Advertisement

ResearchIn-Press PreviewAngiogenesisVascular biology Open Access | 10.1172/JCI188559

A smooth muscle cell lncRNA controls angiogenesis in chronic limb-threatening ischemia through miR-143-3p/HHIP signaling

Ming Zhai,1 Anurag Jamaiyar,1 Jun Qian,1 Winona W. Wu,1 Emre Bektik,1 Vinay Randhawa,1 Camila De Oliveira Vaz,1 Arvind K. Pandey,1 Akm Khyrul Wara,1 Madhur Sachan,1 Yi Hu,1 Jéssica L. Garcia,1 Claire E. Alford,1 Terence E. Ryan,2 Wenhui Peng,3 and Mark W. Feinberg1

1Cardiovascular Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, United States of America

2Department of Applied Physiology and Kinesiology, University of Florida, Gainesville, United States of America

3Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China

Find articles by Zhai, M. in: PubMed | Google Scholar

1Cardiovascular Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, United States of America

2Department of Applied Physiology and Kinesiology, University of Florida, Gainesville, United States of America

3Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China

Find articles by Jamaiyar, A. in: PubMed | Google Scholar |

1Cardiovascular Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, United States of America

2Department of Applied Physiology and Kinesiology, University of Florida, Gainesville, United States of America

3Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China

Find articles by Qian, J. in: PubMed | Google Scholar

1Cardiovascular Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, United States of America

2Department of Applied Physiology and Kinesiology, University of Florida, Gainesville, United States of America

3Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China

Find articles by Wu, W. in: PubMed | Google Scholar

1Cardiovascular Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, United States of America

2Department of Applied Physiology and Kinesiology, University of Florida, Gainesville, United States of America

3Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China

Find articles by Bektik, E. in: PubMed | Google Scholar

1Cardiovascular Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, United States of America

2Department of Applied Physiology and Kinesiology, University of Florida, Gainesville, United States of America

3Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China

Find articles by Randhawa, V. in: PubMed | Google Scholar

1Cardiovascular Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, United States of America

2Department of Applied Physiology and Kinesiology, University of Florida, Gainesville, United States of America

3Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China

Find articles by De Oliveira Vaz, C. in: PubMed | Google Scholar

1Cardiovascular Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, United States of America

2Department of Applied Physiology and Kinesiology, University of Florida, Gainesville, United States of America

3Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China

Find articles by Pandey, A. in: PubMed | Google Scholar

1Cardiovascular Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, United States of America

2Department of Applied Physiology and Kinesiology, University of Florida, Gainesville, United States of America

3Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China

Find articles by Wara, A. in: PubMed | Google Scholar

1Cardiovascular Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, United States of America

2Department of Applied Physiology and Kinesiology, University of Florida, Gainesville, United States of America

3Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China

Find articles by Sachan, M. in: PubMed | Google Scholar

1Cardiovascular Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, United States of America

2Department of Applied Physiology and Kinesiology, University of Florida, Gainesville, United States of America

3Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China

Find articles by Hu, Y. in: PubMed | Google Scholar

1Cardiovascular Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, United States of America

2Department of Applied Physiology and Kinesiology, University of Florida, Gainesville, United States of America

3Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China

Find articles by Garcia, J. in: PubMed | Google Scholar

1Cardiovascular Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, United States of America

2Department of Applied Physiology and Kinesiology, University of Florida, Gainesville, United States of America

3Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China

Find articles by Alford, C. in: PubMed | Google Scholar

1Cardiovascular Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, United States of America

2Department of Applied Physiology and Kinesiology, University of Florida, Gainesville, United States of America

3Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China

Find articles by Ryan, T. in: PubMed | Google Scholar |

1Cardiovascular Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, United States of America

2Department of Applied Physiology and Kinesiology, University of Florida, Gainesville, United States of America

3Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China

Find articles by Peng, W. in: PubMed | Google Scholar |

1Cardiovascular Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, United States of America

2Department of Applied Physiology and Kinesiology, University of Florida, Gainesville, United States of America

3Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China

Find articles by Feinberg, M. in: PubMed | Google Scholar |

Published August 28, 2025 - More info

J Clin Invest. https://doi.org/10.1172/JCI188559.
Copyright © 2025, Zhai et al. This work is licensed under the Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
Published August 28, 2025 - Version history
View PDF
Abstract

Peripheral artery disease (PAD) often advances to chronic limb-threatening ischemia (CLTI), resulting in severe complications such as limb amputation. Despite the potential of therapeutic angiogenesis, the mechanisms of cell-cell communication and transcriptional changes driving PAD are not fully understood. Profiling long non-coding RNAs (lncRNAs) from gastrocnemius muscles of human subjects with or without CLTI revealed that a vascular smooth muscle cell (SMC)-enriched lncRNA CARMN, was reduced with CLTI. This study explored how a SMC lncRNA-miRNA signaling axis regulates angiogenesis in limb ischemia. CARMN knockout (KO) mice exhibited reduced capillary density and impaired blood flow recovery and tissue necrosis following limb ischemia. We found that CARMN KO SMC supernatants inhibited endothelial cell (EC) proliferation, spheroid sprouting, and network formation. RNA-sequencing identified downregulation of the Hedgehog signaling pathway in CARMN KO models and revealed that CARMN regulates this pathway through its downstream miRNA, miR-143-3p, which targets Hedgehog-interacting protein (HHIP), an antagonist of Hedgehog signaling. Delivery of HHIP-specific siRNA or miR-143-3p mimics rescued EC angiogenic defects and improved blood flow recovery in both CARMN KO and WT mice. These findings underscore the critical role of CARMN in modulating angiogenesis through the miR-143-3p-HHIP-Hedgehog signaling axis, providing insights into SMC-EC interactions and potential therapeutic strategies for CLTI.

Graphical Abstract
graphical abstract
Supplemental material

View Unedited blot and gel images

View Supplemental Table 1

View Supplemental Table 2

View

Version history
  • Version 1 (August 28, 2025): In-Press Preview
Advertisement
Advertisement