Targeted siRNA nanocarrier: a platform technology for cancer treatment.

Carcinoma, Non-Small-Cell Lung / genetics Cell Line, Tumor Gene Expression Regulation, Neoplastic Humans Lung Neoplasms / genetics Male Oncogene Proteins, Fusion / genetics Protamines / genetics Proto-Oncogene Protein c-fli-1 / genetics RNA, Small Interfering / genetics RNA-Binding Protein EWS / genetics Technology Xenograft Model Antitumor Assays

Journal

Oncogene

ISSN: 1476-5594

Titre abrégé: Oncogene

Pays: England

ID NLM: 8711562

Informations de publication

Date de publication:
04 2022

Historique:

received: 21 06 2021

accepted: 09 02 2022

revised: 28 01 2022

pubmed: 28 2 2022

medline: 13 4 2022

entrez: 27 2 2022

Statut: ppublish

Résumé

The small arginine-rich protein protamine condenses complete genomic DNA into the sperm head. Here, we applied its high RNA binding capacity for spontaneous electrostatic assembly of therapeutic nanoparticles decorated with tumour-cell-specific antibodies for efficiently targeting siRNA. Fluorescence microscopy and DLS measurements of these nanocarriers revealed the formation of a vesicular architecture that requires presence of antibody-protamine, defined excess of free SMCC-protamine, and anionic siRNA to form. Only these complex nanoparticles were efficient in the treatment of non-small-cell lung cancer (NSCLC) xenograft models, when the oncogene KRAS was targeted via EGFR-mediated delivery. To show general applicability, we used the modular platform for IGF1R-positive Ewing sarcomas. Anti-IGR1R-antibodies were integrated into an antibody-protamine nanoparticle with an siRNA specifically against the oncogenic translocation product EWS/FLI1. Using these nanoparticles, EWS/FLI1 knockdown blocked in vitro and in vivo growth of Ewing sarcoma cells. We conclude that these antibody-protamine-siRNA nanocarriers provide a novel platform technology to specifically target different cell types and yet undruggable targets in cancer therapy by RNAi.

Identifiants

DOI: 10.1038/s41388-022-02241-w PMID: 35220407 PMC: PMC8993695

pubmed: 35220407

doi: 10.1038/s41388-022-02241-w

pii: 10.1038/s41388-022-02241-w

pmc: PMC8993695

doi:

Substances chimiques

Oncogene Proteins, Fusion 0

Protamines 0

Proto-Oncogene Protein c-fli-1 0

RNA, Small Interfering 0

RNA-Binding Protein EWS 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

Pagination

2210-2224

Informations de copyright

Références

Carr JA, Silverman N. The heparin-protamine interaction. A review. J Cardiovasc Surg. 1999;40:659–66.

Mochizuki T, Olson PJ, Szlam F, Ramsay JG, Levy JH. Protamine reversal of heparin affects platelet aggregation and activated clotting time after cardiopulmonary bypass. Anesth Analg. 1998;87:781–5.

doi: 10.1213/00000539-199810000-00008

Rossmann P, Matousovic K, Horácek V. Protamine-heparin aggregates. Their fine structure, histochemistry, and renal deposition. Virchows Arch B Cell Pathol Incl Mol Pathol. 1982;40:81–98.

doi: 10.1007/BF02932853

Smull CE, Ludwig EH. Enhancement of the plaque-forming capacity of poliovirus ribonucleic acid with basic proteins. J Bacteriol. 1962;84:1035–40.

doi: 10.1128/jb.84.5.1035-1040.1962

Pardi N, Hogan MJ, Porter FW, Weissman D. mRNA vaccines—a new era in vaccinology. Nat Rev Drug Discov. 2018;17:261–79.

doi: 10.1038/nrd.2017.243

Bäumer N, Appel N, Terheyden L, Buchholz F, Rossig C, Müller-Tidow C, et al. Antibody-coupled siRNA as an efficient method for in vivo mRNA knockdown. Nat Protoc. 2016;11:22–36.

doi: 10.1038/nprot.2015.137

Bäumer N, Rehkamper J, Appel N, Terheyden L, Hartmann W, Wardelmann E, et al. Downregulation of PIK3CA via antibody-esiRNA-complexes suppresses human xenograft tumor growth. PLoS ONE. 2018;13:e0200163.

doi: 10.1371/journal.pone.0200163

Bäumer S, Bäumer N, Appel N, Terheyden L, Fremerey J, Schelhaas S, et al. Antibody-mediated delivery of Anti-KRAS-siRNA in vivo overcomes therapy resistance in colon cancer. Clin Cancer Res. 2015;21:1383–94.

