Investigating GPR161 as receptor of secreted PDZD2 in the negative regulation of Hedgehog signaling during early development

Dr Yao, Kwok Ming   (Principal Investigator (PI))

Professor Chan Danny   (Co-Investigator)

Professor Cheung Martin Chi Hang   (Co-Investigator)

42

2020-07-01

2023-12-31

1195542

Investigating GPR161 as receptor of secreted PDZD2 in the negative regulation of Hedgehog signaling during early development

early development, GLI, GPR161, Hedgehog signaling, Secreted PDZD2

Molecular BiologyDevelopmental Biology

Biology and Medicine (M)

17106620

General Research Fund (GRF)

2020

Completed

1 To test the functional relevance of the interaction between sPDZD2 and GPR161 in mouse embryonic fibroblasts (MEFs) and chick neural tube. Using a MEF line that carries a Gli-luciferase reporter, the counteracting effect of sPDZD2 on HH signaling will be investigated with and without knockdown of GPR161 expression. To investigate whether sPDZD2 exerts its effect mainly on HH signaling, RNA-seq analysis will be conducted on MEFs treated with and without sPDZD2, before and after GPR161 knockdown. Bioinformatics analysis will reveal the regulatory effects of sPDZD2 on HH targets as well as non-HH targets, and the involvement of GPR161 as receptor. Moreover, the critical domains/amino acids within GPR161 that mediate the sPDZD2-GPR161 interaction will be defined by deletion and site-directed mutagenesis and their functional effects analyzed by co-expression with sPDZD2 in chick neural tube. 2 To study the physiological relevance of the sPDZD2-GPR161 crosstalk in vivo in mutant mice. Double heterozygous Pdzd2+/-; Gpr161+/- embryos will be generated to detect aggravation of phenotypes in neural tube, limb bud and lens during early mouse development.