BMB Reports

Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
권호 표지
DOI QR코드

DOI QR Code

ASCL1-mediated direct reprogramming: converting ventral midbrain astrocytes into dopaminergic neurons for Parkinson's disease therapy

  • Sang Hui Yong (Graduate School of Biomedical Science and Engineering, Hanyang University) ;
  • Sang-Mi Kim (Hanyang Biomedical Research Institute, Hanyang University) ;
  • Gyeong Woon Kong (Graduate School of Biomedical Science and Engineering, Hanyang University) ;
  • Seung Hwan Ko (Graduate School of Biomedical Science and Engineering, Hanyang University) ;
  • Eun-Hye Lee (Neuroregeneration and Stem Cell Programs, Institute for Cell Engineering, Johns Hopkins University School of Medicine) ;
  • Yohan Oh (Graduate School of Biomedical Science and Engineering, Hanyang University) ;
  • Chang-Hwan Park (Graduate School of Biomedical Science and Engineering, Hanyang University)
  • Received : 2023.11.22
  • Accepted : 2024.01.23
  • Published : 2024.08.31
Download PDF
⟨ Previous Next ⟩

Abstract

Parkinson's disease (PD), characterized by dopaminergic neuron degeneration in the substantia nigra, is caused by various genetic and environmental factors. Current treatment methods are medication and surgery; however, a primary therapy has not yet been proposed. In this study, we aimed to develop a new treatment for PD that induces direct reprogramming of dopaminergic neurons (iDAN). Achaete-scute family bHLH transcription factor 1 (ASCL1) is a primary factor that initiates and regulates central nervous system development and induces neurogenesis. In addition, it interacts with BRN2 and MYT1L, which are crucial transcription factors for the direct conversion of fibroblasts into neurons. Overexpression of ASCL1 along with the transcription factors NURR1 and LMX1A can directly reprogram iDANs. Using a retrovirus, GFP-tagged ASCL1 was overexpressed in astrocytes. One week of culture in iDAN convertsion medium reprogrammed the astrocytes into iDANs. After 7 days of differentiation, TH+/TUJ1+ cells emerged. After 2 weeks, the number of mature TH+/TUJ1+ dopaminergic neurons increased. Only ventral midbrain (VM) astrocytes exhibited these results, not cortical astrocytes. Thus, VM astrocytes can undergo direct iDAN reprogramming with ASCL1 alone, in the absence of transcription factors that stimulate dopaminergic neurons development.

Keywords

Acknowledgement

This research was supported by grants from the Basic Science Research Program (NRF-2019R1A2C2005681), the K-Brain Project of the National Research Foundation funded by Ministry of Science and ICT (RS-2023-00266171), and the Korea Dementia Research Project through the Korea Dementia Research Center funded by Ministry of Health and Welfare and MSIT (HU22C0143).

