Spatial profiling of scRNAseq signatures in human parathyroid glands
Awardee: Julie Ann Sosa
Institution: University of California at San Francisco
Grant Amount: $50,000
Funding Period: April 1, 2021 - March 31, 2022
The objectives of this project were to: (1) utilize transcriptomic methods to define individual cell types within the human parathyroid, and (2) employ digital spatial profiling to visualize the localization of these cell types within the native parathyroid gland architecture. The developmental pilot phase work supported by the grant enabled us to establish a solid foundation of procedural optimization and proof of concept data for scaling our single cell sequencing efforts to a larger, more broadly representative cohort of donor parathyroid glands.
The scientific objectives completed during the one-year project period are essential for comprehensive mapping of the human parathyroid gland. The specific landmarks achieved include: demonstration that our live organ procurement work flow preserves tissue viability and maintains intact biochemical function; validation of recovery efficiency, parathyroid marker expression and cellular integrity in suspension; comparative assessment of whole cell vs nuclear isolation for downstream molecular analysis; validation of a novel split-pool sequencing approach that greatly improves capture efficiency, reduces selective recovery bias, and eliminates library construction batch effect concerns; digital spatial profiling of archived normal parathyroid gland sections to demonstrate the capture and whole transcriptome interrogation of specific cellular subsets demarcated by marker gene expression; and the molecular data from these studies showing that the cellular composition and transcriptional profiles of parathyroid gland tissue are dynamic rather than static. This last finding reveals that the cellular content and biochemical activity of the parathyroid gland may be physiologically conditional, suggesting that functional reconstitution of the parathyroid gland is not a fixed target, but instead requires complementation of adaptive capacity in addition to terminally differentiated cellular phenotypes. These key data will inform future and ongoing studies to reconstitute native parathyroid gland function.
Publication:
Chia-Ling Tu, Wenhan Chang, Julie A Sosa, James Koh
PNAS Nexus, Volume 2, Issue 3, March 2023, pgad073