Abstract
Direct conversion of one cell type into another is a trans-differentiation process. Recent advances in fibroblast research revealed that epithelial cells can give rise to fibroblasts by epithelial-mesenchymal transition. Conversely, fibroblasts can also give rise to epithelia by undergoing a mesenchymal to epithelial transition. To elicit stem cell-like properties in fibroblasts, the Oct4 transcription factor acts as a master transcriptional regulator for reprogramming somatic cells. Notably, the production of gene complexes with cell-permeable peptides, such as low-molecular-weight protamine (LMWP), was proposed to induce reprogramming without cytotoxicity and genomic mutation. We designed a complex with non-cytotoxic LMWP to prevent the degradation of Oct4 and revealed that the positively charged cell-permeable LMWP helped condense the size of the Oct4-LMWP complexes (1:5 N:P ratio). When the Oct4-LMWP complex was delivered into mouse embryonic fibroblasts (MEFs), stemness-related gene expression increased while fibroblast intrinsic properties decreased. We believe that the Oct4-LMWP complex developed in this study can be used to reprogram terminally differentiated somatic cells or convert them into stem cell-like cells without risk of cell death, improving the stemness level and stability of existing direct conversion techniques.
Highlights
Accepted: 26 August 2021Trans-differentiation processes can directly convert one cell type into another [1].During these processes, transcription factors can induce the direct conversion or reprogramming of somatic cells into other differentiated lineages without an intermediate stage, which is important for the development of navigator cells [2,3,4,5]
We demonstrate that the Oct4-low-molecular-weight protamine (LMWP) complex was activated in fibroblasts, and this activation increased stem cell-like properties in this cell system
Using Fluorescence-Activated Cell Sorting (FACS), we found that 90.9% of Oct4-Lipo cells expressed CD34, while, in comparison, 74.6% of cells treated with Oct4-LMWP expressed CD34
Summary
Trans-differentiation processes can directly convert one cell type into another [1]. During these processes, transcription factors can induce the direct conversion or reprogramming of somatic cells into other differentiated lineages without an intermediate stage, which is important for the development of navigator cells [2,3,4,5]. Factordriven reprogramming of fibroblasts has been reported to directly yield other somatic cell types, such as cardiomyocytes [19], chondrocytes [20], hepatocytes [21], blood [9,22], and neural progenitors [23,24] These reports demonstrated the ability to generate progenitors and deliver stemness factors directly without establishing pluripotency. We propose a complex of the LMWP peptide and Oct gene to induce stem cell-like properties in fibroblasts. It is anticipated that the Oct4-LMWP complex can be used to reprogram terminally differentiated somatic cells or give them stem cell-like properties without risk of cell death by improving stemness level and stability of existing direct conversion technologies
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