Vasculogenic skin reprogramming requires TET-mediated gene demethylation in fibroblasts for rescuing impaired perfusion in diabetes
Tissue nanotransfection (TNT) topically delivers Etv2, Foxc2, and Fli1 (EFF) plasmids increasing vasculogenic fibroblasts (VF) and promoting vascularization in ischemic murine skin. Human dermal fibroblasts respond to EFF nanoelectroporation with elevated expression of endothelial genes in vitro, which is linked to increased ten-eleven translocase 1/2/3 (TET) expression. Single cell RNA sequencing dependent validation of VF induction reveals a TET-dependent transcript signature. TNT<sub>EFF</sub> also induces TET expression in vivo, and fibroblast-specific EFF overexpression leads to VF-transition, with TET-activation correlating with higher 5-hydroxymethylcytosine (5-hmC) levels in VF. VF emergence requires TET-dependent demethylation of endothelial genes in vivo, enhancing VF abundance and restoring perfusion in diabetic ischemic limbs. TNT<sub>EFF</sub> improves perfusion and wound closure in diabetic mice, while increasing VF in cultured human skin explants. Suppressed in diabetes, TET1/2/3 play a critical role in TNT-mediated VF formation which supports de novo blood vessel development to rescue diabetic ischemic tissue.
Authors
- Ravichand Palakurti
- Yashika Rustagi (ORCID: https://orcid.org/0000-0003-1122-941X)
- Priyanka Verma (ORCID: https://orcid.org/0000-0002-3035-4964)
- Sashwati Roy (ORCID: https://orcid.org/0000-0002-9995-4917)
- Manishekhar Kumar (ORCID: https://orcid.org/0000-0002-5364-2520)
- Savita Khanna (ORCID: https://orcid.org/0000-0001-6661-9293)
- Sujit K. Mohanty (ORCID: https://orcid.org/0000-0001-9559-4638)
- Sumit Singh Verma (ORCID: https://orcid.org/0000-0002-3559-158X)
- Mervin C. Yöder (ORCID: https://orcid.org/0000-0003-4354-7631)
- Chandan K. Sen (ORCID: https://orcid.org/0000-0003-3151-5202)
- Surya Gnyawali (ORCID: https://orcid.org/0000-0001-8980-4889)
- Sedat Kaçar (ORCID: https://orcid.org/0000-0002-0671-8529)
- Pradipta K. Banerjee (ORCID: https://orcid.org/0009-0000-7835-9345)
- Subhadip Ghatak (ORCID: https://orcid.org/0000-0001-5641-3902)
- Ajay Kumar Sahi (ORCID: https://orcid.org/0000-0002-1035-8562)
- Edward Hernandez (ORCID: https://orcid.org/0000-0001-6638-6303)
- Mohamed S. El Masry (ORCID: https://orcid.org/0000-0002-2686-1509)
- Rajneesh Srivastava (ORCID: https://orcid.org/0000-0003-1093-2082)
- Kanhaiya Singh (ORCID: https://orcid.org/0000-0002-4126-9618)
- J. Peter Rubin
Institutions
- University of Pittsburgh (US)
- McGowan Institute for Regenerative Medicine (US)
- Indiana University School of Medicine
- Indiana University – Purdue University Indianapolis (US)
Publication Details
- Journal
- Nature Communications
- Published
- 2024-11-27
- DOI
- https://doi.org/10.1038/s41467-024-54385-w
- Citations
- 12
- Primary Topic
- Single-cell and spatial transcriptomics
- Type
- article
- Field-Weighted Citation Impact
- 2.81
Funders
- U.S. Department of Defense
- John Templeton Foundation
- Eli Lilly and Company
- Yale University
- Lilly Endowment
- National Institute of Diabetes and Digestive and Kidney Diseases
- Congressionally Directed Medical Research Programs