Chromatographic Scalable Method to Isolate Engineered Extracellular Vesicles Derived from Mesenchymal Stem Cells for the Treatment of Liver Fibrosis in Mice

被引：3
作者：
Dominguez, Luciana M. M. [1 ]
Bueloni, Barbara [1 ]
Cantero, Ma. Jose [1 ]
Albornoz, Milagros [1 ]
Pacienza, Natalia [2 ]
Biani, Celeste [3 ]
Luzzani, Carlos [3 ]
Miriuka, Santiago [3 ]
Garcia, Mariana [1 ]
Atorrasagasti, Catalina [1 ]
Yannarelli, Gustavo [2 ]
Bayo, Juan [1 ]
Fiore, Esteban [1 ]
Mazzolini, Guillermo [1 ,4 ]
机构：
[1] Univ Austral, Lab Terapia Genica, Inst Invest Med Traslac IIMT, CONICET, RA-1629 Buenos Aires, DF, Argentina
[2] Univ Favaloro, Inst Med Traslac Trasplante & Bioingn IMeTTyB, CONICET, RA-1078 Buenos Aires, Argentina
[3] Fleni, LIAN, CONICET, RA-1625 Buenos Aires, DF, Argentina
[4] Univ Austral, Hosp Univ Austral, Liver Unit, CONICET, RA-1629 Buenos Aires, DF, Argentina
来源：
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES | 2023年 / 24卷 / 11期
关键词：
liver fibrosis; extracellular vesicles; mesenchymal stromal cells; chromatography EVs isolation; engineered EVs; STROMAL CELLS; EXOSOMES;
D O I：
10.3390/ijms24119586
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
New therapeutic options for liver cirrhosis are needed. Mesenchymal stem cell (MSC)-derived extracellular vesicles (EVs) have emerged as a promising tool for delivering therapeutic factors in regenerative medicine. Our aim is to establish a new therapeutic tool that employs EVs derived from MSCs to deliver therapeutic factors for liver fibrosis. EVs were isolated from supernatants of adipose tissue MSCs, induced-pluripotent-stem-cell-derived MSCs, and umbilical cord perivascular cells (HUCPVC-EVs) by ion exchange chromatography (IEC). To produce engineered EVs, HUCPVCs were transduced with adenoviruses that code for insulin-like growth factor 1 (AdhIGF-I-HUCPVC-EVs) or green fluorescent protein. EVs were characterized by electron microscopy, flow cytometry, ELISA, and proteomic analysis. We evaluated EVs' antifibrotic effect in thioacetamide-induced liver fibrosis in mice and on hepatic stellate cells in vitro. We found that IEC-isolated HUCPVC-EVs have an analogous phenotype and antifibrotic activity to those isolated by ultracentrifugation. EVs derived from the three MSCs sources showed a similar phenotype and antifibrotic potential. EVs derived from AdhIGF-I-HUCPVC carried IGF-1 and showed a higher therapeutic effect in vitro and in vivo. Remarkably, proteomic analysis revealed that HUCPVC-EVs carry key proteins involved in their antifibrotic process. This scalable MSC-derived EV manufacturing strategy is a promising therapeutic tool for liver fibrosis.
引用
收藏
页数：19
相关论文
共 55 条
[1]   Strategies for scalable manufacturing and translation of MSC-derived extracellular vesicles [J].
Adlerz, Katrina ;
Patel, Divya ;
Rowley, Jon ;
Ng, Kelvin ;
Ahsan, Tabassum .
STEM CELL RESEARCH, 2020, 48
[2]   Characterization of extracellular vesicles and synthetic nanoparticles with four orthogonal single-particle analysis platforms [J].
Arab, Tanina ;
Mallick, Emily R. ;
Huang, Yiyao ;
Dong, Liang ;
Liao, Zhaohao ;
Zhao, Zezhou ;
Gololobova, Olesia ;
Smith, Barbara ;
Haughey, Norman J. ;
Pienta, Kenneth J. ;
Slusher, Barbara S. ;
Tarwater, Patrick M. ;
Tosar, Juan Pablo ;
Zivkovic, Angela M. ;
Vreeland, Wyatt N. ;
Paulaitis, Michael E. ;
Witwer, Kenneth W. .
