Genetically engineered and enucleated human mesenchymal stromal cells for the targeted delivery of therapeutics to diseased tissue.
Journal
Nature biomedical engineering
ISSN: 2157-846X
Titre abrégé: Nat Biomed Eng
Pays: England
ID NLM: 101696896
Informations de publication
Date de publication:
07 2022
07 2022
Historique:
received:
03
11
2020
accepted:
07
07
2021
pubmed:
22
12
2021
medline:
20
7
2022
entrez:
21
12
2021
Statut:
ppublish
Résumé
Targeting the delivery of therapeutics specifically to diseased tissue enhances their efficacy and decreases their side effects. Here we show that mesenchymal stromal cells with their nuclei removed by density-gradient centrifugation following the genetic modification of the cells for their display of chemoattractant receptors and endothelial-cell-binding molecules are effective vehicles for the targeted delivery of therapeutics. The enucleated cells neither proliferate nor permanently engraft in the host, yet retain the organelles for energy and protein production, undergo integrin-regulated adhesion to inflamed endothelial cells, and actively home to chemokine gradients established by diseased tissues. In mouse models of acute inflammation and of pancreatitis, systemically administered enucleated cells expressing two types of chemokine receptor and an endothelial adhesion molecule enhanced the delivery of an anti-inflammatory cytokine to diseased tissue (with respect to unmodified stromal cells and to exosomes derived from bone-marrow-derived stromal cells), attenuating inflammation and ameliorating disease pathology. Enucleated cells retain most of the cells' functionality, yet acquire the cargo-carrying characteristics of cell-free delivery systems, and hence represent a versatile delivery vehicle and therapeutic system.
Identifiants
pubmed: 34931077
doi: 10.1038/s41551-021-00815-9
pii: 10.1038/s41551-021-00815-9
pmc: PMC9207157
mid: NIHMS1722979
doi:
Substances chimiques
Chemokines
0
Cytokines
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
882-897Subventions
Organisme : NIH HHS
ID : P40 OD011050
Pays : United States
Organisme : NINDS NIH HHS
ID : P30 NS047101
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA184594
Pays : United States
Organisme : NCI NIH HHS
ID : U54 CA210184
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA097022
Pays : United States
Organisme : NIH HHS
ID : T32 OD017863
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA182495
Pays : United States
Organisme : NCI NIH HHS
ID : P30 CA023100
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL082792
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM137605
Pays : United States
Informations de copyright
© 2021. The Author(s), under exclusive licence to Springer Nature Limited.
Références
Allen, T. M. & Cullis, P. R. Drug delivery systems: entering the mainstream. Science 303, 1818–1822 (2004).
pubmed: 15031496
doi: 10.1126/science.1095833
Yoo, J. W., Irvine, D. J., Discher, D. E. & Mitragotri, S. Bio-inspired, bioengineered and biomimetic drug delivery carriers. Nat. Rev. Drug Discov. 10, 521–535 (2011).
pubmed: 21720407
doi: 10.1038/nrd3499
Lyerly, H. K., Osada, T. & Hartman, Z. C. Right time and place for IL12: targeted delivery stimulates immune therapy. Clin. Cancer Res. 25, 9–11 (2019).
pubmed: 30377197
doi: 10.1158/1078-0432.CCR-18-2819
Fioranelli, M. & Roccia, M. G. Twenty-five years of studies and trials for the therapeutic application of IL-10 immunomodulating properties. From high doses administration to low dose medicine new paradigm. J. Integr. Cardiol. 1, 2–6 (2014).
