Single-cell Transcriptomics Reveals Correct Developmental Dynamics and High-quality Midbrain Cell Types by Improved HESC Differentiation

Overview

Journal Stem Cell Reports

Publisher Cell Press

Specialty Cell Biology

Date 2022 Nov 18

PMID 36400027

Authors

Kaneyasu Nishimura

Shanzheng Yang

Ka Wai Lee

Emilia Sif Asgrimsdottir

Kasra Nikouei

Wojciech Paslawski

Sabine Gnodde

Guochang Lyu

Lijuan Hu

Carmen Salto

Per Svenningsson

Jens Hjerling-Leffler

Sten Linnarsson

Ernest Arenas

Affiliations

Soon will be listed here.

Abstract

Stem cell technologies provide new opportunities for modeling cells in health and disease and for regenerative medicine. In both cases, developmental knowledge and defining the molecular properties and quality of the cell types is essential. In this study, we identify developmental factors important for the differentiation of human embryonic stem cells (hESCs) into functional midbrain dopaminergic (mDA) neurons. We found that laminin-511, and dual canonical and non-canonical WNT activation followed by GSK3β inhibition plus FGF8b, improved midbrain patterning. In addition, neurogenesis and differentiation were enhanced by activation of liver X receptors and inhibition of fibroblast growth factor signaling. Moreover, single-cell RNA-sequencing analysis revealed a developmental dynamics similar to that of the endogenous human ventral midbrain and the emergence of high-quality molecularly defined midbrain cell types, including mDA neurons. Our study identifies novel factors important for human midbrain development and opens the door for a future application of molecularly defined hESC-derived cell types in Parkinson disease.

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References

Thomson J, Itskovitz-Eldor J, Shapiro S, Waknitz M, Swiergiel J, Marshall V . Embryonic stem cell lines derived from human blastocysts. Science. 1998; 282(5391):1145-7. DOI: 10.1126/science.282.5391.1145. View

Rodin S, Antonsson L, Niaudet C, Simonson O, Salmela E, Hansson E . Clonal culturing of human embryonic stem cells on laminin-521/E-cadherin matrix in defined and xeno-free environment. Nat Commun. 2014; 5:3195. DOI: 10.1038/ncomms4195. View

Tiklova K, Bjorklund A, Lahti L, Fiorenzano A, Nolbrant S, Gillberg L . Single-cell RNA sequencing reveals midbrain dopamine neuron diversity emerging during mouse brain development. Nat Commun. 2019; 10(1):581. PMC: 6362095. DOI: 10.1038/s41467-019-08453-1. View

Koch P, Opitz T, Steinbeck J, Ladewig J, Brustle O . A rosette-type, self-renewing human ES cell-derived neural stem cell with potential for in vitro instruction and synaptic integration. Proc Natl Acad Sci U S A. 2009; 106(9):3225-30. PMC: 2651316. DOI: 10.1073/pnas.0808387106. View

Arenas E . Wnt signaling in midbrain dopaminergic neuron development and regenerative medicine for Parkinson's disease. J Mol Cell Biol. 2014; 6(1):42-53. DOI: 10.1093/jmcb/mju001. View

Lees A, Hardy J, Revesz T . Parkinson's disease. Lancet. 2009; 373(9680):2055-66. DOI: 10.1016/S0140-6736(09)60492-X. View

Ferri A, Lin W, Mavromatakis Y, Wang J, Sasaki H, Whitsett J . Foxa1 and Foxa2 regulate multiple phases of midbrain dopaminergic neuron development in a dosage-dependent manner. Development. 2007; 134(15):2761-9. DOI: 10.1242/dev.000141. View

Kim T, Piao J, Koo S, Kriks S, Chung S, Betel D . Biphasic Activation of WNT Signaling Facilitates the Derivation of Midbrain Dopamine Neurons from hESCs for Translational Use. Cell Stem Cell. 2021; 28(2):343-355.e5. PMC: 8006469. DOI: 10.1016/j.stem.2021.01.005. View

Chambers S, Fasano C, Papapetrou E, Tomishima M, Sadelain M, Studer L . Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nat Biotechnol. 2009; 27(3):275-80. PMC: 2756723. DOI: 10.1038/nbt.1529. View

10.

Villaescusa J, Li B, Toledo E, Rivetti di Val Cervo P, Yang S, Stott S . A PBX1 transcriptional network controls dopaminergic neuron development and is impaired in Parkinson's disease. EMBO J. 2016; 35(18):1963-78. PMC: 5282836. DOI: 10.15252/embj.201593725. View

11.

Andersson E, Salto C, Villaescusa J, cajanek L, Yang S, Bryjova L . Wnt5a cooperates with canonical Wnts to generate midbrain dopaminergic neurons in vivo and in stem cells. Proc Natl Acad Sci U S A. 2013; 110(7):E602-10. PMC: 3574960. DOI: 10.1073/pnas.1208524110. View

12.

Toledo E, Yang S, Gyllborg D, van Wijk K, Sinha I, Varas-Godoy M . Srebf1 Controls Midbrain Dopaminergic Neurogenesis. Cell Rep. 2020; 31(5):107601. DOI: 10.1016/j.celrep.2020.107601. View

13.

Kirkeby A, Grealish S, Wolf D, Nelander J, Wood J, Lundblad M . Generation of regionally specified neural progenitors and functional neurons from human embryonic stem cells under defined conditions. Cell Rep. 2012; 1(6):703-14. DOI: 10.1016/j.celrep.2012.04.009. View

14.

Li W, Englund E, Widner H, Mattsson B, van Westen D, Latt J . Extensive graft-derived dopaminergic innervation is maintained 24 years after transplantation in the degenerating parkinsonian brain. Proc Natl Acad Sci U S A. 2016; 113(23):6544-9. PMC: 4988567. DOI: 10.1073/pnas.1605245113. View

15.

Theofilopoulos S, Wang Y, Kitambi S, Sacchetti P, Sousa K, Bodin K . Brain endogenous liver X receptor ligands selectively promote midbrain neurogenesis. Nat Chem Biol. 2013; 9(2):126-33. DOI: 10.1038/nchembio.1156. View

16.

Castelo-Branco G, Wagner J, Rodriguez F, Kele J, Sousa K, Rawal N . Differential regulation of midbrain dopaminergic neuron development by Wnt-1, Wnt-3a, and Wnt-5a. Proc Natl Acad Sci U S A. 2003; 100(22):12747-52. PMC: 240689. DOI: 10.1073/pnas.1534900100. View

17.

Simon H, SAUERESSIG H, Wurst W, Goulding M, OLeary D . Fate of midbrain dopaminergic neurons controlled by the engrailed genes. J Neurosci. 2001; 21(9):3126-34. PMC: 6762576. View

18.

Arenas E, Denham M, Villaescusa J . How to make a midbrain dopaminergic neuron. Development. 2015; 142(11):1918-36. DOI: 10.1242/dev.097394. View

19.

Andersson E, Tryggvason U, Deng Q, Friling S, Alekseenko Z, Robert B . Identification of intrinsic determinants of midbrain dopamine neurons. Cell. 2006; 124(2):393-405. DOI: 10.1016/j.cell.2005.10.037. View

20.

Nunes I, Tovmasian L, Silva R, Burke R, Goff S . Pitx3 is required for development of substantia nigra dopaminergic neurons. Proc Natl Acad Sci U S A. 2003; 100(7):4245-50. PMC: 153078. DOI: 10.1073/pnas.0230529100. View