DNA barcoding technology for tracking cell lineage of multinucleated osteoclasts

2023 Fiscal Year Final Research Report

• Project/Area Number: 22K19289
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 43:Biology at molecular to cellular levels, and related fields
• Research Institution: Osaka University
• Principal Investigator: Yahara Yasuhito 大阪大学, 大学院生命機能研究科, 准教授 (60456390)
• Project Period (FY): 2022-06-30 – 2024-03-31
• Keywords: 破骨細胞 / 細胞融合 / 系譜解析 / 胎児卵黄嚢 / 造血幹細胞

Outline of Final Research Achievements

Osteoclasts are multinucleated cells that form through the fusion of precursor cells. This study aimed to develop foundational technology to track the fusion process of osteoclast precursors with distinct genetic profiles. Mice were engineered to express three different fluorescent proteins, labeling osteoclasts from varied origins. Using in situ hybridization (ISH), we detected the RNA molecules of these fluorescent proteins, thereby identifying the origins of the osteoclasts. Moreover, we established mice that specifically express molecular barcodes, guide RNA (gRNA), and Cas9 protein in osteoclas. This allowed for the analysis of cell lineage based on the genetic barcodes embedded in these precursors. This research introduces a novel technology for analyzing the diversity of precursor cells contributing to a single mature osteoclast and the regularity of their fusion.

Free Research Field

骨代謝学

Academic Significance and Societal Importance of the Research Achievements

本研究は、一つの成熟破骨細胞を構成する前駆細胞の多様性や細胞融合の規則性を網羅的に解析する新技術の創出に貢献した。新たに樹立した多系統破骨細胞の系譜解析マウスは、従来の解析法では観察が困難であった細胞同士の融合現象やその生理的意義に関する洞察を深め、多核細胞形成ダイナミクスと、その多様性構築メカニズムの解明を可能にする画期的な手法を提供した。この新手法は、破骨細胞の病的活性化や機能不全における細胞融合メカニズムの解明に向けた研究成果に繋がると期待される。