Abstract
Ongoing, first-in-human clinical trials illustrate the feasibility and translational potential of human pluripotent stem cell (hPSC)-based cell therapies in Parkinson’s 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-NFκB 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.
Highlights
In vivo CRISPR-Cas9 screen identifies p53 limiting survival of grafted human dopamine neurons.
TNFα-NFκB pathway mediates p53-dependent human dopamine neuron death
Cell surface marker screen to enrich human dopamine neurons for translational use.
FDA approved TNF-alpha inhibitor rescues in vivo dopamine neuron survival with in vivo function.
Competing Interest Statement
L.S. is a scientific cofounder and paid consultant of BlueRock Therapeutics Inc, and a scientific cofounder of DaCapo Brainscience. T.W.K. and S.Y.K. are listed as inventors on a patent application filed and owned by the Memorial Sloan Kettering Center on the technologies described here to promote dopamine neuron survival and to enable dopamine neuron purification. The other authors declare no competing interests.
