TNF-NFkB-p53 axis restricts in vivo survival of hPSC-derived dopamine neuron
Ongoing, first-in-human clinical trials illustrate the feasibility and translational potential of human pluripotent stem cell (hPSC)-based cell therapies in Parkinsons disease (PD). However, a major unresolved challenge in the field is the extensive cell death following transplantation with <10% of grafted dopamine neurons surviving. Here, we performed a pooled CRISPR/Cas9 screen to enhance survival of postmitotic dopamine neurons in vivo. We identified p53-mediated apoptotic cell death as major contributor to dopamine neuron loss and uncovered a causal link of TNFa-NF{kappa}B signaling in limiting cell survival. As a translationally applicable strategy to purify postmitotic dopamine neurons, we performed a cell surface marker screen that enabled purification without the need for genetic reporters. Combining cell sorting with adalimumab pretreatment, a clinically approved and widely used TNFa inhibitor, enabled efficient engraftment of postmitotic dopamine neurons leading to extensive re-innervation and functional recovery in a preclinical PD mouse model. Thus, transient TNFa inhibition presents a clinically relevant strategy to enhance survival and enable engraftment of postmitotic human PSC-derived dopamine neurons in PD. HighlightsO_LIIn vivo CRISPR-Cas9 screen identifies p53 limiting survival of grafted human dopamine neurons. C_LIO_LITNF-NF{kappa}B pathway mediates p53-dependent human dopamine neuron death C_LIO_LICell surface marker screen to enrich human dopamine neurons for translational use. C_LIO_LIFDA approved TNF-alpha inhibitor rescues in vivo dopamine neuron survival with in vivo function. C_LI