Samanta, S. et al. Exosomes: new molecular targets of diseases. Acta Pharmacol. Sin. 39, 501–513 (2018).
pubmed: 29219950
doi: 10.1038/aps.2017.162
Han, X., Wang, C. & Liu, Z. Red blood cells as smart delivery systems. Bioconjugate Chem. 29, 852–860 (2018).
doi: 10.1021/acs.bioconjchem.7b00758
Fang, R. H., Kroll, A. V., Gao, W. & Zhang, L. Cell membrane coating nanotechnology. Adv. Mater. 30, e1706759 (2018).
pubmed: 29582476
pmcid: 5984176
doi: 10.1002/adma.201706759
Thanuja, M. Y., Anupama, C. & Ranganath, S. H. Bioengineered cellular and cell membrane-derived vehicles for actively targeted drug delivery: so near and yet so far. Adv. Drug Deliv. Rev. 132, 57–80 (2018).
doi: 10.1016/j.addr.2018.06.012
Labusca, L., Herea, D. D. & Mashayekhi, K. Stem cells as delivery vehicles for regenerative medicine-challenges and perspectives. World J. Stem Cells 10, 43–56 (2018).
pubmed: 29849930
pmcid: 5973910
doi: 10.4252/wjsc.v10.i5.43
Sackstein, R. The lymphocyte homing receptors: gatekeepers of the multistep paradigm. Curr. Opin. Hematol. 12, 444–450 (2005).
pubmed: 16217160
doi: 10.1097/01.moh.0000177827.78280.79
Nitzsche, F. et al. Concise review: MSC adhesion cascade–insights into homing and transendothelial migration. Stem Cells 35, 1446–1460 (2017).
pubmed: 28316123
doi: 10.1002/stem.2614
Ullah, M., Liu, D. D. & Thakor, A. S. Mesenchymal stromal cell homing: mechanisms and strategies for improvement. iScience 15, 421–438 (2019).
pubmed: 31121468
pmcid: 6529790
doi: 10.1016/j.isci.2019.05.004
Fischer, U. M. et al. Pulmonary passage is a major obstacle for intravenous stem cell delivery: the pulmonary first-pass effect. Stem Cells Dev. 18, 683–692 (2009).
pubmed: 19099374
doi: 10.1089/scd.2008.0253
Karp, J. M. & Leng Teo, G. S. Mesenchymal stem cell homing: the devil is in the details. Cell Stem Cell 4, 206–216 (2009).
pubmed: 19265660
doi: 10.1016/j.stem.2009.02.001
Galipeau, J. & Sensebe, L. Mesenchymal stromal cells: clinical challenges and therapeutic opportunities. Cell Stem Cell 22, 824–833 (2018).
pubmed: 29859173
pmcid: 6434696
doi: 10.1016/j.stem.2018.05.004
Marks, P. W., Witten, C. M. & Califf, R. M. Clarifying stem-cell therapy’s benefits and risks. N. Engl. J. Med. 376, 1007–1009 (2017).
pubmed: 27959704
doi: 10.1056/NEJMp1613723
Wigler, M. H. & Weinstein, I. B. A preparative method for obtaining enucleated mammalian cells. Biochem. Biophys. Res. Commun. 63, 669–674 (1975).
pubmed: 1169063
doi: 10.1016/S0006-291X(75)80436-0
Shay, J. W. Cell enucleation, cybrids, reconstituted cells, and nuclear hybrids. Methods Enzymol. 151, 221–237 (1987).
pubmed: 3501528
doi: 10.1016/S0076-6879(87)51020-5
Coimbra, V. C. et al. Enucleated L929 cells support invasion, differentiation, and multiplication of Trypanosoma cruzi parasites. Infect. Immun. 75, 3700–3706 (2007).
pubmed: 17502387
pmcid: 1951981
doi: 10.1128/IAI.00194-07
Graham, D. M. et al. Enucleated cells reveal differential roles of the nucleus in cell migration, polarity, and mechanotransduction. J. Cell Biol. 217, 895–914 (2018).
pubmed: 29351995
pmcid: 5839789
doi: 10.1083/jcb.201706097
Wolbank, S. et al. Telomerase immortalized human amnion- and adipose-derived mesenchymal stem cells: maintenance of differentiation and immunomodulatory characteristics. Tissue Eng. Part A 15, 1843–1854 (2009).
