Optogenetic manipulation of BMP signaling to drive chondrogenic differentiation of hPSCs
Paul E.A. Humphreys,
Steven Woods,
Nicola Bates,
Kirsty M. Rooney,
Fabrizio E. Mancini,
Cerys Barclay,
Julieta O’Flaherty,
Franck P. Martial,
Marco A.N. Domingos,
Susan J. Kimber
Affiliations
Paul E.A. Humphreys
Division of Cell Matrix & Regenerative Medicine, School of Biological Sciences, Faculty of Biology Medicine and Health, University of Manchester, Oxford Road, Manchester M13 9PT, UK
Steven Woods
Division of Cell Matrix & Regenerative Medicine, School of Biological Sciences, Faculty of Biology Medicine and Health, University of Manchester, Oxford Road, Manchester M13 9PT, UK
Nicola Bates
Division of Cell Matrix & Regenerative Medicine, School of Biological Sciences, Faculty of Biology Medicine and Health, University of Manchester, Oxford Road, Manchester M13 9PT, UK
Kirsty M. Rooney
Division of Cell Matrix & Regenerative Medicine, School of Biological Sciences, Faculty of Biology Medicine and Health, University of Manchester, Oxford Road, Manchester M13 9PT, UK
Fabrizio E. Mancini
Division of Cell Matrix & Regenerative Medicine, School of Biological Sciences, Faculty of Biology Medicine and Health, University of Manchester, Oxford Road, Manchester M13 9PT, UK; Department of Mechanical, Aerospace, and Civil Engineering, Faculty of Science and Engineering, University of Manchester, Oxford Road, Manchester M13 9PT, UK
Cerys Barclay
Division of Cell Matrix & Regenerative Medicine, School of Biological Sciences, Faculty of Biology Medicine and Health, University of Manchester, Oxford Road, Manchester M13 9PT, UK
Julieta O’Flaherty
Division of Cell Matrix & Regenerative Medicine, School of Biological Sciences, Faculty of Biology Medicine and Health, University of Manchester, Oxford Road, Manchester M13 9PT, UK
Franck P. Martial
Division of Neuroscience & Experimental Psychology, Faculty of Biology, Medicine and Health, University of Manchester, Oxford Road, Manchester M13 9PT, UK
Marco A.N. Domingos
Department of Mechanical, Aerospace, and Civil Engineering, Faculty of Science and Engineering, University of Manchester, Oxford Road, Manchester M13 9PT, UK
Susan J. Kimber
Division of Cell Matrix & Regenerative Medicine, School of Biological Sciences, Faculty of Biology Medicine and Health, University of Manchester, Oxford Road, Manchester M13 9PT, UK; Corresponding author
Summary: Optogenetics is a rapidly advancing technology combining photochemical, optical, and synthetic biology to control cellular behavior. Together, sensitive light-responsive optogenetic tools and human pluripotent stem cell differentiation models have the potential to fine-tune differentiation and unpick the processes by which cell specification and tissue patterning are controlled by morphogens. We used an optogenetic bone morphogenetic protein (BMP) signaling system (optoBMP) to drive chondrogenic differentiation of human embryonic stem cells (hESCs). We engineered light-sensitive hESCs through CRISPR-Cas9-mediated integration of the optoBMP system into the AAVS1 locus. The activation of optoBMP with blue light, in lieu of BMP growth factors, resulted in the activation of BMP signaling mechanisms and upregulation of a chondrogenic phenotype, with significant transcriptional differences compared to cells in the dark. Furthermore, cells differentiated with light could form chondrogenic pellets consisting of a hyaline-like cartilaginous matrix. Our findings indicate the applicability of optogenetics for understanding human development and tissue engineering.
