Kanaga Arul Nambi Rajan1, Marwa Khater2, Francesca Soncin1, Donald Pizzo3, Matteo Moretto-Zita1, Jonathan Pham1, Orysya Stus1, Pooja Iyer3, Veronique Tache4, Louise C Laurent2, Mana M Parast5. 1. Department of Pathology, University of California San Diego, La Jolla, CA, 92093, USA; Sanford Consortium for Regenerative Medicine, University of California San Diego, La Jolla, CA, 92093, USA. 2. Sanford Consortium for Regenerative Medicine, University of California San Diego, La Jolla, CA, 92093, USA; Department of Reproductive Medicine, University of California San Diego, La Jolla, CA, 92093, USA. 3. Department of Pathology, University of California San Diego, La Jolla, CA, 92093, USA. 4. Department of Obstetrics and Gynecology, University of California Davis, Sacramento, CA, 95817, USA. 5. Department of Pathology, University of California San Diego, La Jolla, CA, 92093, USA; Sanford Consortium for Regenerative Medicine, University of California San Diego, La Jolla, CA, 92093, USA. Electronic address: mparast@ucsd.edu.
Abstract
INTRODUCTION: Placental insufficiency, arising from abnormal trophoblast differentiation and function, is a major cause of fetal growth restriction. Sirtuin-1 (Sirt1) is a ubiquitously-expressed NAD-dependent protein deacetylase which plays a key role in numerous cellular processes, including cellular differentiation and metabolism. Though Sirt1 has been widely studied, its role in placentation and trophoblast differentiation is unclear. METHOD: Sirt1-heterozygous mice were mated and evaluated at various points during embryogenesis. In situ hybridization and immunohistochemistry were used to further characterize the placental phenotype of Sirt1-null mice. Wild-type (WT) and Sirt1-null mouse trophoblast stem cell (TSC) lines were derived from e3.5 littermate blastocysts. These cells were then evaluated at various points following differentiation. Differentiation was evaluated by expression of lineage specific markers using qPCR and flow cytometry, as well as Matrigel invasion assays. Global gene expression changes were evaluated using microarray-based RNA profiling; changes in specific pathways were validated using qPCR and western blot. RESULTS: In the absence of Sirt1, both embryos and placentas were small, with placentas showing abnormalities in both the labyrinthine layer and junctional zone. Sirt1-null TSCs exhibited an altered phenotype in both undifferentiated and differentiated states, phenotypes which corresponded to changes in pathways relevant to both TSC maintenance and differentiation. Specifically, Sirt1-null TSC showed blunted differentiation, and appeared to be suspended in an Epcamhigh trophoblast progenitor state. DISCUSSION: Our results suggest that Sirt1 is required for proper TSC differentiation and placental development.
INTRODUCTION: Placental insufficiency, arising from abnormal trophoblast differentiation and function, is a major cause of fetal growth restriction. Sirtuin-1 (Sirt1) is a ubiquitously-expressed NAD-dependent protein deacetylase which plays a key role in numerous cellular processes, including cellular differentiation and metabolism. Though Sirt1 has been widely studied, its role in placentation and trophoblast differentiation is unclear. METHOD:Sirt1-heterozygous mice were mated and evaluated at various points during embryogenesis. In situ hybridization and immunohistochemistry were used to further characterize the placental phenotype of Sirt1-null mice. Wild-type (WT) and Sirt1-null mouse trophoblast stem cell (TSC) lines were derived from e3.5 littermate blastocysts. These cells were then evaluated at various points following differentiation. Differentiation was evaluated by expression of lineage specific markers using qPCR and flow cytometry, as well as Matrigel invasion assays. Global gene expression changes were evaluated using microarray-based RNA profiling; changes in specific pathways were validated using qPCR and western blot. RESULTS: In the absence of Sirt1, both embryos and placentas were small, with placentas showing abnormalities in both the labyrinthine layer and junctional zone. Sirt1-null TSCs exhibited an altered phenotype in both undifferentiated and differentiated states, phenotypes which corresponded to changes in pathways relevant to both TSC maintenance and differentiation. Specifically, Sirt1-null TSC showed blunted differentiation, and appeared to be suspended in an Epcamhigh trophoblast progenitor state. DISCUSSION: Our results suggest that Sirt1 is required for proper TSC differentiation and placental development.
Authors: Francesca Soncin; Marwa Khater; Cuong To; Donald Pizzo; Omar Farah; Anna Wakeland; Kanaga Arul Nambi Rajan; Katharine K Nelson; Ching-Wen Chang; Matteo Moretto-Zita; David R Natale; Louise C Laurent; Mana M Parast Journal: Development Date: 2018-01-29 Impact factor: 6.868
Authors: Abdelrahim Alqudah; Kelly-Ann Eastwood; Djurdja Jerotic; Naomi Todd; Denise Hoch; Ross McNally; Danilo Obradovic; Stefan Dugalic; Alyson J Hunter; Valerie A Holmes; David R McCance; Ian S Young; Chris J Watson; Tracy Robson; Gernot Desoye; David J Grieve; Lana McClements Journal: Front Endocrinol (Lausanne) Date: 2021-06-02 Impact factor: 5.555