doi: 10.1158/1078-0432.CCR-13-2017

Song E, Zhu P, Lee SK, Chowdhury D, Kussman S, Dykxhoorn DM, et al. Antibody mediated in vivo delivery of small interfering RNAs via cell-surface receptors. Nat Biotechnol. 2005;2005:709–17.

doi: 10.1038/nbt1101

Hed J, Hallden G, Johansson S, Larsson P. The use of fluorescence quenching in flow cytofluorometry to measure the attachment and ingestion phases in phagocytosis in peripheral blood without prior cell separation. J Immunological Methods. 1987;101:119–25.

doi: 10.1016/0022-1759(87)90224-9

Einstein A. Über die von der molekularkinetischen Theorie der Wärme geforderte Bewegung von in ruhenden Flüssigkeiten suspendierten Teilchen. Ann Phys. 1905;322:549–60.

doi: 10.1002/andp.19053220806

Doody JF, Wang Y, Patel SN, Joynes C, Lee SP, Gerlak J, et al. Inhibitory activity of cetuximab on epidermal growth factor receptor mutations in non small cell lung cancers. Mol Cancer Ther. 2007;6:2642–51.

doi: 10.1158/1535-7163.MCT-06-0506

Ladenstein R, Pötschger U, Le Deley MC, Whelan J, Paulussen M, Oberlin O, et al. Primary disseminated multifocal Ewing sarcoma: results of the Euro-EWING 99 trial. J Clin Oncol. 2010;28:3284–91.

doi: 10.1200/JCO.2009.22.9864

Delattre O, Zucman J, Plougastel B, Desmaze C, Melot T, Peter M, et al. Gene fusion with an ETS DNA-binding domain caused by chromosome translocation in human tumours. Nature. 1992;359:162–5.

doi: 10.1038/359162a0

Naing A, LoRusso P, Fu S, Hong DS, Anderson P, Benjamin RS, et al. Insulin growth factor-receptor (IGF-1R) antibody cixutumumab combined with the mTOR inhibitor temsirolimus in patients with refractory Ewing’s sarcoma family tumors. Clin Cancer Res. 2012;18:2625–31.

doi: 10.1158/1078-0432.CCR-12-0061

Dolgin E. IGF-1R drugs travel from cancer cradle to Graves’. Nat Biotechnol. 2020;38:385–8.

doi: 10.1038/s41587-020-0481-8

Dohjima T, Sook Lee N, Li H, Ohno T, Rossi JJ. Small interfering RNAs expressed from a Pol III promoter suppress the EWS/Fli-1 transcript in an Ewing sarcoma cell line. Mol Ther. 2003;7:811–6. https://www.cell.com/molecular-therapy-family/molecular-therapy/fulltext/S1525-0016(03)00101-1 .

doi: 10.1016/S1525-0016(03)00101-1

Balhorn R, Brewer L, Corzett M. DNA condensation by protamine and arginine-rich peptides: analysis of toroid stability using single DNA molecules. Mol Reprod Dev. 2000;56:230–4.

doi: 10.1002/(SICI)1098-2795(200006)56:2+<230::AID-MRD3>3.0.CO;2-V

Delgado D, Del Pozo-Rodríguez A, Solinís MÁ, Rodríguez-Gascón A. Understanding the mechanism of protamine in solid lipid nanoparticle-based lipofection: the importance of the entry pathway. Eur J Pharm Biopharm. 2011;79:495–502.

doi: 10.1016/j.ejpb.2011.06.005

Vasimalai N, John SA. Aggregation and de-aggregation of gold nanoparticles induced by polyionic drugs: spectrofluorimetric determination of picogram amounts of protamine and heparin drugs in the presence of 1000-fold concentration of major interferences. J Mater Chem B. 2013;1:5620–7.

doi: 10.1039/c3tb20991a

Yoo H, Mok H. Evaluation of multimeric siRNA conjugates for efficient protamine-based delivery into breast cancer cells. Arch Pharm Res. 2015;38:129–36.

doi: 10.1007/s12272-014-0359-8

Rotow J, Bivona TG. Understanding and targeting resistance mechanisms in NSCLC. Nat Rev Cancer. 2017;17:637–58.

doi: 10.1038/nrc.2017.84

Wang M, Herbst RS, Boshoff C. Toward personalized treatment approaches for non-small-cell lung cancer. Nat Med. 2021;27:1345–56.

doi: 10.1038/s41591-021-01450-2