TP63 basal cells are indispensable during endoderm differentiation into proximal airway cells on acellular lung scaffolds

Abstract

The use of decellularized whole-organ scaffolds for bioengineering of organs is a promising avenue to circumvent the shortage of donor organs for transplantation. However, recellularization of acellular scaffolds from multicellular organs like the lung with a variety of different cell types remains a challenge. Multipotent cells could be an ideal cell source for recellularization. Here we investigated the hierarchical differentiation process of multipotent ES-derived endoderm cells into proximal airway epithelial cells on acellular lung scaffolds. The first cells to emerge on the scaffolds were TP63+ cells, followed by TP63+/KRT5+ basal cells, and finally multi-ciliated and secretory airway epithelial cells. TP63+/KRT5+ basal cells on the scaffolds simultaneously expressed KRT14, like basal cells involved in airway repair after injury. Removal of TP63 by CRISPR/Cas9 in the ES cells halted basal and airway cell differentiation on the scaffolds. These findings suggest that differentiation of ES-derived endoderm cells into airway cells on decellularized lung scaffolds proceeds via TP63+ basal cell progenitors and tracks a regenerative repair pathway. Understanding the process of differentiation is key for choosing the cell source for repopulation of a decellularized organ scaffold. Our data support the use of airway basal cells for repopulating the airway side of an acellular lung scaffold.

Integrated Genomics Approaches Identify Transcriptional Mediators and Epigenetic Responses to Afghan Desert Particulate Matter in Small Airway Epithelial Cells

“…P53/P63 signaling responses to APM were identified by interrogation of dynamic TREs from PRO-seq and chromatin accessibility features from ATAC-seq, suggesting a combined signal-dependent and lineage-determining role for P53/P63 in small airway epithelia progenitor cells. This latter function aligns with numerous previous studies of the airway epithelia (43, 44, 46, 48), however, the integrated role of this family in exposure responses has not been reported to our knowledge. The combined role may reflect differential activity of distinct P53/P63 family members or the effects of transcription factors that cooperate with P53/P63.…”

Decellularization of porcine kidney with submicellar concentrations of SDS results in the retention of ECM proteins required for the adhesion and maintenance of human adult renal epithelial cells

“…This lack of organization is in contrast to our previous studies with kidney and lung, where recellularization using pluripotent cell-derived organ progenitor cells were used. 22,41,73 In those studies, we observed that kidney progenitor cells derived from embryonic stem cells could adhere to the ECM at any location and would then be directed to differentiate towards a specific kidney cell type specified by the ECM proteins unique to that location within the kidney, such as a proximal tubule or glomerulus ECM site. 41 Here, we demonstrate that adult human renal epithelial cells are compatible with porcine ECM, but are unable to organize on the ECM, which has both in vitro and clinical implications when considering cell types for recellularization in the generation of de novo tissue and organs.…”

Molecular Mechanisms and Cellular Contribution from Lung Fibrosis to Lung Cancer Development

“…This molecular and omics approach allowed the identification of the basaloid cells, a novel and rare cell population detected in the lungs of IPF patients. These cells express some of the common airway basal cell markers (TP63, KRT17, and LAMB3 LAMC2) and SOX9 transcription factor, crucial for airway development and repair, but lack some lung keratins, such as KRT5 and KRT15 [259]. Thus, basaloid cells could represent a novel source of cells to direct the efforts for future target therapies.…”