Conversion of human fibroblasts to angioblast-like progenitor cells

Abstract

Lineage conversion of one somatic cell type to another is an attractive approach for generating specific human cell types. Lineage conversion can be direct, in the absence of proliferation and multipotent progenitor generation, or indirect, by the generation of expandable multipotent progenitor states. We report the development of a reprogramming methodology in which cells transition through a plastic intermediate state, induced by brief exposure to reprogramming factors, followed by differentiation. We use this approach to convert human fibroblasts to mesodermal progenitor cells, including by non-integrative approaches. These progenitor cells demonstrated bipotent differentiation potential and could generate endothelial and smooth muscle lineages. Differentiated endothelial cells exhibited neo-angiogenesis and anastomosis in vivo. This methodology for indirect lineage conversion to angioblast-like cells adds to the armamentarium of reprogramming approaches aimed at the study and treatment of ischemic pathologies.

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Figure 1: Differentiation of hPSCs into mesodermal progenitor and endothelial cells.
Figure 2: Conversion of human fibroblasts into mesodermal progenitors and endothelial cells by retroviral approaches.
Figure 3: Conversion of human fibroblasts to mesodermal progenitors and endothelial cells by non-integrative approaches.
Figure 4: Converted endothelial cells are functional in vivo.

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  • 07 February 2020

    "An amendment to this paper has been published and can be accessed via a link at the top of the paper."

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Acknowledgements

We are thankful to Y. Zheng for his expertise and assistance with sorting procedures, C. Maiza for his expertise and assistance with in vivo procedures and M. Schwarz for administrative support. L.K. was partially supported by the California Institute for Regenerative Medicine. E.N. was partially supported by an F.M. Kirby Foundation postdoctoral fellowship. A.M.D. was supported by the Helmsley Foundation. L.C.L., R.D.T., F.S.B. and J.F.L. are supported by the California Institute for Regenerative Medicine (CL1-00502, RT1-01108, TR1-01250, RN2-00931), US National Institutes of Health (R33MH087925), US National Institutes of Health/National Institute Child Health and Human Development K12 Career Development Award (L.C.L.), Hartwell Foundation (L.C.L., R.D.T., F.S.B.), Millipore Foundation (J.F.L.) and Esther O'Keefe Foundation (J.F.L.). Work in the laboratory of F.H.G. was supported by the JPB Foundation, G. Harold and Leila Y. Mathers Charitable Foundation and Ellison Medical Foundation. Work in the laboratory of J.C.I.B. was supported by grants from Fundacion Cellex, the G. Harold and Leila Y. Mathers Charitable Foundation, the Leona M. and Harry B. Helmsley Charitable Trust, Sanofi, Ministerio de Economia y Competitividad (PLE2009-0100), Instituto de Salud Carlos III (ISCIII), Terapia Celular (TerCel) (RD06/0010/0016) and Fondo Europeo de Desarrollo Regional (FEDER).

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L.K., I.S.-M., E.N. and J.C.I.B. designed all experiments. I.S.-M., E.N. and J.C.I.B. wrote the manuscript. L.K., I.S.-M., K.M., E.N. and A.A. performed and analyzed all experiments. C.P., C.V.-C., F.B., E.N., I.D. and J.-M.H. performed in vivo experiments. K.M. was responsible for all cell culture–related work. M.L., A.M.D. and F.H.G. provided microRNA constructs and reagents. J.P., Y.X., S.R., I.D., N.M., C.R., A.M.D. and F.H.G. contributed to the overall design of the project. F.S.B., R.D.T., J.F.L. and L.C.L. performed and analyzed genome-wide array DNA methylation and gene expression studies.

Corresponding author

Correspondence to Juan Carlos Izpisua Belmonte.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–12 and Supplementary Tables 1 and 3 (PDF 23185 kb)

Supplementary Table 2

Specific mRNA fold change of the qPCR data summarized in Figures 1–3. (XLSX 54 kb)

Supplementary Data

Detailed mRNA and methylation array data including differentially regulated gene lists. (XLSX 22235 kb)

Generated smooth muscle cells demonstrating contractile capabilities.

(AVI 9125 kb)

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Kurian, L., Sancho-Martinez, I., Nivet, E. et al. Conversion of human fibroblasts to angioblast-like progenitor cells. Nat Methods 10, 77–83 (2013). https://doi.org/10.1038/nmeth.2255

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