Journal
KIDNEY INTERNATIONAL
Volume 95, Issue 5, Pages 1153-1166Publisher
ELSEVIER SCIENCE INC
DOI: 10.1016/j.kint.2018.11.041
Keywords
cap mesenchyme; direct reprogramming; nephron progenitor; reprogramming; transposon
Categories
Funding
- Australian Cancer Research Foundation
- Victorian Government's Operational Infrastructure Support Program
- National Health and Medical Research Council of Australia [GNT1041275]
- National Institutes of Health [2T32DK060445-11, DK093660]
- Vanderbilt O'Brien Renal Center [P30 DK114809]
- Department of Veterans Affairs [BX002797, BX002190, BX004258]
- Vanderbilt Center for Kidney Disease
- NHMRC [APP1042093, APP1136085]
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All nephrons in the mammalian kidney arise from a transient nephron progenitor population that is lost dose to the time of birth. The generation of new nephron progenitors and their maintenance in culture are central to the success of kidney regenerative strategies. Using a lentiviral screening approach, we previously generated a human induced nephron progenitor-like state in vitro using a pool of six transcription factors. Here, we sought to develop a more efficient approach for direct reprogramming of human cells that could be applied in vivo. PiggyBac transposons are a non-viral integrating gene delivery system that is suitable for in vivo use and allows for simultaneous delivery of multiple genes. Using an inducible piggyBac transposon system, we optimized a protocol for the direct reprogramming of HK2 cells to induced nephron progenitor-like cells with expression of only 3 transcription factors (SNAI2, EYA1, and SIX1). Culture in conditions supportive of the nephron progenitor state further increased the expression of nephron progenitor genes. The refined protocol was then applied to primary human renal epithelial cells, which integrated into developing nephron structures in vitro and in vivo. Such inducible reprogramming to nephron progenitor-like cells could facilitate direct cellular reprogramming for kidney regeneration.
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