Junior Associate Professor Center for iPS Cell Research and Application (CiRA), Kyoto University Kyoto, Kyoto, Japan
Abstract: Reprogramming peripheral blood mononuclear cells (PBMCs) into induced pluripotent stem cells (iPSCs) is essential for advancing regenerative medicine and disease modeling. However, challenges remain in enhancing efficiency and understanding the molecular mechanisms involved. Synthetic mRNA offers a non-integrative and effective method for reprogramming, yet its full potential has yet to be realized. To improve reprogramming efficiency, we focused on PBMCs and demonstrated that modulation of the p53 pathway significantly enhances reprogramming outcomes by improving cellular survival and reducing stress-induced barriers. This identifies a novel target for optimizing PBMC reprogramming protocols. To further investigate the mechanisms of cellular reprogramming, we utilized human dermal fibroblasts (HDFs) as a model system and implemented an integrative RNA sequencing (RNA-seq) strategy that combines short-read and long-read technologies. Preliminary analyses suggest distinct transcriptional phases during reprogramming, marked by the activation of pluripotency-associated genes, suppression of somatic gene networks, and potential identification of new isoforms and regulatory elements. These insights, while still ongoing, provide a foundational understanding of transcriptional dynamics and support hypotheses for optimizing reprogramming conditions. Together, these findings not only enhance our mechanistic understanding of reprogramming but also establish a robust platform for improving iPSC generation from PBMCs, paving the way for expanded therapeutic applications.
Funding Source: This study was supported by Core Center for iPS Cell Research, AMED (Japan), the iPS Cell Research Fund (Japan), and World Premier International Research Center Initiative (WPI), MEXT (Japan).