pubmed: 19125642
doi: 10.1089/ten.tea.2008.0205
Malawista, S. E., Van Blaricom, G. & Breitenstein, M. G. Cryopreservable neutrophil surrogates. Stored cytoplasts from human polymorphonuclear leukocytes retain chemotactic, phagocytic, and microbicidal function. J. Clin. Invest. 83, 728–732 (1989).
pubmed: 2536406
pmcid: 303736
doi: 10.1172/JCI113939
Keys, J., Windsor, A. & Lammerding, J. Assembly and use of a microfluidic device to study cell migration in confined environments. Methods Mol. Biol. 1840, 101–118 (2018).
pubmed: 30141042
doi: 10.1007/978-1-4939-8691-0_10
Lammerding, J. Mechanics of the nucleus. Compr. Physiol. 1, 783–807 (2011).
pubmed: 23737203
pmcid: 4600468
doi: 10.1002/cphy.c100038
Guilak, F., Tedrow, J. R. & Burgkart, R. Viscoelastic properties of the cell nucleus. Biochem. Biophys. Res. Commun. 269, 781–786 (2000).
pubmed: 10720492
doi: 10.1006/bbrc.2000.2360
Stewart-Hutchinson, P. J., Hale, C. M., Wirtz, D. & Hodzic, D. Structural requirements for the assembly of LINC complexes and their function in cellular mechanical stiffness. Exp. Cell. Res. 314, 1892–1905 (2008).
pubmed: 18396275
pmcid: 2562747
doi: 10.1016/j.yexcr.2008.02.022
Caille, N., Thoumine, O., Tardy, Y. & Meister, J. J. Contribution of the nucleus to the mechanical properties of endothelial cells. J. Biomech. 35, 177–187 (2002).
pubmed: 11784536
doi: 10.1016/S0021-9290(01)00201-9
Marquez-Curtis, L. A. & Janowska-Wieczorek, A. Enhancing the migration ability of mesenchymal stromal cells by targeting the SDF-1/CXCR4 axis. BioMed. Res. Int. 2013, 561098 (2013).
pubmed: 24381939
pmcid: 3870125
doi: 10.1155/2013/561098
Pandolfi, F. et al. Integrins: integrating the biology and therapy of cell–cell interactions. Clin. Ther. 39, 2420–2436 (2017).
pubmed: 29203050
doi: 10.1016/j.clinthera.2017.11.002
Chigaev, A. et al. Real time analysis of the affinity regulation of alpha 4-integrin. The physiologically activated receptor is intermediate in affinity between resting and Mn(2+) or antibody activation. J. Biol. Chem. 276, 48670–48678 (2001).
pubmed: 11641394
doi: 10.1074/jbc.M103194200
Boltze, J. et al. The dark side of the force – constraints and complications of cell therapies for stroke. Front. Neurol. 6, 155 (2015).
pubmed: 26257702
pmcid: 4507146
doi: 10.3389/fneur.2015.00155
Jung, J. W. et al. Familial occurrence of pulmonary embolism after intravenous, adipose tissue-derived stem cell therapy. Yonsei Med. J. 54, 1293–1296 (2013).
pubmed: 23918585
pmcid: 3743204
doi: 10.3349/ymj.2013.54.5.1293
Bartosh, T. J. et al. Aggregation of human mesenchymal stromal cells (MSCs) into 3D spheroids enhances their antiinflammatory properties. Proc. Natl Acad. Sci. USA 107, 13724–13729 (2010).
pubmed: 20643923
pmcid: 2922230
doi: 10.1073/pnas.1008117107
Levy, O. et al. mRNA-engineered mesenchymal stem cells for targeted delivery of interleukin-10 to sites of inflammation. Blood 122, e23–e32 (2013).
