One Molecule, Two Fates: Metadichol Co-Activates Pluripotency and Cardiac Gene Networks in Human Cardiac Fibroblasts

Background Cardiovascular disease remains the leading cause of global mortality, with myocardial fibrosis representing a critical barrier to cardiac regeneration. Human cardiac fibroblasts (HCFs) 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 (NRs), induction of Yamanaka reprogramming factors, and activation of anti-aging pathways across multiple human cell types. This study presents new quantitative RT-PCR data characterizing Metadichol's dose-dependent effects on 13 cardiac progenitor, pluripotency, and anti-aging genes in HCFs, and integrates these findings with published data from normal human dermal fibroblasts (NHDFs) and human embryonic stem cells (hESCs). Methods HCFs 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 qRT-PCR (Bio-Rad CFX Maestro) using beta-actin as reference, applying the 2^(-??Ct) method. Thirteen target genes were assessed: OCT-4, NANOG, SOX2 (Yamanaka/pluripotency); c-Kit, GATA4, NKX2.5, ISL1, MEF2C, TBX5, MESP1, WT1 (cardiac progenitor); TERT, ESRRG (anti-aging/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/NANOG, r>0.95) and cardiac TFs (GATA4/NKX2.5/c-Kit, r>0.90). The optimal dose in HCFs (1 ng/mL) differed from NHDFs (100 pg/mL), reflecting cell-type-specific transcriptional thresholds (see Figures 1-4). Conclusions Metadichol simultaneously activates pluripotency and cardiac identity gene networks in HCFs without viral vectors or genetic modification, positioning it as a novel small-molecule cardiac reprogramming agent.