Reversible secretome and signaling defects in diabetic mesenchymal stem cells from peripheral arterial disease patients

• Type of Material: Article
• Authors: Tatiana Chadid, Emory University; Andrew Morris, Emory University; Alexandra Surowiec, Vanderbilt University; Scott Robinson, University of Michigan; Maiko Sasaki, Emory University; Jacques Galipeau, Emory University; Brian Pollack, Emory University; Luke Brewster, Emory University
• Language: English
• Date: 2018-12-01
• Publisher: MOSBY-ELSEVIER
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2018 Society for Vascular Surgery
• License: Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1016/j.jvs.2018.05.223
• Title of Journal or Parent Work: JOURNAL OF VASCULAR SURGERY
• Volume: 68
• Issue: 6
• Start Page: 137S
• End Page: +
• Grant/Funding Information: American Heart Association IRG 13IRG14740001 (LPB), 1KO8HL119592 (LPB); PHS Grant UL1TR000454.
• Keywords: Life Sciences & Biomedicine; Peripheral Vascular Disease; INFLAMMATION; REVASCULARIZATION; ANGIOGENESIS; CRITICAL LIMB ISCHEMIA; STROMAL CELLS; Cardiovascular System & Cardiology; Type 2; THERAPY; Science & Technology; TRANSPLANTATION; MARROW MONONUCLEAR-CELLS; Cellular therapy; Mesenchymal stem cells; Surgery; Angiogenesis; Growth factors; BONE-MARROW; NEOVASCULARIZATION CAPACITY; Peripheral arterial disease; Diabetes mellitus; Epidermal growth factor
• Research Categories: Biology, Cell; Health Sciences, Rehabilitation and Therapy; Health Sciences, Epidemiology; Health Sciences, Medicine and Surgery; Biology, Neuroscience

Abstract

Objective: Regenerative medicine seeks to stall or to reverse the pathologic consequences of chronic diseases. Many people with diabetes have peripheral arterial disease (PAD), which increases their already high risk of major amputation. Cellular therapies are a promising regenerative medicine approach to PAD that can be used to focally inject regenerative cells to endangered tissue beds. Mesenchymal stem cells (MSCs) are known to promote tissue regeneration through stromal support and paracrine stimulation of new blood vessels (angiogenesis). Whereas little is known about human diabetic MSCs (dMSCs), particularly those from patients with PAD, dMSCs have a limited expansion capacity but can be improved with human platelet lysate (PL) supplementation. PL is rich in many growth factors, including epidermal growth factor (EGF), which is known to be important to cell proliferation and survival signaling pathways. We hypothesize that dMSCs have a reversible defect in EGF receptor pathways. The objective of this work was to test this hypothesis using dMSCs from PAD patients. Methods: The secretome expression of EGF and prominent angiogens was characterized from bone marrow (BM)-derived and adipose tissue-derived (ATD) dMSCs from five patients (six limbs) undergoing major amputation. Western blot was used to characterize the AKT and extracellular signal-regulated protein kinases 1 and 2 expression in dMSCs under standard culture (5% fetal bovine serum plus fibroblast growth factor 2 [FGF2]), 5% human PL, or 5% fetal bovine serum plus EGF. Healthy donor MSCs were control cells. The angiogenic activity of BM- and ATD-dMSCs was tested on human umbilical vein endothelial cells (ECs). Paired t-test, analysis of variance, and Kruskal-Wallis tests were used as appropriate. Results: Both BM- and ATD-dMSCs had typical MSC surface marker expression and similar expansion profiles, and they did not express EGF in their secretome. PL supplementation of dMSCs improved AKT signaling, but they were resistant to FGF2 activation of extracellular signal-regulated protein kinases 1 and 2. EGF supplementation led to similar AKT expression as with PL, but PL had greater phosphorylation of AKT at 30 and 60 minutes. The conditioned media from both BM- and ATD-dMSCs had robust levels of prominent angiogens (vascular endothelial growth factor, monocyte chemoattractant protein 1, hepatocyte growth factor), which stimulated EC proliferation and migration, and the co-culture of dMSCs with ECs led to significantly longer EC sprouts in three-dimensional gel than EC-alone pellets. Conclusions: PL and EGF supplementation improves AKT expression in dMSCs over that of FGF2, but PL improved pAKT over that of EGF. Thus, PL supplementation strategies may improve AKT signaling, which could be important to MSC survival in cellular therapies. Furthermore, BM- and ATD-dMSCs have similar secretomes and robust in vitro angiogenic activity, which supports pursuing dMSCs from both reservoirs in regenerative medicine strategies. Clinical Relevance: Cellular therapies with mesenchymal stem cells (MSCs) hold great promise in the treatment of chronic diseases like peripheral arterial disease (PAD). Diabetic patients have a premature onset and rapid progression of PAD, but the suitability of MSCs in diabetic patients is not clear. We have found that both bone marrow and adipose tissue-derived MSCs had robust angiogenic effects in vitro. The signaling defects identified in the epidermal growth factor signaling axis were partially recovered with platelet lysate supplementation. By correcting the defects identified, we may be able to improve the cell survival and expansion of MSCs for use in diabetic patients with PAD.