Proteomic profiling of key transcription factors in the process of neonatal mouse cardiac regeneration capacity loss

Abstract

The heart lacks complete regeneration capacity. In mice, the cardiac apex can regenerate 1 day after birth, although 7 days after birth the repair occurs with a fibrous scar. However, the key transcription factors (TFs) related to this loss of regeneration capacity remain largely unknown. We aimed to find candidates for key TFs using proteomic profiling and comparison during loss of neonatal mouse cardiac regeneration capacity, with preliminary validation using western blotting (WB) and real-time quantitative polymerase chain reaction (RT-qPCR). A total of 69 common discrepant TFs with similar variation trends in two TF response element experiments were identified, 18 of which were matched to known key signaling pathways of cardiac regeneration after pathway enrichment of downstream genes. Validation using RT-qPCR-selected DACH1, RBL1 (P107), and TBX20, and further validation with WB-selected RBL1 (P107) and TBX20. We therefore identified two candidates for key TFs in the loss of mouse cardiac apex regeneration capacity. TBX20 has been biologically validated, and RBL1 (P107) needs to be validated in the future.Abbreviations: catTFREs, concatenated tandem array of 50 consensus TFREs; FOT, frequency of total; LC-MS, nano-liquid chromatography/tandem mass spectrometry; RT-qPCR, real-time quantitative polymerase chain reaction; TFRE, TF response elements; TFs, transcription factors; WB, western blotting First author under standardized training of residents at West China hospital.

Cell-Type-Specific Gene Regulatory Networks Underlying Murine Neonatal Heart Regeneration at Single-Cell Resolution

“…Although these studies have shed light on potential regulators of regeneration, a more comprehensive and system-level view is needed to understand the intrinsic differences between regenerative and non-regenerative hearts and their differential responses to injury. To address this issue, genomic and proteomic approaches have been applied to profile changes in gene expression, epigenetic landscapes, and protein abundance during neonatal heart regeneration using bulk tissues or flow-cytometry-sorted major cell types ( Fan et al, 2020 ; Quaife-Ryan et al, 2017 ; Wang et al, 2019a , 2019b ). These studies generated valuable insights into the distinctive neonatal features associated with heart regeneration at the systems biology level but because of technical limitations were unable to resolve the individual contributions of different cardiac cell subtypes to the underlying regenerative processes.…”

Proteome-scale profiling reveals MAFF and MAFG as two novel key transcription factors involved in palmitic acid-induced umbilical vein endothelial cell apoptosis

“…The catTFRE-bead complex was then mixed with the nuclear protein extracts and incubated at 4 °C for 2 h to enrich the transcription factors in the nuclear protein extracts. The detailed TFRE procedures are described in our previous study [7].…”