sTNFRII-Fc modification protects human UC-MSCs against apoptosis/autophagy induced by TNF-α and enhances their efficacy in alleviating inflammatory arthritis

Abstract

Background Tumor necrosis factor (TNF)-α inhibitors represented by Etanercept (a fusion protein containing soluble TNF receptor II (sTNFRII) and the Fc segment of human IgG1) play a pivotal role in Rheumatoid arthritis (RA) treatment. However, long-term use increases the risk of infection and tumors for their systemic inhibition of TNF-α, which disrupts the regular physiological function of this molecular. Mesenchymal stem cells (MSCs)-based delivery system provides new options for RA treatment with their “homing” and immune-regulation capacities, whereas inflammatory environment (especially TNF-α) is not conducive to MSCs' therapeutic effects by inducing apoptosis/autophagy. Here, we constructed a strain of sTNFRII-Fc-expressing MSCs (sTNFRII-MSC), aiming to offset the deficiency of those two interventions. Methods Constructed sTNFRII-Fc lentiviral vector was used to infect human umbilical cord-derived MSCs, and sTNFRII-MSC stable cell line was generated by monoclonal cultivation. In vitro and vivo characteristics of sTNFRII-MSC were assessed by coculture assay and an acute inflammatory model in NOD/SCID mice. The sTNFRII-MSC were transplanted into CIA model, pathological and immunological indicators were detected to evaluate the therapeutic effects of sTNFRII-MSC. The distribution of sTNFRII-MSC was determined by immunofluorescence assay. Apoptosis and autophagy were analyzed by flow cytometry, western blot and immunofluorescence. Results sTNFRII-Fc secreted by sTNFRII-MSC present biological activity both in vitro and vivo. sTNFRII-MSC transplantation effectively alleviates mice collagen-induced arthritis (CIA) via migrating to affected area, protecting articular cartilage destruction, modulating immune balance and sTNFRII-MSC showed prolonged internal retention via resisting apoptosis/autophagy induced by TNF-α. Conclusion sTNFRII-Fc modification protects MSCs against apoptosis/autophagy induced by TNF-α, in addition to releasing sTNFRII-Fc neutralizing TNF-α to block relevant immune-inflammation cascade, and thus exert better therapeutic effects in alleviating inflammatory arthritis.

Activation of KYN-AHR axis impairs the chondrogenic and chondroprotective effects of human umbilical cord-derived mesenchymal stem cells on osteoarthritis

“…MSCs can differentiate into chondrocytes and induce the resident progenitor cells differentiation, and secret lots of cytokines to promote cartilage regeneration [38,39]. However, the chondrogenic differentiation of MSCs in vivo is instable and susceptible to the microenvironment [15,34]. The component of OA SF is complicated and may have a potential effect on chondrogenesis of MSCs, there study have showed the catabolic factors such as IL-1 or TNF-α in OA SF inhibit chondrogenesis of MSCs, but blocking IL-1 or TNF-α just partially overcame the inhibitory effect [40], which indicates that there are additional factors present in OA SF that participate in inhibition of MSCs chondrogenic differentiation.…”

“…Brie y, chondrocyte spheroids were treated with 8 M guanidine HCL containing 0.05 M acetate, pH 5.8 and proteinase inhibitors overnight. Then, the absorbance of eluent was measured at 600 nm using a microplate reader as previously described [15].…”

“…Some evidence suggests that the effect of MSCs therapeutics will be affected by the in ammatory environment. There is report found that the in ammatory microenvironment of acute myocardial infarction has an inhibitory effect on the stem cells potential for regenerating the injured myocardium and secretion of critical cytokines with pro-in ammatory properties cause the cellular death and hinder the stem cells proliferation and differentiation [15]. In addition, Lou found that metabolic abnormalities and the in ammatory milieu affect the differentiation potential of adipose-derived mesenchymal stem cells, impairing their functions in maintaining tissue repair and homeostasis [16].…”

Apoptotic cell death in disease—Current understanding of the NCCD 2023

“…Mice with defects in FASL or TNF signaling are protected against arthritis induced by immunization with xenogeneic type II collagen in complete Freund's adjuvant [911][912][913][914]. Similar protection was observed in mice transplanted with mesenchymal stem cells engineered to express TNF inhibitors [915]. In keeping with this evidence, the myeloid cell specific deletion of Fas or the administration of antibodies that target both TNF and chemokine (C-X-C motif) ligand 10 (CXCL10) resulted in accelerated disease resolution in a model of rheumatoid arthritis induced by K/BxN serum transfer [916,917].…”

The role of TNF-α in the fate regulation and functional reprogramming of mesenchymal stem cells in an inflammatory microenvironment

“…Similarly, 20 ng/ml TNF-α stimulation of human umbilical cord MSCs for 24 h increased the expression of autophagy-related protein microtubule-associated protein 1 light chain B-II (LC3B-II) and lowered the expression of autophagy inhibitory molecule tribbles homology protein 3 (TRIB3) ( 15 ). Therefore, in the inflammatory microenvironment, and especially during in vitro experiments, the presence of high concentrations of inflammatory factors promotes the survival of MSCs via autophagy.…”