Knockout of integrin β1 in induced pluripotent stem cells accelerates skin-wound healing by promoting cell migration in extracellular matrix

Abstract

Background Induced pluripotent stem cells (iPSCs) have the potential to promote wound healing; however, their adhesion to the extracellular matrix (ECM) might decrease iPSC migration, thereby limiting their therapeutic potential. Integrin β1 (Itgb1) is the major integrin subunit that mediates iPSC-ECM adhesion, suggesting that knocking out Itgb1 might be an effective method for enhancing the therapeutic efficacy of iPSCs. Methods We knocked out Itgb1 in mouse iPSCs and evaluated its effects on the therapeutic potential of topically applied iPSCs, as well as their underlying in vivo and in vitro mechanisms. Results The Itgb1-knockout (Itgb1-KO) did not change iPSC pluripotency, function, or survival in the absence of embedding in an ECM gel but did accelerate wound healing, angiogenesis, blood perfusion, and survival in skin-wound lesions. However, embedding in an ECM gel inhibited the in vivo effects of wild-type iPSCs but not those of Itgb1-knockout iPSCs. Additionally, in vitro results showed that Itgb1-knockout decreased iPSC-ECM adhesion while increasing ECM-crossing migration. Moreover, ECM coating on the culture surface did not change cell survival, regardless of Itgb1 status; however, the in vivo and in vitro functions of both Itgb1-knockout and wild-type iPSCs were not affected by the presence of agarose gel, which does not contain integrin-binding sites. Knockout of Integrin α4 (Itga4) did not change the above-mentioned cellular and therapeutic functions of iPSCs. Conclusions Itgb1-knockout increased iPSCs migration and the wound-healing-promoting effect of topically applied iPSCs. These findings suggest the inhibition of Itgb1 expression is a possible strategy for increasing the efficacy of iPSC therapies.

Wnt4 increases the thickness of the epidermis in burn wounds by activating canonical Wnt signalling and decreasing the cell junctions between epidermal cells

“…ITGα6 was reported to be a positive factor for wound healing [ 40 , 41 ]. The role of ITGβ1 in wound healing is controversial [ 42 , 43 ]. Burn wound healing is a complex process and not all the results are consistent with the literature.…”

Mesenchymal stem cells in craniofacial reconstruction: a comprehensive review

“…To address this issue, both reducing adhesion and enhancing migratory capacity are viable solutions. If the Itgb1 gene in iPSCs is knocked out to reduce adhesion to the ECM, wound healing can be accelerated, and the survival rate of transplanted skin can be improved ( Ren et al, 2022 ). On the other hand, some proteins such as heat shock protein-90α (Hsp-90α) can activate the Akt signaling pathway, promoting the migration of iPSCs-derived keratinocytes towards the edges of burn wounds ( Wu et al, 2019 ).…”