Combining RNAscope, Immunohistochemistry (IHC) and Digital Image Analysis to Assess Podoplanin (PDPN) Protein and PDPN_mRNA Expression on Formalin-Fixed Paraffin-Embedded Normal Human Placenta Tissues

• Published in: Current issues in molecular biology Vol. 46; no. 6; pp. 5161 - 5177
• Main Authors: Tomescu, Larisa Cristina, Cosma, Andrei Alexandru, Fenesan, Mihaela Pasca, Melnic, Eugen, Petrovici, Vergil, Sarb, Simona, Chis, Monica, Sas, Ioan, Ribatti, Domenico, Cimpean, Anca Maria, Dorobantu, Florica Ramona
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI 24.05.2024; MDPI AG
• Subjects: decidua; normal human placenta; placental villi; podolanin; decidua; normal human placenta; placental villi; podolanin
• ISSN: 1467-3045; 1467-3037; 1467-3045
• DOI: 10.3390/cimb46060310

The expression and function of podoplanin (PDPN) in the normal human placenta has been debated in placental evaluation. This study emphasizes the importance of a multimodal approach of PDPN expression in normal human placentas. A complete examination is performed using immunohistochemistry, RNAscope and automated Digital Image examination (DIA) interpretation. QuPath DIA-based analysis automatically generated the stromal and histological scores of PDPN expression for immunohistochemistry and RNAscope stains. The umbilical cord's isolated fibroblasts and luminal structures expressed PDPN protein and PDPN_mRNA. RNAscope detected PDPN_mRNA upregulation in syncytial placental knots trophoblastic cells, but immunohistochemistry did not certify this at the protein level. The study found a significant correlation between the IHC and RNAscope H-Score ( = 0.033) and Allred Score ( = 0.05). A successful multimodal strategy for PDPN assessment in human placentas confirmed PDPN expression heterogeneity in the full-term human normal placenta and umbilical cord at the protein and mRNA level. In placental syncytial knots trophoblastic cells, PDPN showed mRNA overexpression, suggesting a potential role in placenta maturation.