The Role of Circular RNA Derived from Endothelial Cells in Promoting the Formation of Colonies in Procr+ Islet Cell

Yihan Deng

doi:10.54254/2753-8818/2025.AU27351

Theoretical and Natural ScienceOpen access

The Role of Circular RNA Derived from Endothelial Cells in Promoting the Formation of Colonies in Procr+ Islet Cell

Research Article

Open Access

The Role of Circular RNA Derived from Endothelial Cells in Promoting the Formation of Colonies in Procr+ Islet Cell

Yihan Deng ^1*

¹ Rutgers university

^*Corresponding author: yihandeng152@gmail.com

Published on 2 October 2025

TNS Vol.139

ISSN (Print): 2753-8826

ISSN (Online): 2753-8818

ISBN (Print): 978-1-80590-395-6

ISBN (Online): 978-1-80590-396-3

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Abstract

Diabetes mellitus is a chronic metabolic disorder characterized by impaired insulin secretion or insulin resistance, and current therapeutic approaches are unable to achieve complete remission. Recent studies have demonstrated that Procr⁺ islet cells within murine islets can form colonies and differentiate into islet organoids through co-culture with endothelial cells, subsequently reversing diabetes upon transplantation. However, the underlying mechanism by which endothelial cells promote the growth of Procr⁺ islet cells remain incompletely understood. This study aims to investigate circular RNA (circRNA) within the extracellular vesicles (EVs) of endothelial cells as a potential breakthrough. This investigation seeks to elucidate the role of circCARD6, derived from endothelial cells, in promoting the colony formation of Procr⁺ cells. I hypothesize that circCARD6 is selectively encapsulated into EVs, subsequently transferred into Procr⁺ cells, and functions as a microRNA (miRNA) sponge to modulate the expression of downstream genes, thereby enhancing cellular proliferation and differentiation. To validate this hypothesis, I employed co-culture of Procr⁺ cells and endothelial cells (ECs), followed by circRNA sequencing (RNA-seq) to identify differentially expressed circRNAs. I will employ fluorescence in situ hybridization (FISH) using BSJ-specific probes to confirm the transfer of circCARD6 from EC-derived EVs to Procr⁺ cells. Furthermore, I will use biotin-labeled miRNA pull-down assays and functional over-expression experiments to verify the sponge activity of circCARD6 on miR-31 and miR-29b-3p, as well as its impact on downstream protein expression and colony formation capacity. I expect proving that circCARD6 serves as a critical EV-derived circRNA that promotes colony formation and differentiation of Procr⁺ cells by acting as a miRNA sponge. This study provides novel insights into the mechanism of circRNA-mediated intercellular communication and identifies circCARD6 as a potential molecular target for enhancing islet regeneration. Our research findings may contribute to the development of novel therapeutic strategies for diabetes management.

Keywords:

Procr+ islet cell, CircCARD6, Endothelial cell, Extracellular vesicles

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References

[1]. Gillespie KM. Type 1 diabetes: pathogenesis and prevention. Canadian Medical Association Journal. 2006 Jun 27; 175(2): 165–70.

[2]. DeFronzo RA, Ferrannini E, Groop L, Henry RR, Herman WH, Holst JJ, et al. Type 2 Diabetes Mellitus. Nature Reviews Disease Primers. 2015 Jul 23; 1(1).

[3]. Pan FC, Wright C. Pancreas organogenesis: From bud to plexus to gland. Developmental Dynamics. 2011 Feb 17; 240(3): 530–65.

[4]. Wang D, Wang J, Bai L, Pan H, Feng H, Clevers H, et al. Long-Term Expansion of Pancreatic Islet Organoids from Resident Procr+ Progenitors. Cell. 2020 Mar; 180(6): 1198-1211.e19.

[5]. Wang J, Wang D, Chen X, Yuan S, Bai L, Liu C, et al. Isolation of mouse pancreatic islet Procr+ progenitors and long-term expansion of islet organoids in vitro. Nature Protocols [Internet]. 2022 Apr 8 [cited 2025 Jul 23]; 17(5): 1359–84.

[6]. Lammert E, Cleaver O, Melton D. Induction of Pancreatic Differentiation by Signals from Blood Vessels. Science [Internet]. 2001 Oct 19 [cited 2021 Aug 26]; 294(5542): 564–7.

[7]. Hromada C, Mühleder S, Grillari J, Redl H, Holnthoner W. Endothelial Extracellular Vesicles—Promises and Challenges. Frontiers in Physiology. 2017 May 5; 8.

[8]. Lasda E, Parker R. Circular RNAs Co-Precipitate with Extracellular Vesicles: A Possible Mechanism for circRNA Clearance. Busson P, editor. PLOS ONE. 2016 Feb 5; 11(2): e0148407.

[9]. Memczak S, Jens M, Elefsinioti A, Torti F, Krueger J, Rybak A, et al. Circular RNAs are a large class of animal RNAs with regulatory potency. Nature. 2013 Feb 27; 495(7441): 333–8.

[10]. Latreille M, Hausser J, Stützer I, Zhang Q, Hastoy B, Gargani S, et al. MicroRNA-7a regulates pancreatic β cell function. Journal of Clinical Investigation. 2014 May 1; 124(6): 2722–35.

[11]. Xu H, Guo S, Li W, Yu P. The circular RNA Cdr1as, via miR-7 and its targets, regulates insulin transcription and secretion in islet cells. Scientific Reports. 2015 Jul; 5(1).

[12]. Leïla Halidou Diallo, Mariette J, Laugero N, Touriol C, Florent Morfoisse, Prats AC, et al. Specific Circular RNA Signature of Endothelial Cells: Potential Implications in Vascular Pathophysiology. International journal of molecular sciences. 2024 Jan 4; 25(1): 680–0.

[13]. W. A. Bonner, H. R. Hulett, R. G. Sweet, L. A. Herzenberg; Fluorescence Activated Cell Sorting. Rev. Sci. Instrum. 1 March 1972; 43 (3): 404–409.

[14]. Basu S, Campbell HM, Dittel BN, Ray A. Purification of specific cell population by fluorescence activated cell sorting (FACS). J Vis Exp. 2010 Jul 10; (41): 1546.

[15]. Bayani, J. and Squire, J.A. (2004), Fluorescence In Situ Hybridization (FISH). Current Protocols in Cell Biology, 23: 22.4.1-22.4.52.

[16]. Aakash Koppula, Abdelgawad A, Jlenia Guarnerio, Batish M, Parashar V. CircFISH: A Novel Method for the Simultaneous Imaging of Linear and Circular RNAs. Cancers [Internet]. 2022 Jan 15 [cited 2024 May 5]; 14(2): 428–8.

[17]. Krützfeldt J, Poy MN, Stoffel M. Strategies to determine the biological function of microRNAs. Nature Genetics. 2006 May 30; 38(S6): S14–9.

[18]. Lewis BP, Burge CB, Bartel DP. Conserved Seed Pairing, Often Flanked by Adenosines, Indicates that Thousands of Human Genes are MicroRNA Targets. Cell. 2005 Jan; 120(1): 15–20.