Chemical modulation of transcriptionally enriched signaling pathways to optimize the conversion of fibroblasts into neurons
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Direct conversion of human somatic fibroblasts into induced neurons (iNs) allows for the generation of functional neurons while bypassing any stem cell intermediary stages. Although iN technology has an enormous potential for modeling age-related diseases, as well as therapeutic approaches, the technology faces limitations due to variable conversion efficiencies and a lack of thorough understanding of the signaling pathways directing iN conversion. Here, we introduce a new all-in-one inducible lentiviral system that simplifies fibroblast transgenesis for the two pioneer transcription factors, Ngn2 and Ascl1, and markedly improves iN yields. Further, our timeline RNA-Seq data across the course of conversion has identified signaling pathways that become transcriptionally enriched during iN conversion. Small molecular modulators were identified for four signaling pathways that reliably increase the yield of iNs. Taken together, these advances provide an improved toolkit for iN technology and new insight into the mechanisms influencing direct iN conversion.
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Fibroblast-derived iNs preserve substantial signatures of human aging, including transcriptomic, nuclear pore, mitochondrial, epigenetic, and other aspects of aging.
“Fibroblast-derived iNs preserve substantial signatures of human aging, including transcriptomic, nuclear pore, mitochondrial, epigenetic, and other aspects of aging (Herdy et al., 2019; Huh et al., 2016; Kim et al., 2018; Mertens et al., 2015a; Traxler et al., 2019).”
From Age-dependent instability of mature neuronal fate in induced neurons from Alzheimer’s patients (2021) · cites this paper as Herdy et al., 2019
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- Authors
- Herdy, J., Schafer, S., Kim, Y., Ansari, Z., Zangwill, D., Ku, M., Paquola, A., Lee, H., Mertens, J., & Gage, F. H.
- Affiliations
- Salk Institute for Biological Studies (US)
- University Hospital Freiberg, University of Freiberg, Breisgau, Germany
- Lieber Institute for Brain Development, Baltimore, United States
- Laboratory of Genetics, The Salk Institute for Biological Studies, La Jolla, United States
- Corresponding author
- Gage, Fred H
- Venue
- eLife
- Publisher
- eLife Sciences Publications, Ltd
- Published
- 2019-05-17
- Volume
- 8
- ISSN
- 2050-084X (ISSN-L)
- PMC
- PMC6524968
- OpenAlex
- W2946301480
- Cited by
- 64
- References
- 60
- Citations by year
- 2026: 8 · 2025: 5 · 2024: 7 · 2023: 8 · 2022: 6 · 2021: 12 · 2020: 13 · 2019: 5
- FWCI
- 2.98
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- eLife (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Austrian Science Fund
- Paul G. Allen Family Foundation
- National Institute on Aging (R01-AG056306-01, K99-AG056679-01)
- JPB Foundation
- Glenn Family Foundation
- American Federation for Aging Research
- Leona M. and Harry B. Helmsley Charitable Trust (2012- PG-MED002)
- Annette Merle-Smith CIRM (TR2-01778)
- G Harold and Leila Y. Mathers Foundation
- Brain and Behavior Research Foundation (2014 NARSAD Young Investigator Grant)
- National Cancer Institute (P30 CA014195)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
- Keywords
- induced neurons, Ngn2, ASCL1, lentiviral transduction, small molecule modulators, signaling pathways, conversion efficiency, neuronal differentiation
- MeSH terms
- Adolescent, Adult, Aged, Aged, 80 and over, Cells, Cultured, Child, Child, Preschool, Fibroblasts, Humans, Infant, Infant, Newborn, Middle Aged, Nerve Tissue Proteins, Neurons, Signal Transduction, Sequence Analysis, RNA, Gene Transfer Techniques, Gene Expression Profiling, Basic Helix-Loop-Helix Transcription Factors, Basic Helix-Loop-Helix Proteins, Cell Transdifferentiation, Young Adult
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