Preventing alloimmune rejection using circular RNA FSCN1-silenced dendritic cells in heart transplantation

Bowen Wang; Qinfeng Zhou; Toni Li; Shuailong Li; Adam Greasley; Anton Skaro; Douglas Quan; Weiping Min; Kexiang Liu; Xiufen Zheng

doi:10.1016/j.healun.2021.03.025

Preventing alloimmune rejection using circular RNA FSCN1-silenced dendritic cells in heart transplantation

J Heart Lung Transplant. 2021 Jul;40(7):584-594. doi: 10.1016/j.healun.2021.03.025. Epub 2021 Apr 8.

Authors

Bowen Wang¹, Qinfeng Zhou², Toni Li³, Shuailong Li¹, Adam Greasley⁴, Anton Skaro², Douglas Quan², Weiping Min⁵, Kexiang Liu⁶, Xiufen Zheng⁷

Affiliations

¹ Department of Cardiovascular Surgery, The Second Affiliated Hospital, Jilin University, Changchun, China; Department of Pathology and Laboratory Medicine, Western University, London, Ontario Canada.
² Department of Surgery, Western University, London, Ontario Canada.
³ Department of Pathology and Laboratory Medicine, Western University, London, Ontario Canada; School of Medicine, Queen's University, Kingston, Canada.
⁴ Department of Pathology and Laboratory Medicine, Western University, London, Ontario Canada.
⁵ Department of Pathology and Laboratory Medicine, Western University, London, Ontario Canada; Department of Surgery, Western University, London, Ontario Canada; Lawson Health Research Institute, London, Ontario Canada; Department of Oncology, Western University, London, Ontario Canada.
⁶ Department of Cardiovascular Surgery, The Second Affiliated Hospital, Jilin University, Changchun, China.
⁷ Department of Pathology and Laboratory Medicine, Western University, London, Ontario Canada; Department of Surgery, Western University, London, Ontario Canada; Lawson Health Research Institute, London, Ontario Canada; Department of Oncology, Western University, London, Ontario Canada. Electronic address: xzheng26@uwo.ca.

PMID: 34052126
DOI: 10.1016/j.healun.2021.03.025

Abstract

Background: While heart transplantation is used as a standard treatment for heart failure, transplant rejection continues to pose a challenge. Recent evidence has shown that circular RNA (circRNA) is a new type of gene regulator in cell development. Our aim was to demonstrate that treatment with tolerogenic dendritic cells (Tol-DCs) generated by circular RNA FSCN1 (circFSCN1) silencing could prevent alloimmune rejection and prolong heart graft survival in heart transplantation.

Methods: Bone marrow-derived DCs were transfected with circFSCN1 siRNA in vitro. The circFSCN1 level was measured by qRT-PCR. DC maturation was determined by flow cytometry. Mixed lymphocyte reactions (MLRs) were conducted to assess the function of DCs to activate T cells and to generate regulatory T cells (Tregs). In situ RNA hybridization and fluorescent microscopy were performed to detect the distribution of circFSCN1 in DCs. A heterotopic allogeneic murine heart transplantation was conducted where recipients were pre-treated with donor derived circFSCN1-silenced Tol-DCs. Heartbeat was monitored to assess immune rejection.

Results: Exonic circFSCN1 was highly expressed in the cytoplasm of mature DCs. Knockdown of circFSCN1 using siRNA arrested DCs at an immature state, impaired DC's ability to activate T cells and enhanced Treg generation. Treatment with circFSCN1-silenced Tol-DCs prevented alloimmune rejection, prolonged allograft survival, reduced fibrosis, and induced Tregs in vivo.

Conclusions: Knockdown of circFSCN1 induces Tol-DCs and treatment with these Tol-DCs prevents alloimmune rejection and prolongs allograft survival. This is a promising therapeutic target to combat transplant rejection in heart transplantation and increases our understanding of circRNA in the immune system.

Keywords: circFSCN1; circular RNA; heart transplantation; immune rejection; tolerogenic DCs.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Dendritic Cells / immunology*
Disease Models, Animal
Gene Expression Regulation*
Graft Rejection / genetics
Graft Rejection / immunology
Graft Rejection / prevention & control*
Graft Survival
Heart Transplantation / adverse effects*
Immune Tolerance / genetics*
Lymphocyte Activation
Male
Mice
Mice, Inbred BALB C
Mice, Inbred C57BL
Microfilament Proteins / biosynthesis
Microfilament Proteins / genetics*
RNA, Circular / genetics*
Receptors, Odorant / biosynthesis
Receptors, Odorant / genetics*
T-Lymphocytes, Regulatory / immunology
Transplantation, Homologous

Substances

Microfilament Proteins
RNA, Circular
Receptors, Odorant
fascin1 protein, mouse

Abstract

Publication types

MeSH terms

Substances

Grants and funding