(58) Lycopene Enhances Epigenetic Reprogramming and Zygotic Genome Activation in Porcine Parthenogenetically Activated and Somatic Cell Nuclear Transfer Embryo Development

Ji Hyeon Yun¹; Hyo-Gu Kang¹; Eun Young Choi¹; Se-Been Jeon¹; Min Ju Kim¹; Pil-Soo Jeong¹; Bong-Seok Song¹; Kyungjun Uh¹; Sun-Uk Kim^{1, 2}; Bo-Woong Sim^{1, 2}

1. Futuristic Animal Resource & Research Center (FARRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB), Chungcheongbuk-do 28116, Republic of Korea

2. Department of Functional Genomics, University of Science and Technology (UST), Daejeon 305-333, Republic of Korea

Abstract Text:

Pigs are a valuable model for disease research due to their anatomical and physiological similarities to humans, with somatic cell nuclear transfer (SCNT) commonly used to create such models. However, a major limitation of SCNT is that incomplete somatic cell reprogramming causes epigenetic abnormalities, such as DNA and histone methylation defects, resulting in disrupted zygotic genome activation (ZGA) and reduced embryo viability. Lycopene, a red carotenoid, has strong antioxidant properties and is an effective scavenger of free radicals. Thus, this study investigated whether the addition of lycopene to the culture medium of parthenogenetically activated (PA) and SCNT embryos during in vitro culture (IVC) could improve embryonic development and epigenetic stability. We cultured porcine PA embryos in IVC medium supplemented with varying concentrations of lycopene (0, 0.02, 0.2, and 2 µM) for 6 days to determine the optimal concentration. Supplementation with 0.2 µM lycopene significantly enhanced developmental competence, demonstrated by increased 4–5 cell cleavage rates, blastocyst rates, and total and trophectoderm (TE) cell numbers, along with reduced apoptosis rates and apoptotic cell numbers compared to the control group. Similarly, in SCNT embryos, lycopene treatment improved the 4–5 cell cleavage rates, blastocyst rates, and cell survival rates and increased the number of total and TE cells. This lycopene treatment in PA and SCNT embryos also reduced intracellular ROS levels and upregulated the expression of antioxidant enzyme-related genes (CAT, SOD1, SOD2, and HO-1) in 4-cell embryos and blastocysts. Lycopene treatment enhanced mitochondrial membrane potential and autophagy in PA and SCNT 4-cell embryos. Lycopene treatment in PA and SCNT embryos significantly reduced the levels of H3K4me3, H3K9me3, and 5mC in 4-cell embryos and blastocysts. This lycopene treatment also downregulated the expression levels of methyltransferase-related genes (ASH2L, SUV39H2, DNMT1, DNMT3A, and DNMT3B) while upregulating the expression levels of ZGA-related genes (ZSCAN4, UBTFL1, SUPT4H1, MYC, and ELOA). These results suggest that lycopene treatment during IVC enhances mitochondrial function and autophagic activity, and regulates epigenetic reprogramming, thereby promoting ZGA and improving the embryonic developmental potential of porcine PA and SCNT embryos. This research was supported by the Korea Research Institute of Bioscience and Biotechnology (KRIBB) Research Initiative Program (KGM4252533, KGM5382531), Republic of Korea.