Abstract
Parkinson's disease (PD)-specific neurons, grown in standard 2D cultures, typically only display weak endophenotypes. The cultivation of PD patient-specific neurons, derived from induced pluripotent stem cells carrying the LRRK2-G2019S mutation, is optimized in 3D microfluidics. The automated image analysis algorithms are implemented to enable pharmacophenomics in disease-relevant conditions. In contrast to 2D cultures, this 3D approach reveals robust endophenotypes. High-content imaging data show decreased dopaminergic differentiation and branching complexity, altered mitochondrial morphology, and increased cell death in LRRK2-G2019S neurons compared to isogenic lines without using stressor agents. Treatment with the LRRK2 inhibitor 2 (Inh2) rescues LRRK2-G2019S-dependent dopaminergic phenotypes. Strikingly, a holistic analysis of all studied features shows that the genetic background of the PD patients, and not the LRRK2-G2019S mutation, constitutes the strongest contribution to the phenotypes. These data support the use of advanced in vitro models for future patient stratification and personalized drug development.
| Original language | English |
|---|---|
| Article number | 1800927 |
| Journal | Advanced Science |
| Volume | 6 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 09.01.2019 |
Funding
The authors thank Prof. Dr. Hans R. Schöler from the Max-Planck-Gesellschaft, Prof. Dr. Thomas Gasser from the Universitatsklinikum Tuebingen, and Dr. Jared Sterneckert from the CRTD for providing us with cell lines. Funding: The JCS lab is supported by the Fonds National de la Recherche (FNR) (CORE, C13/BM/5791363 and Proof-of-Concept program PoC15/11180855 & PoC16/11559169). This is an EU Joint Programme—Neurodegenerative Disease Research (JPND) project (INTER/JPND/14/02; INTER/JPND/15/11092422). Further support came from the SysMedPD project, which has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement no, 668738. J.J., E.L.M., and X.Q. were supported by fellowships from the FNR (AFR, Aides à la Formation-Recherche). Finally, the authors also thank the private donors who support the work at the Luxembourg Centre for Systems Biomedicine. E.L.M. was also supported by an Aides à la Formation-Recherche training allowance from FNR. The AG lab is supported by grants from the FNR in the ATTRACT (Model IPD, 9631103) and INTER programs (ProtectMove, 11250962). Moreover, AG receives funding from the German Research Foundation within the framework of the Research Unit “ProtectMove” (GR 3731/5-1; FOR 2488/1). E.G. acknowledges support by the Fonds Nationale de la Recherche (FNR) Luxembourg through the National Centre of Excellence in Research (NCER) on Parkinson’s disease (I1R-BIC-PFN-15NCER). E.G. performed microarray differential expression and heat map analyses. J.C.S. conceived the project. S.B., M.F., K.W., N.O., and S.L.N. performed the experiments. S.B., A.G., and J.C.S. designed the experiments. S.B. and J.C.S. wrote the paper. X.Q. introduced the LRRK2-G2019S mutation in the Healthy2 line. J.W. generated the PD1 line. P.A. wrote and conceived the algorithms for image analysis. J.J. contributed to the image analysis optimization and performed the statistical analysis in R. E.G. performed microarray differential expression and heat map analyses. E.L.M. contributed to the optimization of the microfluidic culture system. R.M.T.F. provided critical reading and scientific discussion. J.S. performed Zn analysis. L.S. performed the ROC and AUC analysis.
Research Areas and Centers
- Academic Focus: Center for Brain, Behavior and Metabolism (CBBM)
