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Transcriptomic effects of intermittent parathyroid hormone treatment on Sox9-lineage osteoprogenitors

ABSTRACT: We employed 10X Genomics single cell RNA-sequencing to study the bone anabolic effects of intermittent parathyroid hormone treatment (iPTH) on growth-associated progenitors. These osteoblast progenitors, marked by the expression of the transcription factor SRY-box transcription factor 9 (Sox9), reside in the growth plate and perichondrium that respond to iPTH in early postnatal mice. Whether these cells respond in adult mice is unknown. Using lineage tracing approach with Sox9-CreERT2 crossed with Ai9 tdTomato reporter mice, we traced Sox9-expressing cells and their descendants following 10 days of vehicle or iPTH treatment. This study represents a single cell atlas of Sox9-lineage cells following 10 days of iPTH treatment.

ORGANISM(S): Mus musculus

PROVIDER: GSE326229 | GEO | 2026/09/20

REPOSITORIES: GEO

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			Action	DRS
		Other
	GSE326229_adult_sox9_barcodes.tsv.gz	Tabular
	GSE326229_adult_sox9_cell_metadata.csv.gz	Csv
	GSE326229_adult_sox9_genes.tsv.gz	Tabular
	GSE326229_adult_sox9_matrix.mtx.gz	Other

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Project description:Primary hyperparathyroidism is a common endocrine disorder frequently affecting postmenopausal women. In this study we have investigated expression of the prolactin receptor (PRLr) in a panel of 37 sporadic parathyroid tumours, as well as functionality in vitro in cultured parathyroid tumour cells. High levels of the prolactin receptor gene (PRLR) transcripts were demonstrated in parathyroid tissues as compared to other reference tissues and breast cancer cells. PRLr products of 60/70 kDa were highly expressed in all parathyroid tumours. In addition varying levels of the 80 kDa PRLr isoform, with known proliferative activity, were demonstrated. In parathyroid tumours PRLr immunoreactivity was observed in cytoplasm in all cases and in addition in the plasma membrane (n = 12) or enlarged lysosomes (n = 4). In normal parathyroid rim PRLr was expressed in cytoplasm and granulae. In in vitro studies of short-term cultured human parathyroid tumour cells prolactin stimulation was associated with transcriptional changes in JAK/STAT, RIG-I like receptor and type II interferon signaling pathways as documented by gene expression profiling. Moreover, PRLR gene expression in parathyroid tumors was significantly inversely correlated with plasma total Ca2+ levels. In conclusion, the prolactin receptor was found highly abundant in human parathyroid gland, parathyroid tumours, correlated with patient Ca2+ levels and functionally responsive to physiological levels of prolactin. These findings suggest a role for the prolactin receptor in human parathyroid adenomas. Expression profiling was done in parathyroid adenomas subjected to prolactin treatment in culture. In addition, corresponding paraffin sections were obtained for verification of PRLr expression by immunohistochemistry. 200 mg/L prolactin (recombinant humanM-BM- prolactin, Cat. No. JM-4687-50, MBL Woburn, MA) was added to 1M-CM-^W 10^6 attached parathyroid tumour cells. Cells were harvested using RNAlater (QIAGEN) and homogenized with QIAshredder for RNA extraction after 3 h and 24 h in culture, respectively. Negative controls were collected in parallel with each case at the same time points. RNA was extracted using QIA Cube, and quality assessed with Bioanalyser and Nanodrop. Expression array profiling and data analysis was done at the KI core facility Bioinformatics and Expression Analysis (BEA, Novum, Huddinge) using the Affymetrix platform and the TITAN ST 1.1 array. A total of 16 samples were analysed including 4 parathyroid adenomas cultured for 3 h or 24 h in the presence of prolactin plus control samples cultured in parallel without prolactin.

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Transcriptomic effects of intermittent parathyroid hormone treatment on Sox9-lineage osteoprogenitors

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