pubmed: 23980067
pmcid: 3790516
doi: 10.1182/blood-2013-04-495119
Gordon, S. et al. Antigen markers of macrophage differentiation in murine tissues. Curr. Top. Microbiol. Immunol. 181, 1–37 (1992).
pubmed: 1424778
Devine, M. J., Mierisch, C. M., Jang, E., Anderson, P. C. & Balian, G. Transplanted bone marrow cells localize to fracture callus in a mouse model. J. Orthop. Res. 20, 1232–1239 (2002).
pubmed: 12472234
doi: 10.1016/S0736-0266(02)00051-7
Ignowski, J. M. & Schaffer, D. V. Kinetic analysis and modeling of firefly luciferase as a quantitative reporter gene in live mammalian cells. Biotechnol. Bioeng. 86, 827–834 (2004).
pubmed: 15162459
doi: 10.1002/bit.20059
Alvarez, H. M. et al. Effects of PEGylation and immune complex formation on the pharmacokinetics and biodistribution of recombinant interleukin 10 in mice. Drug Metab. Dispos. 40, 360–373 (2012).
pubmed: 22083830
doi: 10.1124/dmd.111.042531
Greenberg, J. A. et al. Clinical practice guideline: management of acute pancreatitis. Can. J. Surg. 59, 128–140 (2016).
pubmed: 27007094
pmcid: 4814287
doi: 10.1503/cjs.015015
Forsmark, C. E., Vege, S. S. & Wilcox, C. M. Acute Pancreatitis. N. Engl. J. Med. 375, 1972–1981 (2016).
pubmed: 27959604
doi: 10.1056/NEJMra1505202
Niederau, C., Ferrell, L. D. & Grendell, J. H. Caerulein-induced acute necrotizing pancreatitis in mice: protective effects of proglumide, benzotript, and secretin. Gastroenterology 88, 1192–1204 (1985).
pubmed: 2984080
doi: 10.1016/S0016-5085(85)80079-2
Su, K. H., Cuthbertson, C. & Christophi, C. Review of experimental animal models of acute pancreatitis. HPB 8, 264–286 (2006).
pubmed: 18333137
doi: 10.1080/13651820500467358
Rongione, A. J. et al. Interleukin 10 reduces the severity of acute pancreatitis in rats. Gastroenterology 112, 960–967 (1997).
pubmed: 9041259
doi: 10.1053/gast.1997.v112.pm9041259
van Laethem, J. L. et al. Interleukin 10 prevents necrosis in murine experimental acute pancreatitis. Gastroenterology 108, 1917–1922 (1995).
pubmed: 7539389
doi: 10.1016/0016-5085(95)90158-2
Fedorak, R. N. et al. Recombinant human interleukin 10 in the treatment of patients with mild to moderately active Crohn’s disease. The Interleukin 10 Inflammatory Bowel Disease Cooperative Study Group. Gastroenterology 119, 1473–1482 (2000).
pubmed: 11113068
doi: 10.1053/gast.2000.20229
Nowakowski, A., Andrzejewska, A., Janowski, M., Walczak, P. & Lukomska, B. Genetic engineering of stem cells for enhanced therapy. Acta Neurobiol. Exp. 73, 1–18 (2013).
Cui, L. L. et al. The cerebral embolism evoked by intra-arterial delivery of allogeneic bone marrow mesenchymal stem cells in rats is related to cell dose and infusion velocity. Stem Cell Res. Ther. 6, 11 (2015).
pubmed: 25971703
pmcid: 4429328
doi: 10.1186/scrt544
Krueger, T. E. G., Thorek, D. L. J., Denmeade, S. R., Isaacs, J. T. & Brennen, W. N. Concise review: mesenchymal stem cell-based drug delivery: the good, the bad, the ugly, and the promise. Stem Cells Transl. Med. 7, 651–663 (2018).
pubmed: 30070053
pmcid: 6127224
doi: 10.1002/sctm.18-0024
Yin, J. Q., Zhu, J. & Ankrum, J. A. Manufacturing of primed mesenchymal stromal cells for therapy. Nat. Biomed. Eng. 3, 90–104 (2019).
