Academic Journal
Developmental stage-dependent cellular plasticity governs β-cell reprogramming efficiency in human fibroblasts.
| Title: | Developmental stage-dependent cellular plasticity governs β-cell reprogramming efficiency in human fibroblasts. |
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| Authors: | Raj N; Laboratory for Stem Cell Engineering and Regenerative Medicine, Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati, Assam, 781039, India. Electronic address: naveenraj@iitg.ac.in., Panneerselvam DS; Laboratory for Stem Cell Engineering and Regenerative Medicine, Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati, Assam, 781039, India. Electronic address: p.dhaya@iitg.ac.in., Thummer RP; Laboratory for Stem Cell Engineering and Regenerative Medicine, Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati, Assam, 781039, India. Electronic address: rthu@iitg.ac.in. |
| Source: | Biochemical and biophysical research communications [Biochem Biophys Res Commun] 2026 Sep 03; Vol. 829, pp. 154188. Date of Electronic Publication: 2026 Jun 23. |
| Publication Type: | Journal Article |
| Language: | English |
| Journal Info: | Publisher: Elsevier Country of Publication: United States NLM ID: 0372516 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1090-2104 (Electronic) Linking ISSN: 0006291X NLM ISO Abbreviation: Biochem Biophys Res Commun Subsets: MEDLINE |
| Imprint Name(s): | Publication: <2002- >: San Diego, CA : Elsevier Original Publication: New York, Academic Press. |
| MeSH Terms: | Fibroblasts*/cytology , Fibroblasts*/metabolism , Insulin-Secreting Cells*/cytology , Insulin-Secreting Cells*/metabolism , Cellular Reprogramming* , Cell Plasticity*, Lentivirus/genetics ; Trans-Activators/genetics ; Trans-Activators/metabolism ; Nerve Tissue Proteins/genetics ; Nerve Tissue Proteins/metabolism ; Homeodomain Proteins/genetics ; Homeodomain Proteins/metabolism ; Basic Helix-Loop-Helix Proteins/genetics ; Basic Helix-Loop-Helix Proteins/metabolism ; Humans ; Cells, Cultured ; Cell Differentiation |
| Abstract: | Direct reprogramming of somatic cells into alternative lineages, bypassing a pluripotent intermediate state, represents a promising strategy to reduce the risks of tumorigenesis and genomic instability associated with induced pluripotent stem cell-based approaches. Although β-cell-like cells have been efficiently generated from developmentally related endodermal sources such as acinar cells, ductal cells and α-cells, their derivation from phylogenetically distant cell types, such as fibroblasts, remains challenging. In this study, we employed lentiviral-mediated reprogramming using a polycistronic construct encoding the pancreatic transcription factors Neurog3, Pdx1 and MafA to enable β-cell-like formation from human fibroblasts. To assess the influence of developmental stage on reprogramming efficiency, fibroblasts derived from neonatal, juvenile and adult stages were analyzed comparatively. Transgene delivery, expression and functional effects were validated by reporter gene analysis, microscopy, flow cytometry, RT-qPCR and immunoblotting. Reprogrammed cells derived from neonatal fibroblasts exhibited the highest reprogramming efficiency, as demonstrated by robust induction of INS and MAFA expression, along with concurrent downregulation of fibroblast marker PRRX1. In contrast, reprogrammed cells derived from adult fibroblasts displayed markedly reduced responsiveness. Flow cytometry analysis revealed that approximately 38% of reprogrammed cells derived from neonatal fibroblasts acquired an INS-positive phenotype. Notably, reprogrammed cells derived from juvenile fibroblasts exhibited a higher proportion of GCG-expressing cells compared to those derived from neonatal fibroblasts, indicating a more pronounced heterogeneous endocrine-like state characteristic of intermediate stages of pancreatic differentiation in the former compared to the latter. Collectively, these findings demonstrate that the developmental stage significantly influences reprogramming competence, with neonatal fibroblasts representing a more permissive cell source for β-cell-like conversion. These results highlight developmental stage-dependent cellular plasticity as a key determinant of reprogramming efficiency and reveal the optimal cell sources for β-cell generation. (Copyright © 2026. Published by Elsevier Inc.) |
| Competing Interests: | Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. |
| Contributed Indexing: | Keywords: Cellular plasticity; Direct reprogramming; Human fibroblasts; Lentiviral transduction; Transcription factors; β-cell–like |
| Substance Nomenclature: | 0 (Trans-Activators) 0 (Nerve Tissue Proteins) 0 (Homeodomain Proteins) 0 (Basic Helix-Loop-Helix Proteins) 0 (NEUROG3 protein, human) 0 (pancreatic and duodenal homeobox 1 protein) |
| Entry Date(s): | Date Created: 20260623 Date Completed: 20260716 Latest Revision: 20260716 |
| Update Code: | 20260717 |
| DOI: | 10.1016/j.bbrc.2026.154188 |
| PMID: | 42335655 |
| Database: | MEDLINE |
| ISSN: | 1090-2104 |
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| DOI: | 10.1016/j.bbrc.2026.154188 |