Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW) and Department of Neuroscience, University of Copenhagen, 2200 Copenhagen N, Denmark.
Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW) and Department of Neuroscience, University of Copenhagen, 2200 Copenhagen N, Denmark; Department of Experimental Medical Science, Lund University, 22184 Lund, Sweden; Wallenberg Center for Molecular Medicine, Department of Experimental Medical Sciences, Lund University, 22184 Lund, Sweden.
Stem Cell Reports. 2024 Jun 11;19(6):830-838. doi: 10.1016/j.stemcr.2024.04.010. Epub 2024 May 16.
The differentiation of human pluripotent stem cells into ventral mesencephalic dopaminergic (DA) fate is relevant for the treatment of Parkinson's disease. Shortcuts to obtaining DA cells through direct reprogramming often include forced expression of the transcription factor LMX1A. Although reprogramming with LMX1A can generate tyrosine hydroxylase (TH)-positive cells, their regional identity remains elusive. Using an in vitro model of early human neural tube patterning, we report that forced LMX1A expression induced a ventral-to-dorsal fate shift along the entire neuroaxis with the emergence of roof plate fates despite the presence of ventralizing molecules. The LMX1A-expressing progenitors gave rise to grafts containing roof plate-derived choroid plexus cysts as well as ectopically induced TH-positive neurons of a forebrain identity. Early activation of LMX1A prior to floor plate specification was necessary for the dorsalizing effect. Our work suggests using caution in employing LMX1A for the induction of DA fate, as this factor may generate roof plate rather than midbrain fates.
人类多能干细胞向腹侧中脑多巴胺能（DA）命运的分化与帕金森病的治疗有关。通过直接重编程获得 DA 细胞的捷径通常包括转录因子 LMX1A 的强制表达。尽管 LMX1A 的重编程可以产生酪氨酸羟化酶（TH）阳性细胞，但它们的区域身份仍然难以捉摸。使用早期人神经管模式形成的体外模型，我们报告说，尽管存在中胚层化分子，但强制表达 LMX1A 会诱导整个神经轴的腹侧到背侧命运转变，出现顶板命运。表达 LMX1A 的祖细胞产生了含有顶板衍生脉络丛囊肿的移植物，以及异位诱导的具有前脑特征的 TH 阳性神经元。在底板特化之前早期激活 LMX1A 对于背侧化效应是必要的。我们的工作表明，在诱导 DA 命运时应谨慎使用 LMX1A，因为该因子可能产生顶板而不是中脑命运。
