PMID- 33726829
OWN - NLM
STAT- MEDLINE
DCOM- 20210708
LR  - 20240331
IS  - 1757-6512 (Electronic)
IS  - 1757-6512 (Linking)
VI  - 12
IP  - 1
DP  - 2021 Mar 16
TI  - Gene engineered mesenchymal stem cells: greater transgene expression and efficacy 
      with minicircle vs. plasmid DNA vectors in a mouse model of acute lung injury.
PG  - 184
LID - 10.1186/s13287-021-02245-5 [doi]
LID - 184
AB  - BACKGROUND: Acute lung injury (ALI) and in its severe form, acute respiratory 
      distress syndrome (ARDS), results in increased pulmonary vascular inflammation 
      and permeability and is a major cause of mortality in many critically ill 
      patients. Although cell-based therapies have shown promise in experimental ALI, 
      strategies are needed to enhance the potency of mesenchymal stem cells (MSCs) to 
      develop more effective treatments. Genetic modification of MSCs has been 
      demonstrated to significantly improve the therapeutic benefits of these cells; 
      however, the optimal vector for gene transfer is not clear. Given the acute 
      nature of ARDS, transient transfection is desirable to avoid off-target effects 
      of long-term transgene expression, as well as the potential adverse consequences 
      of genomic integration. METHODS: Here, we explored whether a minicircle DNA (MC) 
      vector containing human angiopoietin 1 (MC-ANGPT1) can provide a more effective 
      platform for gene-enhanced MSC therapy of ALI/ARDS. RESULTS: At 24 h after 
      transfection, nuclear-targeted electroporation using an MC-ANGPT1 vector resulted 
      in a 3.7-fold greater increase in human ANGPT1 protein in MSC conditioned media 
      compared to the use of a plasmid ANGPT1 (pANGPT1) vector (2048 +/- 567 pg/mL vs. 
      552.1 +/- 33.5 pg/mL). In the lipopolysaccharide (LPS)-induced ALI model, 
      administration of pANGPT1 transfected MSCs significantly reduced bronchoalveolar 
      lavage (BAL) neutrophil counts by 57%, while MC-ANGPT1 transfected MSCs reduced 
      it by 71% (p < 0.001) by Holm-Sidak's multiple comparison test. Moreover, 
      compared to pANGPT1, the MC-ANGPT1 transfected MSCs significantly reduced 
      pulmonary inflammation, as observed in decreased levels of proinflammatory 
      cytokines, such as tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), 
      interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), and macrophage 
      inflammatory protein-2 (MIP-2). pANGPT1-transfected MSCs significantly reduced 
      BAL albumin levels by 71%, while MC-ANGPT1-transfected MSCs reduced it by 85%. 
      CONCLUSIONS: Overall, using a minicircle vector, we demonstrated an efficient and 
      sustained expression of the ANGPT1 transgene in MSCs and enhanced the therapeutic 
      effect on the ALI model compared to plasmid. These results support the potential 
      benefits of MC-ANGPT1 gene enhancement of MSC therapy to treat ARDS.
FAU - Florian, Maria
AU  - Florian M
AD  - Ottawa Hospital Research Institute, Ottawa, ON, Canada.
FAU - Wang, Jia-Pey
AU  - Wang JP
AD  - Ottawa Hospital Research Institute, Ottawa, ON, Canada.
FAU - Deng, Yupu
AU  - Deng Y
AD  - Ottawa Hospital Research Institute, Ottawa, ON, Canada.
FAU - Souza-Moreira, Luciana
AU  - Souza-Moreira L
AD  - Ottawa Hospital Research Institute, Ottawa, ON, Canada.
FAU - Stewart, Duncan J
AU  - Stewart DJ
AD  - Ottawa Hospital Research Institute, Ottawa, ON, Canada.
AD  - The Ottawa Hospital, Ottawa, ON, Canada.
AD  - University of Ottawa, Ottawa, ON, Canada.
FAU - Mei, Shirley H J
AU  - Mei SHJ
AUID- ORCID: 0000-0003-4727-1177
AD  - Ottawa Hospital Research Institute, Ottawa, ON, Canada. smei@ohri.ca.
LA  - eng
GR  - MOP57726/CIHR/Canada
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20210316
PL  - England
TA  - Stem Cell Res Ther
JT  - Stem cell research & therapy
JID - 101527581
RN  - 0 (Lipopolysaccharides)
SB  - IM
MH  - *Acute Lung Injury/genetics/therapy
MH  - Humans
MH  - Lipopolysaccharides
MH  - Lung
MH  - *Mesenchymal Stem Cell Transplantation
MH  - *Mesenchymal Stem Cells
MH  - Mice
MH  - Transgenes
MH  - Animals
PMC - PMC7962085
OTO - NOTNLM
OT  - Acute lung injury
OT  - Acute respiratory distress syndrome
OT  - Mesenchymal stem cell
OT  - Minicircle
COIS- The funding institution had no role in the conception, design or conduct of the 
      study, data collection or analysis, interpretation or presentation of the data, 
      or preparation, review, or approval of the manuscript. We would also like to 
      declare the following conflicts of interest: DJS holds a patent for MSC therapy 
      for the treatment of acute lung injury and SHJM and YD have received personal 
      fees from Northern Therapeutics Inc. that are outside of this submitted work. The 
      remaining authors have disclosed that they do not have any conflicts of interest.
EDAT- 2021/03/18 06:00
MHDA- 2021/07/09 06:00
PMCR- 2021/03/16
CRDT- 2021/03/17 05:48
PHST- 2020/10/13 00:00 [received]
PHST- 2021/02/25 00:00 [accepted]
PHST- 2021/03/17 05:48 [entrez]
PHST- 2021/03/18 06:00 [pubmed]
PHST- 2021/07/09 06:00 [medline]
PHST- 2021/03/16 00:00 [pmc-release]
AID - 10.1186/s13287-021-02245-5 [pii]
AID - 2245 [pii]
AID - 10.1186/s13287-021-02245-5 [doi]
PST - epublish
SO  - Stem Cell Res Ther. 2021 Mar 16;12(1):184. doi: 10.1186/s13287-021-02245-5.