Transplantation of CRISPRa system engineered IL10-overexpressing bone marrow-derived mesenchymal stem cells for the treatment of myocardial infarction in diabetic mice

Abstract

Background Myocardial infarction (MI) is a common cause of mortality in people. Mesenchymal stem cell (MSC) has been shown to exert therapeutic potential to treat myocardial infarction (MI). However, in patients with diabetes, the diabetic environment affected MSCs activity and could impair the efficacy of treatment. Interleukin-10 (IL-10) has been shown to attenuate MI by suppressing inflammation. In current study, the combination of MSC transplantation with IL-10 was evaluated in a diabetic mice model with MI. Methods We engineered bone marrow derived MSCs (BM-MSCs) to overexpress IL-10 by using CRISPR activation. We established the diabetic mice model with MI and monitored the IL-10 expression after BM-MSCs transplantation. We also evaluated the effects of BM-MSCs transplantation on inflammatory response, cell apoptosis, cardiac function and angiogenesis. Results CRISPR activation system enabled overexpression of IL-10 in BM-MSCs. Transplantation of BM-MSCs overexpressing IL-10 resulted in IL-10 expression in heart after transplantation. Transplantation of BM-MSCs overexpressing IL-10 inhibited inflammatory cell infiltration and pro-inflammatory cytokines production, improved cardiac functional recovery, alleviated cardiac injury, decreased apoptosis of cardiac cells and increased angiogenesis. Conclusion In summary, we have demonstrated the therapeutic potential of IL-10 overexpressed BM-MSCs in the treatment of MI in diabetic mice. Electronic supplementary material The online version of this article (10.1186/s13036-019-0163-6) contains supplementary material, which is available to authorized users.

Functional heterogeneity of mesenchymal stem cells from natural niches to culture conditions: implications for further clinical uses

“…Thereby, using CRISPR-Cas9 knockout and transcriptional activation systems, we were able to create both PAI-1 knockout and PAI-1 over-expressing human endometrium-derived mesenchymal stem cells (hMESCs), respectively [241]. It was also reported the use of engineered BM-MSC overexpressing IL-10 using CRISPR activation to treated myocardial infarction in diabetic mice [242].…”

“…Prolonged survival in a prostate cancer lung metastasis model, compared to controls [ 233 , 234 ] IFN-γ hMSC engineered hBM-MSC In vitro Inhibition of proliferation and induction of apoptosis in leukemia cells [ 235 ] Ad-FKN engineered adenoviral vector fractalkine gene In vitro In vivo * Ad-fractalkine mediates antitumor effects by induction of both innate and adaptive immunity [ 236 ] IL-12 expressed hBM-MSC In vitro In vivo * Prevention of breast cancer metastasis into the lymph nodes and internal organs as well as increased tumor cell apoptosis and an antiangiogenic effect on tumor stroma [ 237 ] (CRISPR)/Cas9 hMESCs BM-MSC In vitro In vitro In vivo Obtain PAI-1 knockout and PAI-1 overexpressing hMESCs, provides evidence of successful and effective MSCs secretome managing via CRISPR/Cas9 genome editing technology Overexpression of IL-10 in BM-MSCs. Transplantation of BM-MSCs overexpressing IL-10 inhibited inflammatory cell infiltration and pro-inflammatory cytokines production, improved cardiac functional recovery, alleviated cardiac injury, decreased apoptosis of cardiac cells and increased angiogenesis [ 241 ] [ 242 ] * animal model …”

Genetic Engineering as a Strategy to Improve the Therapeutic Efficacy of Mesenchymal Stem/Stromal Cells in Regenerative Medicine

“…Chagas disease cardiomyopathy Silva et al, 2018a Mouse bone marrow Adenovirus Cirrhosis Fiore et al, 2015 Integrin linked kinase (ILK) Mini-pig bone marrow Adenovirus Myocardial infarction Mao Q. et al, 2014 Rats bone marrow Adenovirus Myocardial infarction Mao et al, 2013 Integrin the most used in gene therapy, whether in preclinical or clinical studies, as they guarantee high process efficiency, as shown by the increased number of phase I clinical studies evaluating the safety of gene therapies using lentiviruses conducted in recent years (Milone and O'Doherty, 2018). Recently, new genetic modification tools arose in order to promote insertion, deletion or correction of genes at specific sites in the genome, and MSCs have been used as a target for these new modifying tools for applications in different diseases (Torres et al, 2014;van den Akker et al, 2016;Gerace et al, 2017;Li et al, 2018;Meng et al, 2019). The sitespecific integration of genes can be achieved using tools such as Zinc Finger Nuclease (ZFN), Transcription Activator-Like Effector Nuclease (TALENS) or Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR/Cas9), which are nucleases capable of recognizing and direct the integration of genes in a site-specific manner.…”

“…Furthermore, anti-inflammatory interleukins and regulators of oxidative factors were explored in the production of genetically engineered MSCs. IL-10 overexpression improved the therapeutic effects of MSCs by reducing by 2.5-fold the production of proinflammatory cytokines (TNF and IL-1β) and promoting a 3-fold reduction of the heart infarct size when compared to control MSCs (Meng et al, 2017(Meng et al, , 2019. In addition, injection of MSCs modified to overexpress IL-33, a cytokine known to induce Th2 and Treg responses (Schmitz et al, 2005) resulted in a significant improvement of heart function and reduced inflammation compared to vector control MSCs in rats with myocardial infarction (Chen et al, 2019).…”

“…Recently, new genetic modification tools arose in order to promote insertion, deletion or correction of genes at specific sites in the genome, and MSCs have been used as a target for these new modifying tools for applications in different diseases ( Torres et al, 2014 ; van den Akker et al, 2016 ; Gerace et al, 2017 ; Li et al, 2018 ; Meng et al, 2019 ). The site-specific integration of genes can be achieved using tools such as Zinc Finger Nuclease (ZFN), Transcription Activator-Like Effector Nuclease (TALENS) or Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR/Cas9), which are nucleases capable of recognizing and direct the integration of genes in a site-specific manner.…”