Directed differentiation of EA/TEF patient-derived induced pluripotent stem cells into esophageal epithelial organoids reveal SOX2 dysregulation at the anterior foregut stage
Raad, S.; David, A.; Sagniez, M.; Orfi, Z.; Dumont, N. A.; Smith, M.; Faure, C.
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A series of well-regulated cellular and molecular events result in the compartmentalization of the anterior foregut into the esophagus and trachea. Disruption of the compartmentalization process leads to esophageal atresia/tracheoesophageal fistula (EA/TEF). Therefore, the objective is to differentiate pluripotent stem cells (PSCs), namely, embryonic stem cells and iPSCs from healthy individuals and iPSCs from EA/TEF type C patients, into mature 3-dimensional esophageal organoids expressing Involucrin, Keratin-4, -13, and p63. CXCR4, SOX17, and GATA4 expression was similar in both patient and healthy endodermal cells. Key transcription factor SOX2 was significantly lower in patient-derived anterior foregut. RNA sequencing revealed critical genes GSTM1 and RAB37 to be significantly lower in patient-derived anterior foregut. Furthermore, we observed an abnormal expression of NKX2.1 in the patient-derived mature esophageal organoids. We therefore hypothesize that a transient dysregulation of SOX2 and the abnormal expression of NKX2.1 in patient-derived cells could be responsible for the abnormal foregut compartmentalization.
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