Ongoing, early-stage clinical trials illustrate the translational potential of human pluripotent stem cell (hPSC)-based cell therapies in Parkinson’s disease (PD). However, an unresolved challenge is the extensive cell death following transplantation. Here, we performed a pooled CRISPR-Cas9 screen to enhance postmitotic dopamine neuron survival in vivo. We identified p53-mediated apoptotic cell death as a major contributor to dopamine neuron loss and uncovered a causal link of tumor necrosis factor alpha (TNF-α)-nuclear factor κB (NF-κB) signaling in limiting cell survival. As a translationally relevant strategy to purify postmitotic dopamine neurons, we identified cell surface markers that enable purification without the need for genetic reporters. Combining cell sorting and treatment with adalimumab, a clinically approved TNF-α inhibitor, enabled efficient engraftment of postmitotic dopamine neurons with extensive reinnervation and functional recovery in a preclinical PD mouse model. Thus, transient TNF-α inhibition presents a clinically relevant strategy to enhance survival and enable engraftment of postmitotic hPSC-derived dopamine neurons in PD.

正在进行的早期临床试验说明了基于人类多能干细胞 (hPSC) 的细胞疗法在帕金森病 (PD) 中的转化潜力。然而，一个尚未解决的挑战是移植后广泛的细胞死亡。在这里，我们进行了 CRISPR-Cas9 合并筛选，以提高有丝分裂后多巴胺神经元在体内的存活率。我们确定 p53 介导的细胞凋亡是多巴胺神经元丢失的主要原因，并发现肿瘤坏死因子 α (TNF-α)-核因子 κB (NF-κB) 信号传导在限制细胞存活中的因果关系。作为纯化有丝分裂后多巴胺神经元的翻译相关策略，我们鉴定了无需基因报告基因即可进行纯化的细胞表面标记。将细胞分选和阿达木单抗（一种临床批准的 TNF-α 抑制剂）治疗相结合，能够在临床前 PD 小鼠模型中有效植入有丝分裂后多巴胺神经元，并进行广泛的神经支配和功能恢复。因此，短暂的 TNF-α 抑制提供了一种临床相关策略，可提高 PD 中的存活率并实现有丝分裂后 hPSC 衍生的多巴胺神经元的植入。