Preservation of reserve intestinal epithelial stem cells following severe ischemic injury

Abstract

Intestinal ischemia is an abdominal emergency with a mortality rate >50%, leading to epithelial barrier loss and subsequent sepsis. Epithelial renewal and repair after injury depend on intestinal epithelial stem cells (ISC) that reside within the crypts of Lieberkühn. Two ISC populations critical to epithelial repair have been described: 1) active ISC (aISC; highly proliferative; leucine-rich-repeat-containing G protein-coupled receptor 5 positive, sex determining region Y-box 9 positive) and 2) reserve ISC [rISC; less proliferative; homeodomain only protein X (Hopx)+]. Yorkshire crossbred pigs (8–10 wk old) were subjected to 1–4 h of ischemia and 1 h of reperfusion or recovery by reversible mesenteric vascular occlusion. This study was designed to evaluate whether ISC-expressing biomarkers of aISCs or rISCs show differential resistance to ischemic injury and different contributions to the subsequent repair and regenerative responses. Our data demonstrate that, following 3–4 h ischemic injury, aISC undergo apoptosis, whereas rISC are preserved. Furthermore, these rISC are retained ex vivo in spheroids in which cell populations are enriched in the rISC biomarker Hopx. These cells appear to go on to provide a proliferative pool of cells during the recovery period. Taken together, these data indicate that Hopx+ cells are resistant to injury and are the likely source of epithelial renewal following prolonged ischemic injury. It is therefore possible that targeting reserve stem cells will lead to new therapies for patients with severe intestinal injury. NEW & NOTEWORTHY The population of reserve less-proliferative intestinal epithelial stem cells appears resistant to injury despite severe epithelial cell loss, including that of the active stem cell population, which results from prolonged mesenteric ischemia. These cells can change to an activated state and are likely indispensable to regenerative processes. Reserve stem cell targeted therapies may improve treatment and outcome of patients with ischemic disease.

Stem Cell Therapy for Intestinal Ischemia-Reperfusion: Current State and Perspectives

“…towards the crypt base, with an architectural preservation of the crypts [7]. These findings stand in line with the theory that following tissue insult the potent regeneration area is preserved, as the crypts are assumed to inhabit the ISCs.…”

Intestinal Mucosal Wound Healing and Barrier Integrity in IBD–Crosstalk and Trafficking of Cellular Players

“…It is also worth mentioning, that there are two distinct ISC populations: Crypt base columnar (CBC) cells, which are actively proliferating and reserve intestinal stem cells (rISC) that are quiescent stem cells until activated upon injury. In line with this, Gonzalez et al (37) showed that Hopx + cells (rISC) are resistant to injury and are the likely source of epithelial renewal following prolonged ischemic injury (37).…”

Tackling Ischemic Reperfusion Injury With the Aid of Stem Cells and Tissue Engineering

“…In addition, by modulating spheroid size, oxygen availability (and thus, normoxia/hypoxia) in the spheroid core can be modulated to a certain extent. Such platforms have been used to study the mechanism of IRI, the role of SCs in the regeneration process, and to produce biologics with potential therapies in renal (Xu et al, 2016;, hepatic (Sun et al, 2018;Olander et al, 2019), intestinal (Gonzalez et al, 2019), cardiovascular (Chen et al, 2007;Suzuki et al, 2019;Sfriso and Rieben, 2020), lung (Pagano et al, 2017), skin (Park et al, 2017), fat (Mineda et al, 2015), and neural (Davenport Jones et al, 1998;Serganova et al, 2004;Fu et al, 2015;Liu et al, 2015;…”

“…To study intestinal IRI, porcine intestinal epithelial stem cells (ISCs) were harvested after a prolonged duration of vascular occlusion to guarantee deep epithelial loss. Afterward, spheroid cultures were established to determine the impact of different durations IRI (1-4 h) on ISCs in vitro and in the ischemic crypt and the contribution of these cells to mucosal repair (Gonzalez et al, 2019). This study highlighted a closely related behavior of ISCs in both 3D in vitro and in vivo platforms based on the expression profile of the genes leucine-rich repeat-containing G protein-coupled receptor 5 (Lgr5), SOX9 Lrig HOPX.…”