pubmed: 30944433
doi: 10.1038/s41551-018-0325-8
Ungerechts, G. et al. Moving oncolytic viruses into the clinic: clinical-grade production, purification, and characterization of diverse oncolytic viruses. Mol. Ther. Methods Clin. Dev. 3, 16018 (2016).
pubmed: 27088104
pmcid: 4822647
doi: 10.1038/mtm.2016.18
Chen, H., Marino, S. & Ho, C. Y. 97. Large scale purification of AAV with continuous flow ultracentrifugation. Mol. Ther. 24, S42 (2016).
doi: 10.1016/S1525-0016(16)32906-9
Vazquez-Lombardi, R., Roome, B. & Christ, D. Molecular engineering of therapeutic cytokines. Antibodies 2, 426–451 (2013).
doi: 10.3390/antib2030426
Wigler, M. H., Neugut, A. I. & Weinstein, I. B. Enucleation of mammalian cells in suspension. Methods Cell. Biol. 14, 87–93 (1976).
pubmed: 794633
doi: 10.1016/S0091-679X(08)60471-9
Bartosh, T. J. & Ylostalo, J. H. Preparation of anti-inflammatory mesenchymal stem/precursor cells (MSCs) through sphere formation using hanging-drop culture technique. Curr. Protoc. Stem Cell Biol. 28, Unit 2B.6 (2014).
pubmed: 24510769
pmcid: 3998749
doi: 10.1002/9780470151808.sc02b06s28
Frith, J. E., Thomson, B. & Genever, P. G. Dynamic three-dimensional culture methods enhance mesenchymal stem cell properties and increase therapeutic potential. Tissue Eng. Part C. Methods 16, 735–749 (2010).
pubmed: 19811095
doi: 10.1089/ten.tec.2009.0432
Egger, D., Tripisciano, C., Weber, V., Dominici, M. & Kasper, C. Dynamic cultivation of mesenchymal stem cell aggregates. Bioengineering 5, 48 (2018).
pmcid: 6026937
doi: 10.3390/bioengineering5020048
Davidson, P. M., Sliz, J., Isermann, P., Denais, C. & Lammerding, J. Design of a microfluidic device to quantify dynamic intra-nuclear deformation during cell migration through confining environments. Integr. Biol. 7, 1534–1546 (2015).
doi: 10.1039/C5IB00200A
Hyduk, S. J. et al. Talin-1 and kindlin-3 regulate alpha4beta1 integrin-mediated adhesion stabilization, but not G protein-coupled receptor-induced affinity upregulation. J. Immunol. 187, 4360–4368 (2011).
pubmed: 21911599
doi: 10.4049/jimmunol.1003725
Semon, J. A. et al. Integrin expression and integrin-mediated adhesion in vitro of human multipotent stromal cells (MSCs) to endothelial cells from various blood vessels. Cell Tissue Res. 341, 147–158 (2010).
pubmed: 20563599
doi: 10.1007/s00441-010-0994-4
Quah, B. J., Warren, H. S. & Parish, C. R. Monitoring lymphocyte proliferation in vitro and in vivo with the intracellular fluorescent dye carboxyfluorescein diacetate succinimidyl ester. Nat. Protoc. 2, 2049–2056 (2007).
pubmed: 17853860
doi: 10.1038/nprot.2007.296
Jing, D. et al. Tissue clearing of both hard and soft tissue organs with the PEGASOS method. Cell Res. 28, 803–818 (2018).
pubmed: 29844583
pmcid: 6082844
doi: 10.1038/s41422-018-0049-z
Corradetti, B. et al. Hyaluronic acid coatings as a simple and efficient approach to improve MSC homing toward the site of inflammation. Sci. Rep. 7, 7991 (2017).
pubmed: 28801676
pmcid: 5554184
doi: 10.1038/s41598-017-08687-3