Identification of Early Transcriptional Markers of ADPKD Cystic Epithelial Cells
- Genetic Diseases: Cystic - Basic
November 03, 2023 | Location: Exhibit Hall, Pennsylvania Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Genetic Diseases of the Kidneys
- 1201 Genetic Diseases of the Kidneys: Cystic
- Rai, Victoria, Yale University, New Haven, Connecticut, United States
- Onuchic, Laura, Yale University, New Haven, Connecticut, United States
- Reyna-Neyra, Marcela Andrea, Yale University, New Haven, Connecticut, United States
- Schena, Giorgia, Yale University, New Haven, Connecticut, United States
- Craft, Joseph E., Yale University, New Haven, Connecticut, United States
- Caplan, Michael J., Yale University, New Haven, Connecticut, United States
Mechanisms driving ADPKD cystogenesis are not well understood, and reliable biomarkers are scarce. To explore molecular changes in cystogenesis, we performed single cell RNA sequencing on kidneys from an ADPKD mouse model at an early stage of cyst formation and from a model in which cystic disease was suppressed by expression of the polycystin-1 C-terminal tail (PC1-CTT).
Doxycycline inducible Pkd1fl/fl;Pax8rtTA;Teto-Cre ADPKD C57BL/6N mice expressing CTT (Pkd1-KO+CTT) or not (Pkd1-KO) were induced between 4-6 weeks and aged to 10 weeks. Single cell kidney suspensions from biological replicates were pooled and processed for cDNA synthesis and library preparation with 10X Chromium technology. Reads were aligned using the 10X Genomics CellRanger pipeline and further processed using Seurat v4.3.
Clusters corresponding to tubule segments were identified using established annotations. Several novel clusters defined by expression of multiple markers, including Jarid2, Tmtc2, and Filip1l, were identified in the Pkd1-KO model. These clusters were sparsely populated in Pkd1-KO+CTT samples, suggesting that they may correspond to cyst epithelial cells that are suppressed by PC1-CTT expression. Each of these novel clusters included cells whose markers identified them from distinct tubule segments (e.g. Sglt2, Umod, S100g), suggesting that cells of different segments are clustered together by virtue of sharing the putative cyst cell transcriptional signature. The single cluster observed in Pkd1-KO+CTT model and not in the Pkd1-KO model is enriched in markers associated with metabolic processes including Ugt1a2, Cbr1, and Pck1. Importantly, at 10 weeks, there are no significant differences in %KW/BW or kidney morphology between the Pkd1-KO+CTT and Pkd1-KO models, suggesting that the cells belonging to the cystic epithelial cell clusters exhibit early identifiable differences in transcriptional programming.
scRNA-seq at an early timepoint reveals novel transcriptional makers of putative cystic epithelial cells. The protein expression levels of individual markers are being assessed. These results suggest that there are epithelial cells early in disease progression exhibiting a unique transcriptional signature that may elucidate the mechanisms of cyst formation and that may provide new candidate biomarkers.
- NIDDK Support