AUTHOR=Xue Shanshan , Du Xuanjin , Yu Minghui , Ju Haixin , Tan Lihong , Li Yaxin , Liu Jialu , Wang Chunyan , Wu Xiaohui , Xu Hong , Shen Qian 

TITLE=Overexpression of long noncoding RNA 4933425B07Rik leads to renal hypoplasia by inactivating Wnt/β-catenin signaling pathway

JOURNAL=Frontiers in Cell and Developmental Biology

VOLUME=Volume 11 - 2023

YEAR=2023

URL=https://www.frontiersin.org/journals/cell-and-developmental-biology/articles/10.3389/fcell.2023.1267440

DOI=10.3389/fcell.2023.1267440

ISSN=2296-634X

ABSTRACT=<p>Congenital anomalies of the kidney and urinary tract (CAKUT) is a general term for a class of diseases that are mostly caused by intrauterine genetic development limitation. Without timely intervention, certain children with CAKUT may experience progressive decompensation and a rapid decline in renal function, which will ultimately result in end-stage renal disease. At present, a comprehensive understanding of the pathogenic signaling events of CAKUT is lacking. The role of long noncoding RNAs (lncRNAs) in renal development and disease have recently received much interest. In previous research, we discovered that mice overexpressing the lncRNA <italic>4933425B07Rik</italic> (<italic>Rik</italic>) showed a range of CAKUT phenotypes, primarily renal hypoplasia. The current study investigated the molecular basis of renal hypoplasia caused by <italic>Rik</italic> overexpression. We first used Rapid Amplification of cDNA ends (RACE) to obtain the full-length sequence of <italic>Rik</italic> in <italic>Rik</italic><sup><italic>+/+</italic></sup>;<italic>Hoxb7</italic> mice. Mouse proximal renal tubule epithelial cells (MPTCs) line with <italic>Rik</italic> overexpression was constructed using lentiviral methods, and mouse metanephric mesenchyme cell line (MK3) with <italic>Rik</italic> knockout was then constructed by the CRISPR‒Cas9 method. We performed RNA-seq on the <italic>Rik</italic>-overexpressing cell line to explore possible differentially expressed molecules and pathways. mRNA expression was confirmed by qRT‒PCR. Reduced levels of <italic>Wnt10b</italic>, <italic>Fzd8</italic>, and <italic>β-catenin</italic> were observed when <italic>Rik</italic> was expressed robustly. On the other hand, these genes were more highly expressed when <italic>Rik</italic> was knocked out. These results imply that overabundance of <italic>Rik</italic> might inhibit the Wnt/β-catenin signaling pathway, which may result in renal hypoplasia. In general, such research might help shed light on CAKUT causes and processes and offer guidance for creating new prophylactic and therapeutic strategies.</p>