Panoply™ Human MANF Over-expressing Stable Cell Line

Abnormal development and defective invasion of early trophoblast cells are associated with unexplained recurrent miscarriage (URM). Although mesencephalic astrocyte-derived neurotrophic factor (MANF) is known to inhibit the migration and invasion of certain cancer cells, its role in pregnancy-related disorders remains unclear. Here, researchers found that MANF levels in the peripheral blood and miscarriage tissues of women with URM were significantly elevated compared to normal controls. They demonstrated that MANF interacts with nucleophosmin 1 (NPM1) in the trophoblast cells of URM patients; this interaction promotes the ubiquitin-mediated degradation of NPM1, thereby upregulating the p53 signaling pathway and inhibiting cellular proliferation, migration, and invasion. Using a mouse model of URM, the study revealed that downregulating MANF reduced fetal resorption (i.e., lowered the risk of miscarriage); however, the simultaneous downregulation of NPM1 led to an increased miscarriage rate. These data indicate that MANF induces miscarriage by downregulating NPM1 and activating the p53 pathway. Consequently, downregulating MANF or blocking the MANF-NPM1 interaction represents a potential therapeutic strategy for URM.

Here, the researchers generated MANF-overexpressing or MANF-knockdown HTR-8/SVneo cell lines (Figure 1A) and examined MMP2 and MMP9 expression at varying MANF levels. The results showed that MMP2 and MMP9 expression decreased as MANF levels increased; conversely, levels of these MMPs rose when MANF was knocked down. Furthermore, using EdU and TUNEL staining to assess proliferation and apoptosis, the researchers found that MANF overexpression inhibited cell proliferation and promoted apoptosis (Figure 1B and C). Wound healing and Transwell assays demonstrated that MANF overexpression impaired cell migration and invasion, whereas MANF knockdown enhanced these capabilities (Figure 1D and E).

Figure 1. MANF expression levels differentially regulate MMP expression, as well as cell invasion and migration abilities. (Fang Y, et al., 2024)