Abstract
Background: Cardiovascular disease remains the leading cause of global mortality, with myocardial fibrosis representing a critical barrier to cardiac regeneration. Human cardiac fibroblasts constitute the predominant non-myocyte population in the heart and are principal mediators of pathological fibrosis following injury. Their direct reprogramming toward cardiac progenitor or cardiomyocyte-like phenotypes is of substantial therapeutic interest. Metadichol, a nanoemulsion of long-chain C26–C30 plant-derived aliphatic alcohols, has previously demonstrated pleiotropic biological activities including modulation of nuclear receptors, induction of Yamanaka reprogramming factors, and activation of anti-aging pathways across multiple human cell types. This study presents new quantitative reverse-transcription polymerase chain reaction data characterizing the dose-dependent effects of Metadichol on thirteen cardiac progenitor, pluripotency, and anti-aging genes in human cardiac fibroblasts, and integrates these findings with published data from normal human dermal fibroblasts and human embryonic stem cells.
Methods: Human cardiac fibroblasts were treated with Metadichol at 1 pg/mL, 100 pg/mL, 1 ng/mL, and 100 ng/mL for 24 hours. Gene expression was quantified by SYBR Green quantitative reverse-transcription polymerase chain reaction (Bio-Rad CFX Maestro) using beta-actin as reference, applying the 2^(−ΔΔCt) method. Thirteen target genes were assessed: OCT-4, NANOG, and SOX2 (pluripotency); c-Kit, GATA4, NKX2.5, ISL1, MEF2C, TBX5, MESP1, and WT1 (cardiac progenitor); and TERT and ESRRG (anti-aging and nuclear receptor).
Results: Metadichol treatment induced robust, dose-dependent upregulation with peak induction at 1 ng/mL: c-Kit (12.23-fold), GATA4 (11.51-fold), NKX2.5 (11.48-fold), OCT-4 (10.36-fold), SOX2 (10.21-fold), NANOG (5.86-fold), TERT (4.47-fold), and ESRRG (3.76-fold). TBX5 and MESP1 were selectively downregulated at all doses (0.08- and 0.01-fold minimum, respectively). Correlation analysis revealed strong co-regulation within two gene clusters: pluripotency factors (OCT-4, SOX2, and NANOG; r > 0.95) and cardiac transcription factors (GATA4, NKX2.5, and c-Kit; r > 0.90). The optimal dose in human cardiac fibroblasts (1 ng/mL) differed from that in normal human dermal fibroblasts (100 pg/mL), reflecting cell-type-specific transcriptional thresholds (see Figures 1–4).
Conclusions: Metadichol simultaneously activates pluripotency and cardiac identity gene networks in human cardiac fibroblasts without viral vectors or genetic modification, positioning it as a novel small-molecule cardiac reprogramming agent.