(317b) Skin Tissue Derived Neural Crest Stem Cells and Metabolic Requirements for Multipotency

Neural crest (NC) cells are a multipotent stem cell population that give rise to a diverse array of cells in the body, including peripheral neurons, Schwann cells (SC), smooth muscle cells, and melanocytes. Neural crest (NC) induction during embryogenesis takes place in response to two key signaling events: the Wnt pathway that is active at the neural plate border; and BMP signaling from the developing mesoderm. In our lab, we have isolated NC-like stem cells (KC-NCs) from adult skin tissue, and have shown that these resemble the NCs that originate during development, including their differentiation potential. Hence, these cells serve as a safe and efficient patient-specific source for cell therapy. However, KC-NCs are very transient, and lose their stem cell potential over time in culture limiting their clinical use. Methods to find ways to preserve the multipotency for these cells is thus crucial to exploit their regenerative potential.

In this regard, we mimicked the developmental microenvironment during NC induction and treated the cells with BMP2 and the Wnt agonist CHIR99021 to preserve multipotency of KC-NCs. Combinatorial Wnt/BMP2 signaling sustained the expression of transient NC genes including Sox10, Pax7, AP2A, HNK1 and TrkC in culture. Subsequently, this treatment enhanced the differentiation of KC-NCs to melanocytes, Schwann cells and smooth muscle cells, as indicated by increased expression of MITF, KROX20 and αSMA respectively. We discovered that Wnt/BMP signaling rewired the metabolic landscape of NCSCs to confer multipotency. This was achieved by activating anaerobic glycolysis in KC-NCs, which increased lactate production, glucose consumption and sensitivity to insulin stimulation. Energy demands of multipotent KC-NCs were evaluated by employing the Seahorse ATP Rate assay. Indeed, cells treated with CHIR/BMP2 derived most of their ATP from glycolysis, and the mitochondrial activity was diminished. Taken, together, this study elucidates how developmental events control the carbon metabolism and bioenergetics state of NCs, which is important in understanding metabolic disorders arising from dysregulation of NC induction and differentiation during embryogenesis. We also describe a simple method using a cocktail of chemicals and growth factors for maintaining KC-NC multipotency for treatment of demyelinating and other diseases.