Trajectory analysis of hepatic stellate cell differentiation reveals metabolic regulation of cell commitment and fibrosis
Raquel A. Martínez García de la Torre, Júlia Vallverdú, Zhixin Xu, Sílvia Ariño, Raquel Ferrer‐Lorente, Laura Zanatto, María Mercado-Gómez, Beatriz Aguilar‐Bravo, Paloma Ruiz-Blázquez, M. Guadalupe Fernández, Artur Navarro‐Gascon, Albert Blasco‐Roset, Paula Sànchez‐Fernàndez‐de‐Landa, Joan Pera, Damià Romero–Moya, Paula Ayuso Garcia, Celia Martínez–Sánchez, Laura Sererols Viñas, Paula Cantallops Vilà, Carmen I. Cárcamo Giráldez, Andrew McQuillin, Marsha Y. Morgan, Daniel Moya-Rull, Núria Montserrat, Delphine Eberlé, Bart Staels, Bénédicte Antoine, Mikel Azkargorta, Juan José Lozano, Maria Luz Martínez‐Chantar, Alessandra Giorgetti, Félix Elortza, Anna Planavila, Marta Varela‐Rey, Ashwin Woodhoo, António Zorzano, Isabel Graupera, Anna Moles, Mar Coll, Silvia Affò, Pau Sancho‐Bru
Abstract
Defining the trajectory of cells during differentiation and disease is key for uncovering the mechanisms driving cell fate and identity. However, trajectories of human cells remain largely unexplored due to the challenges of studying them with human samples. In this study, we investigate the proteome trajectory of iPSCs differentiation to hepatic stellate cells (diHSCs) and identify RORA as a key transcription factor governing the metabolic reprogramming of HSCs necessary for diHSCs’ commitment, identity, and activation. Using RORA deficient iPSCs and pharmacologic interventions, we show that RORA is required for early differentiation and prevents diHSCs activation by reducing the high energetic state of the cells. While RORA knockout mice have enhanced fibrosis, RORA agonists rescue multi-organ fibrosis in in vivo models. Notably, RORA expression correlates negatively with liver fibrosis and HSCs activation markers in patients with liver disease. This study reveals that RORA regulates cell metabolic plasticity, important for mesoderm differentiation, pericyte quiescence, and fibrosis, influencing cell commitment and disease. Here, the authors investigate the proteome trajectory of human iPSCs differentiating into hepatic stellate cells (diHSCs) and identify RORA as a key transcription factor that governs metabolic reprogramming essential for diHSCs’ commitment, identity, and quiescent phenotype.