Programming of Regulatory T Cells In Situ for Nerve Regeneration and Long-Term Patency of Vascular Grafts

Abstract

Rapid integration into the host tissue is critical for long-term patency after small diameter tissue engineering vascular grafts (sdTEVGs) transplantation. Neural recognition may be required for host integration and functionalization of the graft. However, immune rejection and inflammation hinder nerve regeneration of sdTEVGs. Here, a CRISPR/dCas9-nanocarrier was used for targeted programming of regulatory T cells (Treg cells) in situ to promote nerve regeneration of sdTEVGs by preventing excessive inflammation. Treg cells and (C-C chemokine receptor) CCR2+ macrophage recruitment occurred after transplantation. The nanodelivery system upregulated ten eleven translocation (TET2) in Treg cells in vitro. Reprogrammed Treg cells upregulated anti-inflammatory cytokines and decreased the proportion of CCR2+ macrophages. IL-6 concentrations decreased to the levels required for nerve regeneration. Implantation of CRISPR/dCas9 nanodelivery system-modified sdTEVGs in rats resulted in Treg cell editing, control of excessive inflammation, and promoted nerve regeneration. After 3 months, nerve regeneration was similar to that observed in normal blood vessels; good immune homeostasis, consistency of hemodynamics, and matrix regeneration were observed. Neural recognition promotes further integration of the graft into the host, with unobstructed blood vessels without intimal hyperplasia. Our findings provide new insights into vascular implant functionalization by the host.

In Situ‐Sprayed Bioinspired Adhesive Conductive Hydrogels for Cavernous Nerve Repair

“…IL‐6, a prototypical inflammatory factor originating from macrophages and other immune cells, has been shown to be involved in nerve regeneration. [ 33 ] Thus, we applied IL‐6 staining to examine the differences among the various treatment groups. As depicted in Figure S26 (Supporting Information), the number of IL6‐positive cells at the injury site was notably greater than that in the normal group, and there was no significant difference in the reduction in inflammation in the GAC group.…”

Applications of CRISPR Epigenome Editors in Tumor Immunology and Autoimmunity

“…94 An innovative strategy employed a Treg-specific nanodelivery system to enhance Treg potency for controlling inflammation for small diameter tissue engineering vascular grafts (sdTEVGs) transplantation. 101 This approach directly increased TET2 expression within Tregs using dCas9-VP64 and sgRNA, resulting in potent control of inflammation in vivo. 101 Utilizing the CRISPRon system in combination with nanodelivery techniques could facilitate highly specific in vivo targeting of Treg cells, focusing on regions such as the FOXP3 TSDR.…”

“…101 This approach directly increased TET2 expression within Tregs using dCas9-VP64 and sgRNA, resulting in potent control of inflammation in vivo. 101 Utilizing the CRISPRon system in combination with nanodelivery techniques could facilitate highly specific in vivo targeting of Treg cells, focusing on regions such as the FOXP3 TSDR.…”

“…Also targeting the pro-inflammatory-related genes like IFNG , TBX21 , and STAT4 with CRISPRoff might augment the function of the Treg cells since DNA methylation is also shown to be critical for the functionality of Treg cells . An innovative strategy employed a Treg-specific nanodelivery system to enhance Treg potency for controlling inflammation for small diameter tissue engineering vascular grafts (sdTEVGs) transplantation . This approach directly increased TET2 expression within Tregs using dCas9-VP64 and sgRNA, resulting in potent control of inflammation in vivo .…”

Tissue‐Penetrating Ultrasound‐Triggered Hydrogel for Promoting Microvascular Network Reconstruction

“…[ 1 ] This system is pivotal in injured tissues, where incomplete Vascular Endothelial Growth Factor (VEGF) signaling may drive organ tissue aging and damage. [ 2 ] Conversely, elevating VEGF signaling can prevent age‐related capillary loss, improving perfusion and function in various injured tissues, ultimately extending lifespan. [ 3 ] In addition, Fu et al.…”