PMID- 37194082 OWN - NLM STAT- MEDLINE DCOM- 20230518 LR - 20230603 IS - 1757-6512 (Electronic) IS - 1757-6512 (Linking) VI - 14 IP - 1 DP - 2023 May 16 TI - CINC-2 and miR-199a-5p in EVs secreted by transplanted Thy1(+) cells activate hepatocytic progenitor cell growth in rat liver regeneration. PG - 134 LID - 10.1186/s13287-023-03346-z [doi] LID - 134 AB - BACKGROUND: Small hepatocyte-like progenitor cells (SHPCs) are hepatocytic progenitor cells that transiently form clusters in rat livers treated with retrorsine (Ret) that underwent 70% partial hepatectomy (PH). We previously reported that transplantation of Thy1(+) cells obtained from D-galactosamine-treated livers promotes SHPC expansion, thereby accelerating liver regeneration. Extracellular vesicles (EVs) secreted by Thy1(+) cells induce sinusoidal endothelial cells (SECs) and Kupffer cells (KCs) to secrete IL17B and IL25, respectively, thereby activating SHPCs through IL17 receptor B (RB) signaling. This study aimed to identify the inducers of IL17RB signaling and growth factors for SHPC proliferation in EVs secreted by Thy1(+) cells (Thy1-EVs). METHODS: Thy1(+) cells isolated from the livers of rats treated with D-galactosamine were cultured. Although some liver stem/progenitor cells (LSPCs) proliferated to form colonies, others remained as mesenchymal cells (MCs). Thy1-MCs or Thy1-LSPCs were transplanted into Ret/PH-treated livers to examine their effects on SHPCs. EVs were isolated from the conditioned medium (CM) of Thy1-MCs and Thy1-LSPCs. Small hepatocytes (SHs) isolated from adult rat livers were used to identify factors regulating cell growth in Thy1-EVs. RESULTS: The size of SHPC clusters transplanted with Thy1-MCs was significantly larger than that of SHPC clusters transplanted with Thy1-LSPCs (p = 0.02). A comprehensive analysis of Thy1-MC-EVs revealed that miR-199a-5p, cytokine-induced neutrophil chemoattractant-2 (CINC-2), and monocyte chemotactic protein 1 (MCP-1) were candidates for promoting SHPC growth. Additionally, miR-199a-5p mimics promoted the growth of SHs (p = 0.02), whereas CINC-2 and MCP-1 did not. SECs treated with CINC-2 induced Il17b expression. KCs treated with Thy1-EVs induced the expression of CINC-2, Il25, and miR-199a-5p. CM derived from SECs treated with CINC-2 accelerated the growth of SHs (p = 0.03). Similarly, CM derived from KCs treated with Thy1-EVs and miR-199a-5p mimics accelerated the growth of SHs (p = 0.007). In addition, although miR-199a-overexpressing EVs could not enhance SHPC proliferation, transplantation of miR-199a-overexpressing Thy1-MCs could promote the expansion of SHPC clusters. CONCLUSION: Thy1-MC transplantation may accelerate liver regeneration owing to SHPC expansion, which is induced by CINC-2/IL17RB signaling and miR-199a-5p via SEC and KC activation. CI - (c) 2023. The Author(s). FAU - Ichinohe, Norihisa AU - Ichinohe N AUID- ORCID: 0000-0003-3723-7418 AD - Department of Tissue Development and Regeneration, Research Institute for Frontier Medicine, Sapporo Medical University School of Medicine, South-1, West-17, Chuo-Ku, Sapporo, 060-8556, Japan. nichi@sapmed.ac.jp. FAU - Tanimizu, Naoki AU - Tanimizu N AD - Department of Tissue Development and Regeneration, Research Institute for Frontier Medicine, Sapporo Medical University School of Medicine, South-1, West-17, Chuo-Ku, Sapporo, 060-8556, Japan. AD - Division of Regenerative Medicine, Center for Stem Cell Biology and Regenerative Medicine, The Institute of Medical Science, University of Tokyo, Tokyo, Japan. FAU - Ishigami, Keisuke AU - Ishigami K AD - Department of Tissue Development and Regeneration, Research Institute for Frontier Medicine, Sapporo Medical University School of Medicine, South-1, West-17, Chuo-Ku, Sapporo, 060-8556, Japan. FAU - Yoshioka, Yusuke AU - Yoshioka Y AD - Division of Molecular and Cellular Medicine, National Cancer Center Research Institute, Tokyo, Japan. AD - Department of Molecular and Cellular Medicine, Institute of Medical Science, Tokyo Medical University, Tokyo, Japan. FAU - Fujitani, Naoki AU - Fujitani N AD - Department of Biochemistry, Sapporo Medical University School of Medicine, Sapporo, Japan. FAU - Ochiya, Takahiro AU - Ochiya T AD - Division of Molecular and Cellular Medicine, National Cancer Center Research Institute, Tokyo, Japan. AD - Department of Molecular and Cellular Medicine, Institute of Medical Science, Tokyo Medical University, Tokyo, Japan. FAU - Takahashi, Motoko AU - Takahashi M AD - Department of Biochemistry, Sapporo Medical University School of Medicine, Sapporo, Japan. FAU - Mitaka, Toshihiro AU - Mitaka T AD - Department of Tissue Development and Regeneration, Research Institute for Frontier Medicine, Sapporo Medical University School of Medicine, South-1, West-17, Chuo-Ku, Sapporo, 060-8556, Japan. tmitaka@sapmed.ac.jp. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20230516 PL - England TA - Stem Cell Res Ther JT - Stem cell research & therapy JID - 101527581 RN - 7535-00-4 (Galactosamine) RN - 0 (MicroRNAs) RN - 0 (MIRN199 microRNA, rat) RN - 0 (Cxcl3 protein, rat) RN - 0 (Chemokines, CXC) SB - IM MH - Animals MH - Rats MH - Cell Proliferation MH - Endothelial Cells MH - *Extracellular Vesicles MH - Galactosamine MH - Hepatocytes/metabolism MH - Liver Regeneration/physiology MH - *MicroRNAs/genetics/metabolism MH - Rats, Inbred F344 MH - Stem Cells/metabolism MH - *Chemokines, CXC/genetics/metabolism PMC - PMC10190025 OTO - NOTNLM OT - CINC-2 OT - Extracellular vesicles OT - Hepatocytic progenitor cells OT - IL17RB signal OT - Small hepatocytes OT - Thy1+ mesenchymal cells OT - miR-199a-5p COIS- The authors indicate no potential conflict of interest. EDAT- 2023/05/17 01:07 MHDA- 2023/05/18 06:42 PMCR- 2023/05/16 CRDT- 2023/05/16 23:38 PHST- 2022/09/26 00:00 [received] PHST- 2023/04/12 00:00 [accepted] PHST- 2023/05/18 06:42 [medline] PHST- 2023/05/17 01:07 [pubmed] PHST- 2023/05/16 23:38 [entrez] PHST- 2023/05/16 00:00 [pmc-release] AID - 10.1186/s13287-023-03346-z [pii] AID - 3346 [pii] AID - 10.1186/s13287-023-03346-z [doi] PST - epublish SO - Stem Cell Res Ther. 2023 May 16;14(1):134. doi: 10.1186/s13287-023-03346-z.