A circular RNA map for human induced pluripotent stem cells of foetal origin
Mario Barilani,
Alessandro Cherubini,
Valeria Peli,
Francesca Polveraccio,
Valentina Bollati,
Federica Guffanti,
Alessandro Del Gobbo,
Cristiana Lavazza,
Silvia Giovanelli,
Nicola Elvassore,
Lorenza Lazzari
Affiliations
Mario Barilani
Laboratory of Regenerative Medicine – Cell Factory, Department of Transfusion Medicine and Haematology, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Via F. Sforza 35, 20122 Milano, Italy; EPIGET Lab, Department of Clinical Sciences and Community Health, Università degli Studi di Milano, Milan, Italy; Department of Industrial Engineering, University of Padova, Padova, Italy
Alessandro Cherubini
Laboratory of Regenerative Medicine – Cell Factory, Department of Transfusion Medicine and Haematology, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Via F. Sforza 35, 20122 Milano, Italy
Valeria Peli
Laboratory of Regenerative Medicine – Cell Factory, Department of Transfusion Medicine and Haematology, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Via F. Sforza 35, 20122 Milano, Italy
Francesca Polveraccio
Laboratory of Regenerative Medicine – Cell Factory, Department of Transfusion Medicine and Haematology, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Via F. Sforza 35, 20122 Milano, Italy; Division of Cancer and Genetics, School of Medicine, Cardiff University, Cardiff, United Kingdom
Valentina Bollati
EPIGET Lab, Department of Clinical Sciences and Community Health, Università degli Studi di Milano, Milan, Italy
Federica Guffanti
Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Milan, Italy
Alessandro Del Gobbo
Division of Pathology, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Italy
Cristiana Lavazza
Laboratory of Regenerative Medicine – Cell Factory, Department of Transfusion Medicine and Haematology, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Via F. Sforza 35, 20122 Milano, Italy
Silvia Giovanelli
Milano Cord Blood Bank, Department of Transfusion Medicine and Haematology, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Milano, Italy
Nicola Elvassore
Department of Industrial Engineering, University of Padova, Padova, Italy; Shanghai Institute for Advanced Immunochemical Studies (SIAIS), ShanghaiTech University, Shanghai, China; Venetian Institute of Molecular Medicine, Padova, Italy; Stem Cells & Regenerative Medicine Section, UCL Great Ormond Street Institute of Child Health, London, United Kingdom
Lorenza Lazzari
Laboratory of Regenerative Medicine – Cell Factory, Department of Transfusion Medicine and Haematology, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Via F. Sforza 35, 20122 Milano, Italy; Corresponding author.
Background: Adult skin fibroblasts represent the most common starting cell type used to generate human induced pluripotent stem cells (F-hiPSC) for clinical studies. Yet, a foetal source would offer unique advantages, primarily the absence of accumulated somatic mutations. Herein, we generated hiPSC from cord blood multipotent mesenchymal stromal cells (MSC-hiPSC) and compared them with F-hiPSC. Assessment of the full activation of the pluripotency gene regulatory network (PGRN) focused on circular RNA (circRNA), recently proposed to participate in the control of pluripotency. Methods: Reprogramming was achieved by a footprint-free strategy. Self-renewal and pluripotency of cord blood MSC-hiPSC were investigated in vitro and in vivo, compared to parental MSC, to embryonic stem cells and to F-hiPSC. High-throughput array-based approaches and bioinformatics analyses were applied to address the PGRN. Findings: Cord blood MSC-hiPSC successfully acquired a complete pluripotent identity. Functional comparison with F-hiPSC showed no differences in terms of i) generation of mesenchymal-like derivatives, ii) their subsequent adipogenic, osteogenic and chondrogenic commitment, and iii) their hematopoietic support ability. At the transcriptional level, specific subsets of mRNA, miRNA and circRNA (n = 4,429) were evidenced, casting a further layer of complexity on the PGRN regulatory crosstalk. Interpretation: A circRNA map of transcripts associated to naïve and primed pluripotency is provided for hiPSC of clinical-grade foetal origin, offering insights on still unreported regulatory circuits of the PGRN to consider for the optimization and development of efficient differentiation protocols for clinical translation. Funding: This research was funded by Ricerca Corrente 2012–2018 by the Italian Ministry of Health.
