Radio Electric Conveyed Fields Directly Reprogram Human Dermal Skin Fibroblasts toward Cardiac, Neuronal, and Skeletal Muscle-Like Lineages

Abstract

Somatic cells can be directly reprogrammed to alternative differentiated fates without first becoming stem/progenitor cells. Nevertheless, the initial need for viral-mediated gene delivery renders this strategy unsafe in humans. Here, we provide evidence that exposure of human skin fibroblasts to a Radio Electric Asymmetric Conveyer (REAC), an innovative device delivering radio electric conveyed fields at a radiofrequency of 2.4 GHz, afforded remarkable commitment toward cardiac, neuronal, and skeletal muscle lineages. REAC induced the transcription of tissue-restricted genes, including Mef2c, Tbx5, GATA4, Nkx2.5, and prodynorphin for cardiac reprogramming, as well as myoD, and neurogenin 1 for skeletal myogenesis and neurogenesis, respectively. Conversely, REAC treatment elicited a biphasic effect on a number of stemness-related genes, leading to early transcriptional increase of Oct4, Sox2, cMyc, Nanog, and Klf4 within 6-20 h, followed by a downregulation at later times. The REAC action bypassed a persistent reprogramming toward an induced pluripotent stem cell-like state and involved the transcriptional induction of the NADPH oxidase subunit Nox4. Our results show for the first time the feasibility of using a physical stimulus to afford the expression of pluripotentiality in human adult somatic cells up to the attainment of three major target lineages for regenerative medicine.

Stem cell senescence. Effects of REAC technology on telomerase-independent and telomerase-dependent pathways

“…These results are consistent with our previous observations showing that REAC TO-RGN exposure modulated the expression of Nanog, Sox2, Oct4, and cMyc in mouse embryonic stem cells, as well as in human skin-derived fibroblasts3132. Furthermore, REAC TO-RGN exposure proved effective in antagonizing the aging process in vitro 333435.…”

“…Moreover, exposure to REAC afforded a direct reprogramming of human dermal skin fibroblasts32 ensuing into a rapid, high-throughput and stable commitment towards cardiogenesis, skeletal myogenesis, and neurogenesis32. REAC protocols of regenerative medicine have also shown a clear efficacy in abating the effects of the aging process in vitro 333435.…”

Cancer Stem Cells: Foe or Reprogrammable Cells for Efficient Cancer Therapy?

“…Exposure to ACEF was able to induce a biphasic effect, i.e. overexpression followed by transcriptional inhibition of Sox2, Nanog, Oct3/4, Klf4 and c-myc in human skin fibroblasts, affording for the first time a direct high-yield reprogramming of adult nonstem somatic cells into myocardial, neural and skeletal muscle cells (about 15-20% for each phenotypic commitment) [76].…”

Seeing Cell Biology with the Eyes of Physics

“…Commitment towards the same lineages has been achieved by REAC treatment of human skin fibroblasts [38]. For the first time, human non-stem somatic adult cells were directed to fates into which they would never otherwise be committed to, avoiding practices that cannot so far be readily envisioned in a clinical setting, such as the gene delivery by lentiviral vectors, or the cellular reprogramming by cumbersome and expensive chemistry, including the use of non-integrating technologies.…”

“…For the first time, human non-stem somatic adult cells were directed to fates into which they would never otherwise be committed to, avoiding practices that cannot so far be readily envisioned in a clinical setting, such as the gene delivery by lentiviral vectors, or the cellular reprogramming by cumbersome and expensive chemistry, including the use of non-integrating technologies. Moreover, somatic cell reprogramming by the REAC technology was mediated by a transient overexpression of pluripotency genes, followed by their down-regulation [38], without halting the treated cells into an embryonic-like state which may drift into cancer or unwanted cell types.…”