RT Journal Article
SR Electronic
T1 Establishing cell-intrinsic limitations in cell cycle progression controls graft growth and promotes differentiation of pancreatic endocrine cells
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.03.13.990812
DO 10.1101/2020.03.13.990812
A1 Lina Sui
A1 Yurong Xin
A1 Daniela Georgieva
A1 Giacomo Diedenhofen
A1 Leena Haataja
A1 Qi Su
A1 Yong Wang
A1 Michael Zuccaro
A1 Jinrang Kim
A1 Jiayu Fu
A1 Yuan Xing
A1 Danielle Baum
A1 Robin S. Goland
A1 Jose Oberholzer
A1 Fabrizio Barbetti
A1 Peter Arvan
A1 Sandra Kleiner
A1 Dieter Egli
YR 2020
UL http://biorxiv.org/content/early/2020/03/14/2020.03.13.990812.abstract
AB Limitations in cell proliferation are a key barrier to reprogramming differentiated cells to pluripotent stem cells, and conversely, acquiring these limitations may be important to establish the differentiated state. The pancreas, and beta cells in particular have a low proliferative potential, which limits regeneration, but how these limitations are established is largely unknown. Understanding proliferation potential is important for the safty of cell replacement therapy with cell products made from pluripotent stem cell which have unlimited proliferative potential. Here we test a novel hypothesis, that these limitations are established through limitations in S-phase progression. We used a stem cell-based system to expose differentiating stem cells to small molecules that interfere with cell cycle progression either by inducing G1 arrest, impairing S-phase entry, or S-phase completion. Upon release from these molecules, we determined growth potential, differentiation and function of insulin-producing endocrine cells both in vitro and after grafting in vivo. We found that the combination of G1 arrest with a compromised ability to complete DNA replication promoted the differentiation of pancreatic progenitor cells towards insulin-producing cells, improved the stability of the differentiated state, and protected mice from diabetes without the formation of cystic growths. Therefore, a compromised ability to enter S-phase and replicate the genome is a functionally important property of pancreatic endocrine differentiation, and can be exploited to generate insulin-producing organoids with predictable growth potential after transplantation.