https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/281 Sat, 04 Jul 2026 05:14:29 GMT 2026-07-04T05:14:29Z https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/212307 Title: Krüppel-like factor 4 regulates extravillous trophoblast invasion and angiogenic function and is reduced in gestational diabetes mellitus Authors: Lee, Jeonghyeon; Ryu, Ki-Young; Roh, Jaesook Abstract: IntroductionPlacental extravillous trophoblasts (EVTs) are essential for decidual invasion and spiral artery remodeling, yet the transcriptional mechanisms that sustain EVT function under metabolic stress, such as gestational diabetes mellitus (GDM), remain incompletely understood, raising the possibility that metabolically responsive transcription factors such as Krüppel-like factor 4 (KLF4) may be involved.MethodsKLF4 expression and localization were examined in human placental tissues, including early second-trimester basal plates and term placentas from normoglycemic and GDM pregnancies. Functional and molecular analyses were performed in EVT-like HTR-8/SVneo cells cultured under normal or high-glucose conditions with gain- and loss-of-function modulation of KLF4. EVT migration, invasion, and angiogenic activity were assessed using wound healing, transwell, Matrigel invasion, and tube formation assays. Transcriptional regulation was evaluated by qPCR and luciferase reporter assays.ResultsKLF4 was localized to the nuclei of EVTs in second-trimester placentas, and its expression was reduced in GDM placentas. In HTR-8/SVneo cells, high-glucose exposure (15–45 mM) suppressed KLF4 expression and reduced expression of invasion- and angiogenesis-related genes, including MMP2, MMP9, VEGFA, and PGF. Exposure to 30 mM glucose impaired EVT migration, invasion, and tube formation. Restoration of KLF4 improved migratory and angiogenic capacity. Under normal glucose conditions, KLF4 overexpression selectively increased MMP9, VEGFA, and PGF expression, whereas CRISPR–Cas9–mediated KLF4 knockdown reduced expression of all four genes. Luciferase assays demonstrated selective activation of MMP9, VEGFA, and PGF promoters by KLF4.ConclusionKLF4 supports EVT invasion and angiogenic function, and its downregulation under hyperglycemic conditions may contribute to impaired placental vascular remodeling in GDM. Fri, 01 May 2026 00:00:00 GMT https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/212307 2026-05-01T00:00:00Z https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/213287 Title: Sex-specific COX-2/CREB/ER signaling underlies male susceptibility to pulmonary fibrosis in rheumatoid arthritis-associated interstitial lung disease Authors: Kim, Misong; Kim, Kyoung-Soo; Hong, Seung-Jae; Youn, Jeehee; Kim, Young Il; Lee, Yeon-Ah Abstract: Objectives – Rheumatoid arthritis is more prevalent in women than in men, but rheumatoid arthritis-associated interstitial lung disease occurs more frequently and with greater severity in men. To address this paradox, we investigated whether sex- and tissue-specific regulation of the cyclooxygenase-2/cyclic AMP-responsive element-binding protein/estrogen receptor axis contributes to the distinct rheumatoid arthritis-associated interstitial lung disease manifestations. Methods – Curdlan-induced SKG mice were used to assess sex-dependent disease manifestations. Arthritis severity was evaluated both clinically and histologically. Pulmonary inflammation and fibrosis were analyzed histologically, whereas gene and protein expression levels were assessed by quantitative reverse transcription polymerase chain reaction and western blotting. Results – Female mice developed earlier and more severe joint arthritis, whereas male mice exhibited greater pulmonary inflammation and fibrosis. These sex- and tissue-specific differences were supported by differential expression of fibrosis-related genes (fibronectin, collagen type I alpha 1 chain, vimentin, mucin-1, and E-cadherin). Lung tissues from males showed higher estrogen receptor α and estrogen receptor β expression, while joint estrogen receptor expression was predominantly higher in females, and no sex-related differences were observed in intestinal estrogen receptor expression. Consistent with estrogen receptor expression patterns, cyclooxygenase-2 and cyclic AMP-responsive element-binding protein expression were elevated in male lungs. Collectively, these findings suggest that sex- and organ-specific activation of the cyclooxygenase-2/cyclic AMP-responsive element-binding protein/estrogen receptor axis contributes to the inflammatory patterns in Rheumatoid arthritis-associated interstitial lung disease. Conclusion – Our findings suggest that cyclooxygenase-2/cyclic AMP-responsive element-binding protein/estrogen receptor pathway activation in male lungs may explain their increased tendency toward fibrotic remodeling, whereas enhanced estrogen receptor signaling in female joints may contribute to more severe arthritis. These observations raise the possibility that sex- and tissue-specific regulation of this signaling axis influences rheumatoid arthritis-associated interstitial lung disease pathogenesis and may inform future research on therapeutic strategies. Fri, 01 May 2026 00:00:00 GMT https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/213287 2026-05-01T00:00:00Z https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/210994 Title: Caffeine enhances migration, invasion, and tube formation by upregulating MMP-2 in EVT-like HTR-8/SVneo cells Authors: Lee, Jeonghyeon; Ryu, Ki-Young; Keum, Jihyun; Roh, Jaesook Abstract: Caffeine is consumed widely during pregnancy, yet its effects on cellular processes relevant to early placental development, particularly on extravillous trophoblast (EVT) function, remain incompletely understood. This study examined the impact of caffeine on EVT-like HTR-8/SVneo cells, focusing on key cellular processes including migration, invasion, and matrix-interactive behaviors relevant to trophoblast function. Treatment with 1 mM caffeine enhanced cell motility and tube formation in EVT-like cells without affecting cell viability or inducing apoptosis. These functional changes were associated with a selective upregulation of MMP-2 expression and activity, while MMP-9 remained unchanged. Pharmacological inhibition of MMP-2 abolished caffeine-induced increases in migration, invasion, and tube formation, supporting a functional role for MMP-2 in these responses. At the signaling level, caffeine-induced MMP-2 upregulation was attenuated by inhibition of PI3K/Akt signaling, with a minor contribution from PKA, whereas MEK1/2 inhibition had minimal effect, supporting the involvement of distinct signaling pathway in EVT-like cells. The observed trophoblast-specific response contrasts with the inhibitory effects of caffeine reported in cancer and endothelial cells. Collectively, these findings identify a previously uncharacterized mechanism by which caffeine modulates signaling and matrix-remodeling−associated behaviors in EVT-like cells in vitro. Sun, 01 Mar 2026 00:00:00 GMT https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/210994 2026-03-01T00:00:00Z https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/211375 Title: Caffeine Exposure Modulates Trophoblast Differentiation and Estradiol Synthesis Authors: Keum, Jihyun; Lee, Jeonghyeon; Ryu, Ki-Young; Roh, Jaesook Abstract: Differentiation of villous cytotrophoblasts into syncytiotrophoblasts is essential for placental endocrine function and estradiol production. Caffeine consumption has been linked to altered estradiol levels, but its effects on human trophoblast differentiation remain incompletely understood. This study investigated the effects of caffeine on biochemical differentiation of human trophoblasts using BeWo cells and human placental explants. Cell viability and apoptosis were assessed using CCK-8 and in situ TUNEL assays. Differentiation-associated changes were evaluated by measuring CYP19A1 and its placenta-specific promoter transcript CYP19 I.1, at the mRNA level, while aromatase protein expression and estradiol production were assessed by Western blotting and ELISA, respectively. Exposure to 2 mM caffeine reduced BeWo cell viability, whereas 1 mM caffeine had no detectable effects on cell viability or apoptosis. At non-cytotoxic concentrations, caffeine significantly increased CYP19A1 mRNA expression under both basal and forskolin-stimulated conditions and elevated estradiol production. Similar transcriptional and endocrine responses were observed in human placental explants. Pharmacological inhibition demonstrated that caffeine-induced CYP19A1 transcriptional upregulation was dependent on PKA signaling, but not on PKC or MAPK pathways. These results indicate that caffeine can modulate trophoblast biochemical differentiation via PKA-dependent regulation of placental aromatase expression and estradiol synthesis. While these findings provide mechanistic insight into caffeine-mediated effects on trophoblast endocrine function, their relevance to physiological exposure levels and in vivo placental development warrants cautious interpretation. Sun, 01 Feb 2026 00:00:00 GMT https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/211375 2026-02-01T00:00:00Z