mProX™ Human GPR65 Stable Cell Line

Li G, et al. "Intestinal epithelial pH-sensing receptor GPR65 maintains mucosal homeostasis via ." Gut microbes, 2023.
The regulation of barrier function and mucosal homeostasis by intestinal epithelial cells (IECs) is essential for maintaining a healthy gut microenvironment. However, the underlying host-derived regulatory networks that modulate intestinal antimicrobial defenses remain incompletely understood. In this study, Gpr65 (Gpr65(DeltaIEC)) mice were generated to assess the role of epithelial GPR65 in DSS- and C. rodentium-induced murine colitis models. RNA sequencing analysis was performed on colonic IECs from Gpr65(fl/fl) and Gpr65(DeltaIEC) mice, while colonoids and colonic epithelial cell lines were utilized to assess the pH-sensing effect of GPR65. The expression of GPR65 was also investigated in IECs from patients with inflammatory bowel disease (IBD) and DSS colitis mice. Our results demonstrated that, in the absence of GPR65, homeostatic antimicrobial programs, including the production of antimicrobial peptides (AMPs) and defense response-associated proteins, were abrogated in IECs. Gpr65(DeltaIEC) mice displayed dysbiosis of the gut microbiota and increased susceptibility to DSS- and C. rodentium-induced colitis, as evidenced by disrupted epithelial antimicrobial responses, pathogen infiltration, and elevated inflammatory infiltrates in the inflamed colon. Furthermore, RNA sequencing analysis revealed significant changes in the transcriptome following GPR65 deletion in IECs, including the downregulation of immune and defense responses to bacteria. In vivo and ex vivo assays also demonstrated that GPR65 signaling was crucial for promoting antimicrobial defense, as it induced AMP production and optimized mucosal defense responses through the promotion of STAT3 phosphorylation. Additional experiments utilizing epithelial cell lines and colonoids further supported the role of GPR65 in synergizing with IL-22 to facilitate antimicrobial responses through its pH-sensing capability. Lastly, both IBD patients and DSS colitis mice exhibited decreased expression of GPR65 in inflamed epithelial cells. These findings demonstrate the important role of epithelial GPR65 in regulating intestinal homeostasis and mucosal inflammation, and suggest that targeting GPR65 may be a potential therapeutic approach in the treatment of IBD.
Pubmed: 37749885   DOI: 10.1080/19490976.2023.2257269

Mao J, et al. "GPR65 inhibits human trophoblast cell adhesion through upregulation of MYLK and ." Cell communication and signaling : CCS, 2023.
Extravillous trophoblasts (EVTs) play a crucial role in the development of the placenta by invading the maternal decidua, anchoring the placenta to the uterus, and regulating immune responses for successful implantation. The inadequate oxygen and acidic microenvironment of the decidua in early pregnancy are known to affect EVT function, but the precise mechanisms are still unclear. In this study, the impact of G protein-coupled receptor 65 (GPR65) on EVT function under acidic conditions was investigated using JAR spheroids, mouse blastocysts, and HTR-8/SVneo cells. GPR65 expression was found to be highest in EVTs and its optimal levels were essential for maintaining normal cell adhesion, migration, and invasion. Overexpression of GPR65 led to suppression of these functions by activating the cAMP-ERK signaling pathway, upregulating myosin light chain kinase (MYLK) and MYLK3 expression, and subsequently downregulating fibronectin. Consistently, elevated levels of GPR65, MYLK, and MYLK3 were observed in villus tissue samples from patients with early pregnancy loss. These findings suggest GPR65 as a potential therapeutic target for early pregnancy complications.
Pubmed: 37723567   DOI: 10.1186/s12964-023-01249-3