8649 Ablation of Leptin Receptors in Somatotropes Impacts Transcriptomic Plasticity in the Pou1f1 Lineage of Female Pituitary Cells
Authors: Miles Tiffany K , Odle Angela Katherine , Byrum Stephanie , Lagasse Alex , Haney Anessa , Ortega Victoria G , Herdman Ashley , MacNicol Melanie C , MacNicol Angus M , Childs Gwen V 
Published: October 2024

Disclosure: T.K. Miles: None. A.K. Odle: None. S. Byrum: None. A. Lagasse: None. A. Haney: None. V.G. Ortega: None. A. Herdman: None. M.C. MacNicol: None. A.M. MacNicol: None. G.V. Childs: None.

Anterior pituitary somatotropes produce growth hormone (GH) to optimize body composition, receiving tropic signals from leptin about the metabolic state. When leptin receptors are ablated in somatotropes, GH proteins, serum GH and GHRH receptors are reduced. Unexpectedly, POU1F1, Prolactin, and TSH-beta proteins are also reduced in pituitaries of Somatotrope LEPR-null females (mutants), suggesting a tropic role for leptin in differentiation. To test this hypothesis, single cell RNA-seq analysis compared the pituitary transcriptome of 6-month-old control and mutant female mice housed under thermoneutral conditions (28°C). Following scRNA-seq, cells were clustered computationally with the use of Seurat; all cell types were identified in separate UMAP clusters. Mutants showed increases in cell numbers in somatotrope (4.7X) and Pou1f1 (5.7x) clusters, but no change in thyrotrope or lactotrope clusters. In LEPR-null female somatotropes, the 516 differentially expressed genes (DEGs; adj p&lt;0.05) included upregulated Pomc, Prl, Lhb, Tshb and Cga (Log2FC+0.5-1.7) and downregulated Pou1f1, Ghrhr, and Gh (Log2FC -0.5—1.5). In the lactotrope cluster from mutants, Pomc, Cga, and Gh were upregulated (Log2FC+1.1-1.5) and Pou1f1 and Prl were downregulated. The thyrotrope cluster was unchanged. Prl, Pomc, and Gh were upregulated in the Pou1f1 cluster. The down regulation of Gh, Pou1f1, and Ghrhr in somatotropes and Prl in lactotropes correlates with the lower serum levels of GH and Prl in mutant females. However, the upregulated DEGs do not correlate with increased secretory activity suggesting that the multihormonal cells are not fully functional. This was confirmed with Ingenuity Pathway Analysis (IPA) showing downregulation of canonical pathways involved in secretion in mutant somatotropes. Unexpectedly, mutant females exposed to a HFD (60% fat) for 16 weeks (+16.6g) showed a reversal of the changes in somatotrope cell numbers and multihormonal expression seen in control fed mutants. The increase in genes belonging to IPA canonical cytoplasmic translation pathways in mutant somatotropes were also completely reversed following a HFD. Furthermore, the downregulated secretory/signaling pathways seen in control fed mutant female somatotropes and lactotropes (endocytosis, vesicle transport, Gaq, GnRH) were upregulated modestly following a HFD. The high serum leptin in the HFD fed mutant mice may activate signaling pathways in cells still expressing LEPR. Collectively, these results confirm a role for leptin in maintaining differentiated somatotropes. They also suggest that leptin may limit expansion of multihormonal progenitor cells. The decreased multihormonal expression and protein synthesis in lactotrope clusters from mutant mice exposed to a HFD are also seen in lactotropes from control mice on a HFD, which suggests that a HFD may compromise pituitary plasticity.

Presentation: 6/3/2024