Abstract: SA-PO543

Understanding Normal and Hoxa9,10,11-/-; Hoxd9,10,11-/- Kidney Development Through Single Cell RNA-Seq

Session Information

  • Developmental Biology
    November 04, 2017 | Location: Hall H, Morial Convention Center
    Abstract Time: 10:00 AM - 10:00 AM

Category: Developmental Biology and Inherited Kidney Diseases

  • 401 Developmental Biology

Authors

  • Magella, Bliss, Cincinnati Children''s Hospital Medical Center, Sharonville, Ohio, United States
  • Mahoney, Robert, Cincinnati Childrens, Fairfield, Ohio, United States
  • Venkatasubramanian, Meenakshi, Cincinnati Children''s Hospital, Cincinnati, Ohio, United States
  • Salomonis, Nathan, Cincinnati Children''s Hospital Medical Research Center, Cincinnati, Ohio, United States
  • Potter, Steven, Cincinnati Children's Hospital, Cincinnati, Ohio, United States
Background

The kidney is a complex organ that is made of many different cell types. In an effort to better understand the cell diversity within the developing kidney we have performed single cell RNA-seq on embryonic day 14.5 mouse kidneys. Of particular interest is the apparent lack of a Hox code within the developing kidney. Previous studies have determined that Hox 10 and 11 paralogous groups are functioning within kidney development. Interestingly, thirty-six of the thirty-nine mammalian Hox genes are expressed during kidney development.

Methods

Analysis of wild type and mutant E14.5 kidneys was performed using data obtained through Drop-seq. Additional data sets were gathered using Fluidigm and Chromium In-Drop for wild type validation. Hoxa9,10,11; Hoxd9,10,11 mutants were used to determine the functional role of the Hox genes during kidney development. Altanalyze and Seurat based analyses are being used to determine the normal and mutant expression profiles.

Results

Sixteen cell groups were identified in the wild type data set; medullary collecting duct, cortical collecting duct, ureteric bud tip, loop of Henle, distal comma shaped body, podocyte, mid S-shaped body, early proximal tubule, pre-tubular aggregate, three cap mesenchyme groups, endothelium, nephrogenic zone stroma, cortical stroma, and medullary stroma. In addition to the known identifier genes, novel specific gene associations were also discovered during analysis. One such example is the discovery of Gdnf expression from within the stromal population, which was previously thought of as being exclusively expressed from the cap mesenchyme. Morphological analysis of the Hox mutants shows alterations in mature nephron segment identity, medullary zone specification, and the formation of a pelvic opening.

Conclusion

Obtaining a comprehensive data set allows for the visualization of the expression profile of many genes within the wild type developing kidney. These data can then be used as an atlas for comparing mutant data sets. By combining the Hox mutant morphological phenotypes with the single cell expression profiles we can better understand the molecular regulation of kidney development. This greater understanding provides necessary information for future studies looking to make kidneys through organoid cultures.

Funding

  • NIDDK Support