Mucosally transplanted mesenchymal stem cells stimulate intestinal healing by promoting angiogenesis
Nicholas A. Manieri, Madison R. Mack, Molly D. Himmelrich, Daniel L. Worthley, Elaine M. Hanson, Lars Eckmann, Timothy C. Wang, Thaddeus S. Stappenbeck
Nicholas A. Manieri, Madison R. Mack, Molly D. Himmelrich, Daniel L. Worthley, Elaine M. Hanson, Lars Eckmann, Timothy C. Wang, Thaddeus S. Stappenbeck
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Research Article Gastroenterology

Mucosally transplanted mesenchymal stem cells stimulate intestinal healing by promoting angiogenesis

Abstract

Mesenchymal stem cell (MSC) therapy is an emerging field of regenerative medicine; however, it is often unclear how these cells mediate repair. Here, we investigated the use of MSCs in the treatment of intestinal disease and modeled abnormal repair by creating focal wounds in the colonic mucosa of prostaglandin-deficient mice. These wounds developed into ulcers that infiltrated the outer intestinal wall. We determined that penetrating ulcer formation in this model resulted from increased hypoxia and smooth muscle wall necrosis. Prostaglandin I2 (PGI2) stimulated VEGF-dependent angiogenesis to prevent penetrating ulcers. Treatment of mucosally injured WT mice with a VEGFR inhibitor resulted in the development of penetrating ulcers, further demonstrating that VEGF is critical for mucosal repair. We next used this model to address the role of transplanted colonic MSCs (cMSCs) in intestinal repair. Compared with intravenously injected cMSCs, mucosally injected cMSCs more effectively prevented the development of penetrating ulcers, as they were more efficiently recruited to colonic wounds. Importantly, mucosally injected cMSCs stimulated angiogenesis in a VEGF-dependent manner. Together, our results reveal that penetrating ulcer formation results from a reduction of local angiogenesis and targeted injection of MSCs can optimize transplantation therapy. Moreover, local MSC injection has potential for treating diseases with features of abnormal angiogenesis and repair.

Authors

Nicholas A. Manieri, Madison R. Mack, Molly D. Himmelrich, Daniel L. Worthley, Elaine M. Hanson, Lars Eckmann, Timothy C. Wang, Thaddeus S. Stappenbeck

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Figure 6

cMSCs require VEGF expression to prevent penetrating ulcers in injured Ptgir–/– mice.

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cMSCs require VEGF expression to prevent penetrating ulcers in injured P...
(A) Relative expression of Vegf mRNA in cMSCs that contain nontargeting (NT) or VEGF knockdown shRNA (two separate clones KD-1 and KD-2). P = 0.0081, 1-way ANOVA. *P < 0.05, **P < 0.01, Tukey’s post-test. n = 3 biological replicates plated at 3 sequential passages per group, repeated in technical triplicates. (B and C) Representative sections of colon (n = 5–6 wounds per group from 3 mice per group in 3 experiments) from a Ptgir–/– mouse 6 days after injury and (B) 5 days after mucosal injection of nontargeting cMSCs or (C) VEGF knockdown line KD-1 stained with anti–β-catenin antisera (green, epithelium), anti–α-SMA antisera (red, smooth muscle cells), and bisbenzimide (blue, nuclei). (D) Relative α-SMA staining in the outer muscle layer underlying wounds from Ptgir–/– mice injected with nontargeting (n = 5 wounds from 3 mice), KD-1 (n = 6 wounds from 3 mice), or KD-2 (n = 10 wounds from 4 mice) cMSCs in 3 experiments. P = 0.0265, 1-way ANOVA. *P < 0.05, Tukey’s post-test. (E) Schematic depicting how transplanted cMSCs stimulate repair. Intramucosally transplanted cMSCs can migrate to wounds in a CXCR4-dependent manner. Once cMSCs enter wounds, they provide VEGF to stimulate angiogenesis and prevent smooth muscle necrosis. Scale bar: 100 μm. Mean ± SEM.