CircRNA-0100 positively regulates the differentiation of cashmere goat SHF-SCs into hair follicle lineage via sequestering miR-153-3p to heighten the KLF5 expression

Abstract

Abstract. Circular RNAs (circRNAs) have stable structures, being a covalently closed loop without 5′ and 3′ free ends. They can function as “miRNA sponges” in regulating the expression of their target genes. It was thought that circRNAs are involved in the development of the secondary hair follicle (SHF) in cashmere goats. In our previous investigation, a new circRNA named circRNA-0100 was identified from the SHF of cashmere goats, but its function is unknown. In this work, we found that circRNA-0100 exhibited significantly higher expression at anagen SHF bulge than its counterpart at telogen in cashmere goats. Based on the use of both overexpression and siRNA interference assays, our data indicated that circRNA-0100 promoted the differentiation of cashmere goat SHF stem cells (SHF-SCs) into hair follicle lineage, which was evaluated by analyzing the transcriptional level changes of six indicator genes in SHF-SCs of cashmere goats. Using the RNA pull-down technique, we showed that circRNA-0100 served as “molecular sponges” of miR-153-3p in SHF-SCs. Through the use of dual-luciferase reporter assays, our data indicated that circRNA-0100 positively regulated the transcriptional expression of the KLF5 gene via the miR-153-3p-mediated pathway. Ultimately, we showed that circRNA-0100 promoted the differentiation of SHF-SCs into hair lineage, which might be achieved via sequestering miR-153-3p to heighten the KLF5 expression in SHF-SCs of cashmere goats. Our results provide novel scientific evidence for revealing the potential molecular regulatory mechanisms on the differentiation of SHF-SCs into hair lineage in cashmere goats.

N6-Methyladenosine modification (m6A) of circRNA-ZNF638 contributes to the induced activation of SHF stem cells through miR-361-5p/Wnt5a axis in cashmere goats

“…Therefore, we speculate that m 6 A-circRNA-ZNF638 may regulate the expression of miR-361-5p in SHF-stem cells. This regulatory mode is highly similar to that reported in investigation on the differentiation of cashmere goat SHF-stem cells into hair follicle lineage, where circRNA-0100 was revealed to directly bind with miR-153-3p, and negatively regulated its expression [15]. Also, in a research on invasion and metastasis of colorectal cancer, Han and colleagues found that circLONP2 bound with miR-17, and negatively regulated its expression [28].…”

“…A r t i c l e miR-361-5p, miR-140-3p, and miR-194 (Fig. 1B), which implies that m 6 A-circRNA-ZNF638 in SHFs of cashmere goats may exert its biological functions through miRNA-mediated pathways as reported in recent publications [13][14][15]. On the other hand, the four m 6 A sites within m 6 A-circRNA-ZNF638 sequence were revealed to have the same motif of GGACU, which is consistent with the m 6 A motif: RRACH (R: A/G; H: A/C/U) reported in linear RNA molecules (Fig.…”

“…Recently, several studies showed that circRNAs were essentially implicated in the differentiation of SHF-stem cells into hair follicle lineage in cashmere goats through loss-of-function experiments [13][14][15], but little information is available about the functional role of m 6 A modified circRNAs in SHF-stem cells of cashmere goats. Also, it is unclear how circRNAs epigenetically modulate the induced activation and cell-fate decisions of SHF-stem cells of cashmere goats.…”

“…Over the past few years, extensive m 6 A modifications sites are also found in many circRNA molecules that play significant roles in a variety of biological cells through m 6 A modification dependent model [11]. To date, a considering number circRNAs have been identified in skin tissue or SHFs of cashmere goat [12], and some of them are demonstrated to positively regulate the A c c e p t e d A r t i c l e differentiation of SHF stem cells into hair follicle lineage, such as circRNA-1926 [13], circRNA-1967 [14] and circRNA-0100 [15]. Moreover, some circRNAs in SHFs of cashmere goats are verified to contain multiple m 6 A modification sites [2].…”

Construction and functional analysis of ceRNA regulatory network related to the development of secondary hair follicles in Inner Mongolia cashmere goats

“…After identifying the differentially expressed circRNAs in each comparison group, combined with the morphological changes of cashmere goat secondary hair follicles in different periods, circRNAs related to the development of secondary hair follicles were screened. In recent years, researchers discovered circRNA1967, circRNA1926, and circRNA-0100 in cashmere goat secondary hair follicles and verified their regulatory roles in secondary hair follicles ( 19 , 22 , 23 ). In this study, 113 circRNAs related to the development of secondary hair follicles, including circRNA5712, circRNA2109, circRNA9127, and circRNA3691, were screened.…”