TY - JOUR
T1 - Extracellular vesicle transfer of lncRNA H19 splice variants to cardiac cells
AU - Vilaça, Andreia
AU - Jesus, Carlos
AU - Lino, Miguel
AU - Hayman, Danika
AU - Emanueli, Costanza
AU - Terracciano, Cesare M.
AU - Fernandes, Hugo
AU - de Windt, Leon J.
AU - Ferreira, Lino
N1 - Funding Information:
The authors would like to acknowledge Crioestaminal ( www.crioestaminal.pt ) for providing samples and other helpful information and Dr. Yingqun Huang for providing the lncRNA H19 plasmid. The authors would further like to acknowledge funding by the FCT PhD Studentships ( SFRH/BD/119187/2016 and SFRH/BD/144092/2019 ), Programa Operacional Competividade e Internacionaliza\u00E7\u00E3o ( POCI ) na sua componente FEDER e pelo or\u00E7amento da Funda\u00E7\u00E3o para a Ci\u00EAncia e a Tecnologia na sua componente OE (Project 2022.03308.PTDC ), EC projects RESET aging (ref. 952266 ) and REBORN (ref. 101091852 ), and PRR project HfPT-Health from Portugal (ref. 02/C05-i01.01/2022.PC644937233-00000047 ). L.J.d.W. acknowledges support from the Dutch Cardiovascular Alliance ( ARENA-PRIME ). L.J.d.W. was further supported by a VICI award ( 918-156-47 ) from the Dutch Research Council , Marie Sklodowska-Curie grant agreement 813716 (TRAIN-HEART), and a PPP allowance made available by Health\u223CHolland , Top Sector Life Sciences & Health, under agreement LSHM21068 to stimulate public-private partnerships .
Funding Information:
The authors would like to acknowledge Crioestaminal (www.crioestaminal.pt) for providing samples and other helpful information and Dr. Yingqun Huang for providing the lncRNA H19 plasmid. The authors would further like to acknowledge funding by the FCT PhD Studentships (SFRH/BD/119187/2016 and SFRH/BD/144092/2019), Programa Operacional Competividade e Internacionaliza\u00E7\u00E3o (POCI) na sua componente FEDER e pelo or\u00E7amento da Funda\u00E7\u00E3o para a Ci\u00EAncia e a Tecnologia na sua componente OE (Project 2022.03308.PTDC), EC projects RESETaging (ref. 952266) and REBORN (ref. 101091852), and PRR project HfPT-Health from Portugal (ref. 02/C05-i01.01/2022.PC644937233-00000047). L.J.d.W. acknowledges support from the Dutch Cardiovascular Alliance (ARENA-PRIME). L.J.d.W. was further supported by a VICI award (918-156-47) from the Dutch Research Council, Marie Sklodowska-Curie grant agreement 813716 (TRAIN-HEART), and a PPP allowance made available by Health\u223CHolland, Top Sector Life Sciences & Health, under agreement LSHM21068 to stimulate public-private partnerships. A.V. C.J. M.L. and D.H. conducted the experiments. A.V. C.J. H.F. L.J.d.W. and L.F. designed the experiments. A.V. C.J. M.L. D.H. C.E. C.M.T. H.F. L.J.d.W. and L.F. analyzed the experiments. A.V. and C.J. analyzed the data. A.V. C.J. H.F. L.J.d.W. and L.F. wrote the paper. The authors declare no competing interests.
Publisher Copyright:
© 2024 The Author(s)
PY - 2024/9/10
Y1 - 2024/9/10
N2 - The delivery of therapeutic long non-coding RNAs (lncRNA) to the heart by extracellular vesicles (EVs) is promising for heart repair. H19, a lncRNA acting as a major regulator of gene expression within the cardiovascular system, is alternatively spliced, but the loading of its different splice variants into EVs and their subsequent uptake by recipient cardiac cells remain elusive. Here, we dissected the cellular expression of H19 splice variants and their loading into EVs secreted by Wharton-Jelly mesenchymal stromal/stem cells (WJ-MSCs). We demonstrated that overexpression of the mouse H19 gene in WJ-MSCs induces the expression of H19 splice variants at different levels. Interestingly, EVs isolated from the H19-transfected WJ-MSCs (EV-H19) showed similar expression levels for all tested splice variant sets. In vitro, we further demonstrated that EV-H19 was taken up by cardiomyocytes, fibroblasts, and endothelial cells (ECs). Finally, analysis of EV tropism in living rat myocardial slices indicated that EVs were internalized mostly by cardiomyocytes and ECs. Collectively, our results indicated that EVs can be loaded with different lncRNA splice variants and successfully internalized by cardiac cells.
AB - The delivery of therapeutic long non-coding RNAs (lncRNA) to the heart by extracellular vesicles (EVs) is promising for heart repair. H19, a lncRNA acting as a major regulator of gene expression within the cardiovascular system, is alternatively spliced, but the loading of its different splice variants into EVs and their subsequent uptake by recipient cardiac cells remain elusive. Here, we dissected the cellular expression of H19 splice variants and their loading into EVs secreted by Wharton-Jelly mesenchymal stromal/stem cells (WJ-MSCs). We demonstrated that overexpression of the mouse H19 gene in WJ-MSCs induces the expression of H19 splice variants at different levels. Interestingly, EVs isolated from the H19-transfected WJ-MSCs (EV-H19) showed similar expression levels for all tested splice variant sets. In vitro, we further demonstrated that EV-H19 was taken up by cardiomyocytes, fibroblasts, and endothelial cells (ECs). Finally, analysis of EV tropism in living rat myocardial slices indicated that EVs were internalized mostly by cardiomyocytes and ECs. Collectively, our results indicated that EVs can be loaded with different lncRNA splice variants and successfully internalized by cardiac cells.
KW - extracellular vesicles
KW - H19 lncRNA
KW - MT: Delivery Strategies
KW - RNA therapeutics
KW - splice variants
U2 - 10.1016/j.omtn.2024.102233
DO - 10.1016/j.omtn.2024.102233
M3 - Article
SN - 2162-2531
VL - 35
JO - Molecular Therapy - Nucleic Acids
JF - Molecular Therapy - Nucleic Acids
IS - 3
M1 - 102233
ER -