References

  1. McGregor MM and Nelson AB (2019) Circuit mechanisms of Parkinson's disease. Neuron 101, 1042-1056 https://doi.org/10.1016/j.neuron.2019.03.004
  2. Group PS (2004) Levodopa and the progression of Parkinson's disease. N Engl J Med 351, 2498-2508 https://doi.org/10.1056/NEJMoa033447
  3. Fabbrini G, Brotchie JM, Grandas F, Nomoto M and Goetz CG (2007) Levodopa-induced dyskinesias. Mov Disord 22, 1379-1389 https://doi.org/10.1002/mds.21475
  4. Freed CR, Greene PE, Breeze RE et al (2001) Transplantation of embryonic dopamine neurons for severe Parkinson's disease. N Engl J Med 344, 710-719 https://doi.org/10.1056/NEJM200103083441002
  5. Bjorklund LM, Sanchez-Pernaute R, Chung S et al (2002) Embryonic stem cells develop into functional dopaminergic neurons after transplantation in a Parkinson rat model. Proc Natl Acad Sci U S A 99, 2344-2349 https://doi.org/10.1073/pnas.022438099
  6. Hallett PJ, Deleidi M, Astradsson A et al (2015) Successful function of autologous iPSC-derived dopamine neurons following transplantation in a non-human primate model of Parkinson's disease. Cell Stem Cell 16, 269-274 https://doi.org/10.1016/j.stem.2015.01.018
  7. Pfisterer U, Kirkeby A, Torper O et al (2011) Direct conversion of human fibroblasts to dopaminergic neurons. Proc Natl Acad Sci U S A 108, 10343-10348 https://doi.org/10.1073/pnas.1105135108
  8. Pang ZP, Yang N, Vierbuchen T et al (2011) Induction of human neuronal cells by defined transcription factors. Nature 476, 220-223 https://doi.org/10.1038/nature10202
  9. Siracusa R, Fusco R and Cuzzocrea S (2019) Astrocytes: role and functions in brain pathologies. Front Pharmacol 10, 1114
  10. Wapinski OL, Lee QY, Chen AC et al (2017) Rapid chromatin switch in the direct reprogramming of fibroblasts to neurons. Cell Rep 20, 3236-3247 https://doi.org/10.1016/j.celrep.2017.09.011
  11. Rao Z, Wang R, Li S et al (2021) Molecular mechanisms underlying ascl1-mediated astrocyte-to-neuron conversion. Stem Cell Reports 16, 534-547 https://doi.org/10.1016/j.stemcr.2021.01.006
  12. Chanda S, Ang CE, Davila J et al (2014) Generation of induced neuronal cells by the single reprogramming factor ASCL1. Stem Cell Reports 3, 282-296 https://doi.org/10.1016/j.stemcr.2014.05.020
  13. Sohn J (2021) Efficient neuronal direct conversion of mouse astrocytes by ASCL1 dephosphorylation. Master of Science, dissertation, Hanyang University
  14. Han JY, Lee EH, Kim SM and Park CH (2023) Efficient generation of dopaminergic neurons from mouse ventral midbrain astrocytes. Biomol Ther (Seoul) 31, 264-275 https://doi.org/10.4062/biomolther.2022.140
  15. Antoni D, Burckel H, Josset E and Noel G (2015) Three-dimensional cell culture: a breakthrough in vivo. Int J Mol Sci 16, 5517-5527 https://doi.org/10.3390/ijms16035517
  16. Zhou S, Szczesna K, Ochalek A et al (2016) Neurosphere based differentiation of human iPSC improves astrocyte differentiation. Stem Cells Int 2016, 4937689
  17. Morel L, Chiang MSR, Higashimori H et al (2017) Molecular and functional properties of regional astrocytes in the adult brain. J Neurosci 37, 8706-8717 https://doi.org/10.1523/JNEUROSCI.3956-16.2017
  18. Song JJ, Oh SM, Kwon OC et al (2021) Cografting astrocytes improves cell therapeutic outcomes in a Parkinson's disease model. J Clin Invest 128, 463-482 https://doi.org/10.1172/JCI93924
  19. Reynolds BA, Tetzlaff W and Weiss S (1992) A multipotent EGF-responsive striatal embryonic progenitor cell produces neurons and astrocytes. J Neurosci 12, 4565-4574 https://doi.org/10.1523/JNEUROSCI.12-11-04565.1992
  20. Qian X, Song H and Ming Gl (2019) Brain organoids: advances, applications and challenges. Development 146, dev166074
  21. Lancaster MA and Knoblich JA (2014) Generation of cerebral organoids from human pluripotent stem cells. Nature Protoc 9, 2329-2340 https://doi.org/10.1038/nprot.2014.158
  22. Quadrato G, Nguyen T, Macosko EZ et al (2017) Cell diversity and network dynamics in photosensitive human brain organoids. Nature 545, 48-53 https://doi.org/10.1038/nature22047
  23. Lee JH, Shaker MR, Park SH, Sun W (2023) Transcriptional signature of valproic acid-induced neural tube defects in human spinal cord organoids. Int J Stem Cells 16, 385-393 https://doi.org/10.15283/ijsc23012
  24. Wang LL, Serrano C, Zhong X, Ma S, Zou Y and Zhang CL (2021) Revisiting astrocyte to neuron conversion with lineage tracing in vivo. Cell 184, 5465-5481 e5416