Abstract
Purpose :
Given that recent studies indicate Wnt5a plays a critical role in embryonic development, this study set out to investigate whether Wnt5a-activated noncanonical Wnt pathway could promote the lens embryonic differentiation.
Methods :
A “three-stage” protocol was used for lens differentiation from human embryonic stem cells(hESCs)in vitro. Exogenous addition and RNA interference were used to regulate the expression level of Wnt5a in our induction system. Quantities and size of obtained lentoid bodies were measured. QRT-PCR, western blot analysis and immunofluorescence were used for detection of gene expression.
Results :
There was a significantly increase in the quantities and sizes of generated lentoid bodies after exogenous addition of Wnt5a. Besides, the expression levels of lens specific genes including CRYAA, CRYAB, BFSP1, MIP and lens fiber differentiation regulator PROX1 were prominently increased. We further found that noncanonical Wnt signaling was activated via upregulation of DVL2, RAC1 and JNK. Conversely, when Wnt5a-knocked-down hESCs were applied for lentoid bodies induction, less and smaller lentoid bodies were harvested, decreased specific genes expression of lentoid bodies and the down-regulated activity of noncanonical Wnt/JNK pathway were observed.
Conclusions :
Our study reveals that Wnt5a could promote the differentiation of hESCs to lentoid bodies through noncanonical Wnt/JNK signaling. It also provides a fundamental basis for the prevention and treatment of lens diseases.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.