iPSC-derived NF1-CDKN2A -PRC2 deficient neural crest cells mimic glial-to-neuro-mesenchymal transition and form MPNST-like tumors in vivo

Neurofibromatosis Type 1 (NF1) predisposes to peripheral nerve tumor development. Commonly, the progression from a benign plexiform neurofibroma (PNF) towards a deadly malignant peripheral nerve sheath tumor (MPNST) involves a poorly understood glial-to-mesenchymal transition and the sequential loss of NF1, CDKN2A , and polycomb repressive complex 2 (PRC2). Using an iPSC-derived neural crest (NC) model, we reproduced this malignant transformation. NF1-CDKN2A double-knockout (2KO) NCs retained glial differentiation capacity and generated neurofibroma-like tumors in vivo , requiring inactivation of p14ARF and p16INK4A. Additional PRC2 loss (3KO) disrupted pluripotency and induced mesenchymal stem cell-like features. 3KO NC globally reassembled chromatin accessibility, impeding gliogenesis by SOX10 epigenetic silencing and activating neuro-mesenchymal programs by chromatin opening, reinforced in MPNSTs. This glial-to-neuro-mesenchymal transition recapitulated neurofibroma-to-MPNST progression, since 3KO NC spheres formed MPNST-like tumors in vivo , genuinely mimicking early-stage MPNST development. This iPSC-based model constitutes a platform for studying the molecular pathogenesis of NF1-associated sarcomagenesis and therapeutic discovery.