miR-375 induces human decidua basalis-derived stromal cells to become insulin-producing cells

Abstract

This paper focuses on the development of renewable sources of isletreplacement tissue for the treatment of type I diabetes mellitus. Placental tissue-derived mesenchymal stem cells (MSCs) are a promising source for regenerative medicine due to their plasticity and easy availability. They have the potential to differentiate into insulin-producing cells. miR-375 is a micro RNA that is expressed in the pancreas and involved in islet development. Human placental decidua basalis MSCs (PDB-MSCs) were cultured from full-term human placenta. The immunophenotype of the isolated cells was checked for CD90, CD105, CD44, CD133 and CD34 markers. The MSCs (P3) were chemically transfected with hsa-miR-375. Total RNA was extracted 4 and 6 days after transfection. The expressions of insulin, NGN3, GLUT2, PAX4, PAX6, KIR6.2, NKX6.1, PDX1, and glucagon genes were evaluated using real-time qPCR. On day 6, we tested the potency of the clusters in response to the high glucose challenge and assessed the presence of insulin and NGN3 proteins via immunocytochemistry. Flow cytometry analysis confirmed that more than 90% of the cells were positive for CD90, CD105 and CD44 and negative for CD133 and CD34. Morphological changes were followed from day 2. Cell clusters formed during day 6. Insulin-producing clusters showed a deep red color with DTZ. The expression of pancreatic-specific transcription factors increased remarkably during the four days after transfection and significantly increased on day 7. The clusters were positive for insulin and NGN3 proteins, and C-peptide and insulin secretion increased in response to changes in the glucose concentration (2.8 mM and 16.7 mM). In conclusion, the MSCs could be programmed into functional insulin-producing cells by transfection of miR-375.

MicroRNAs can effectively induce formation of insulin-producing cells from mesenchymal stem cells

“…miR‐375 decreases mtpn expression and thus promotes insulin secretion by pancreatic beta cells (Poy et al , ), and maintains the normal function of these cells (Poy et al , ). miR‐375 can also induce stem cells to differentiate into pancreatic beta cells, this molecular mechanism remains poorly understood (Lahmy et al , ; Shaer et al , ). However, miR‐375 alone failed to induce the differentiation of N‐UCMSCs to IPCs in the current study.…”

MiRNAs in β-Cell Development, Identity, and Disease

“…Interestingly, acquisition of insulin expression in differentiated hESCs correlates with increased expression of miR-375 (Liao et al, 2013; Wei et al, 2013) and miR-375 overexpression is sufficient to promote pancreatic endocrine differentiation from hESCs in the absence of any extrinsic inducers (Jopling et al, 2005). In fact, overexpression of miR-375 of mesenchymal stem cells obtained from either human bone marrow or human placenta can also redirect them into functional insulin-expressing cells (Bazzini et al, 2012; Shaer et al, 2014b). These studies illustrate the crucial role of miR-375 in governing for the initiation phase of ESC differentiation and acquisition of insulin expression in β-cell precursors.…”

Genetic Engineering of Mesenchymal Stem Cells for Regenerative Medicine

“…The favourable MSC differentiation toward insulin-producing cells was achieved by the overexpression of several genes, such as betacellulin reported for BM-MSCs [257], and in the case of BM-MSCs and UC-MSCs are some data for pancreatic, and duodenal homeobox factor 1 (PDX-1) [258,259], combined with other lineage-specific genes [260,261], and pancreatic characteristic miRNAs, such as the miR-375 in the human placental decidua basalis-derived MSCs (PDB-MSCs) [262], or the insulin gene itself in hair follicle-derived MSCs (HF-MSCs) [263]. The VEGF and human IL-1 receptor antagonist (IL-1Ra)-overexpressing BM-MSCs were shown to protect islet cells from cytokines during islet transplantation [264].…”