JOURNAL OF EXTRACELLULAR VESICLES, 2021, 10 (06)
[3]   Re-Engineering Extracellular Vesicles as Smart Nanoscale Therapeutics [J].
Armstrong, James P. K. ;
Holme, Margaret N. ;
Stevens, Molly M. .
ACS NANO, 2017, 11 (01) :69-83
[4]   Clinical requirements for extracellular vesicle assays [J].
Ayers, Lisa ;
Pink, Ryan ;
Carter, David Raul Francisco ;
Nieuwland, Rienk .
JOURNAL OF EXTRACELLULAR VESICLES, 2019, 8 (01)
[5]   Isolation of Exosomes from Blood Plasma: Qualitative and Quantitative Comparison of Ultracentrifugation and Size Exclusion Chromatography Methods [J].
Baranyai, Tamas ;
Herczeg, Kata ;
Onodi, Zsofia ;
Voszka, Istvan ;
Modos, Karoly ;
Marton, Nikolett ;
Nagy, Gyoergy ;
Maeger, Imre ;
Wood, Matthew J. ;
El Andaloussi, Samir ;
Palinkas, Zoltan ;
Kumar, Vikas ;
Nagy, Pater ;
Kittel, Agnes ;
Buzas, Edit Iren ;
Ferdinandy, Peter ;
Giricz, Zoltan .
PLOS ONE, 2015, 10 (12)
[6]   Liver fibrosis [J].
Bataller, R ;
Brenner, DA .
JOURNAL OF CLINICAL INVESTIGATION, 2005, 115 (02) :209-218
[7]   Bioinformatic analysis of RHO family of GTPases identifies RAC1 pharmacological inhibition as a new therapeutic strategy for hepatocellular carcinoma [J].
Bayo, Juan ;
Fiore, Esteban J. ;
Maria Dominguez, Luciana ;
Jose Cantero, Maria ;
Ciarlantini, Matias S. ;
Malvicini, Mariana ;
Atorrasagasti, Catalina ;
Gabriela Garcia, Mariana ;
Rossi, Mario ;
Cavasotto, Claudio ;
Martinez, Elisabeth ;
Comin, Julieta ;
Mazzolini, Guillermo D. .
GUT, 2021, 70 (07) :1362-1374
[8]   Human Umbilical Cord Perivascular Cells Exhibited Enhanced Migration Capacity towards Hepatocellular Carcinoma in Comparison with Bone Marrow Mesenchymal Stromal Cells: A Role for Autocrine Motility Factor Receptor [J].
Bayo, Juan ;
Fiore, Esteban ;
Aquino, Jorge B. ;
Malvicini, Mariana ;
Rizzo, Manglio ;
Peixoto, Estanislao ;
Alaniz, Laura ;
Piccioni, Flavia ;
Bolontrade, Marcela ;
Podhajcer, Osvaldo ;
Garcia, Mariana G. ;
Mazzolini, Guillermo .
BIOMED RESEARCH INTERNATIONAL, 2014, 2014
[9]   Increased Migration of Human Mesenchymal Stromal Cells by Autocrine Motility Factor (AMF) Resulted in Enhanced Recruitment towards Hepatocellular Carcinoma [J].
Bayo, Juan ;
Fiore, Esteban ;
Aquino, Jorge B. ;
Malvicini, Mariana ;
Rizzo, Manglio ;
Peixoto, Estanislao ;
Andriani, Oscar ;
Alaniz, Laura ;
Piccioni, Flavia ;
Bolontrade, Marcela ;
Podhajcer, Osvaldo ;
Garcia, Mariana G. ;
Mazzolini, Guillermo .
PLOS ONE, 2014, 9 (04)
[10]   Single-step isolation of extracellular vesicles by size-exclusion chromatography [J].
Boing, Anita N. ;
van der Pol, Edwin ;
Grootemaat, Anita E. ;
Coumans, Frank A. W. ;
Sturk, Auguste ;
Nieuwland, Rienk .
JOURNAL OF EXTRACELLULAR VESICLES, 2014, 3 (01)
123456