controls. This project with focus on TUBB2A which is known to cause abnormal cortical gyration, microcephaly, and/or autism. Following the generation of cerebral organoids the student will study how disease causing mutations influence the properties of the microtubule cytoskeleton, and the cellular events necessary for brain formation.
Hypothesis
1) Mutations in TUBB2A act by altering the assembly, stability and/or dynamics of microtubules.
2) Microtubule dysfunction perturbs the generation and/or the migration of neurons causing neurodevelopmental disease.
Methods
Cerebral Organoid Generation, iPSC culturing, CRISPR-Cas9 genome engineering, histological analysis, single cell sequencing, imaging.
References
Leca, I., A. Phillips, I. Hofer, L. Landler, L. Ushakova, T. D. Cushion, G. Durnberger, K. Stejskal, K. Mechtler and Keays, D. A. (2020). A proteomic survey of microtubule-associated proteins in a R402H TUBA1A mutant mouse. PLoS Genetics 16(11): e1009104.2.
Gstrein T, Edwards A, Přistoupilová A, Leca I, Breuss M, Pilat-Carotta S, Hansen AH, Tripathy R, Traunbauer AK, Hochstoeger T, Rosoklija G, Repic M, Landler L, Stránecký V, Dürnberger G, Keane TM, Zuber J, Adams DJ, Flint J, Honzik T, Gut M, Beltran S, Mechtler K, Sherr E, Kmoch S, Gut I, Keays DA. (2018). Mutations in Vps15 perturb neuronal migration in mice and are associated with neurodevelopmental disease in humans. Nature Neuroscience. Feb;21(2):207-217. doi:10.1038/s41593-017-0053-5.3.
Breuss, M., Heng, JI., Poirier, K., Tian, G., Jaglin, XH., Qu, Z., Braun, A., Gstrein, T., Ngo, L., Haas, M., Bahi-Buisson, N., Moutard, ML., Passemard, S., Verloes, A., Gressens, P., Xie, Y., Robson, KJ., Rani, DS., Thangaraj, K., Clausen, T., Chelly, J., Cowan, NJ., Keays, DA. (2012). Mutations in the β-tubulin gene TUBB5 cause microcephaly with structural brain abnormalities. Cell Reports, 2(6), 1554-1562. doi:10.1016/j.celrep.2012.11.0174.
Jaglin XH, Poirier K, Saillour Y, Buhler E, Tian G, Bahi-Buisson N, Fallet-Bianco C, Phan-Dinh-Tuy F, Kong XP, Bomont P, Castelnau-Ptakhine L, Odent S, Loget P, Kossorotoff M, Snoeck I, Plessis G, Parent P, Beldjord C, Cardoso C, Represa A, Flint J, Keays DA, Cowan NJ, Chelly J. Mutations in the beta-tubulin gene TUBB2B result in asymmetrical polymicrogyria. Nature Genetics, 41(6), 746-752. doi:10.1038/ng.380.5.
Keays DA, Tian G, Poirier K, Huang G, Siebold S, Cleak J, Oliver P, Washbourne R, Fray M, Harvey RJ, Molnar Z, Pinon M, Dear N, Brown SD, Rawlins JP, Davies KE, Cowan NJ, Patrick Nolan P, Chelly J, Flint J. Mutations in α-tubulin cause defects in neuronal migration in mice and lissencephaly in humans. Cell. 2007 Jan 12;128(1):45-57.
For further information please contact: keays@bio.lmu.de