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Lipid nanoparticle-mediated siRNA delivery for safe targeting of human CML in vivo

  • Nidhi Jyotsana
  • Amit Sharma
  • Anuhar Chaturvedi
  • Ramachandramouli Budida
  • Michaela Scherr
  • Florian Kuchenbauer
  • Robert Lindner
  • Fatih Noyan
  • Kurt-Wolfram Sühs
  • Martin Stangel
  • Denis Grote-Koska
  • Korbinian Brand
  • Hans-Peter Vornlocher
  • Matthias Eder
  • Felicitas Thol
  • Arnold Ganser
  • R. Keith Humphries
  • Euan Ramsay
  • Pieter Cullis
  • Michael HeuserEmail author
Original Article

Abstract

Efficient and safe delivery of siRNA in vivo is the biggest roadblock to clinical translation of RNA interference (RNAi)-based therapeutics. To date, lipid nanoparticles (LNPs) have shown efficient delivery of siRNA to the liver; however, delivery to other organs, especially hematopoietic tissues still remains a challenge. We developed DLin-MC3-DMA lipid-based LNP-siRNA formulations for systemic delivery against a driver oncogene to target human chronic myeloid leukemia (CML) cells in vivo. A microfluidic mixing technology was used to obtain reproducible ionizable cationic LNPs loaded with siRNA molecules targeting the BCR-ABL fusion oncogene found in CML. We show a highly efficient and non-toxic delivery of siRNA in vitro and in vivo with nearly 100% uptake of LNP-siRNA formulations in bone marrow of a leukemic model. By targeting the BCR-ABL fusion oncogene, we show a reduction of leukemic burden in our myeloid leukemia mouse model and demonstrate reduced disease burden in mice treated with LNP-BCR-ABL siRNA as compared with LNP-CTRL siRNA. Our study provides proof-of-principle that fusion oncogene specific RNAi therapeutics can be exploited against leukemic cells and promise novel treatment options for leukemia patients.

Keywords

Lipid nanoparticle BCR-ABL RNAi Chronic myeloid leukemia 

Notes

Acknowledgements

We thank Silke Glowotz, Martin Wichmann, Anitha Thomas and Colin Walsh for technical support. We express sincere thanks to the patients for their participation in the study. We thank Prof. Dr. Alf Lamprecht and Dr. Manusmriti Singh for providing us nanoparticles during the initial experiments. We thank the staff of the Central Animal Facility and Matthias Ballmaier from the Cell Sorting Core Facility (supported in part by the Braukmann-Wittenberg-Herz-Stiftung and the Deutsche Forschungsgemeinschaft) of Hannover Medical School.

Authors’ contributions

N.J. and M.H. designed the research; N.J., A.S., A.C., M.S., R.B., M.S., F.K., R.L., F.N., K.W. S., M.S., D.G.K., K.B., H.P.V., M.E., A.G., F.T., R.K.H., E.R., P.C., and M.H. performed the research; N.J., A.S., and M.H. analyzed the data. N.J. and M.H. wrote the manuscript. All authors read and agreed to the final version of the manuscript.

Funding information

This study was supported by the Rudolf-Bartling Stiftung, an ERC grant under the European Union’s Horizon 2020 research and innovation program (No. 638035), grant 111,267 from Deutsche Krebshilfe, DFG grant HE 5240/5-2 and HE 5240/6-2; grants from Dieter-Schlag Stiftung and a Terry Fox Foundation Program Project Award to RKH.

Compliance with ethical standards

Conflict of interest

Euan Ramsay is an employee of Precision Nanosystems. Pieter Cullis is founder of Precision Nanosystems. The other authors have no conflicts of interest.

Supplementary material

277_2019_3713_MOESM1_ESM.docx (3.9 mb)
ESM 1 (DOCX 3987 kb)

References

  1. 1.
    De Kouchkovsky I, Abdul-Hay M (2016) Acute myeloid leukemia: a comprehensive review and 2016 update. Blood Cancer J 6(7):e441Google Scholar
  2. 2.
    Coelho T et al (2013) Safety and efficacy of RNAi therapy for transthyretin amyloidosis. N Engl J Med 369(9):819–829Google Scholar
  3. 3.
    Fitzgerald K, White S, Borodovsky A, Bettencourt BR, Strahs A, Clausen V, Wijngaard P, Horton JD, Taubel J, Brooks A, Fernando C, Kauffman RS, Kallend D, Vaishnaw A, Simon A (2017) A highly durable RNAi therapeutic inhibitor of PCSK9. N Engl J Med 376(1):41–51Google Scholar
  4. 4.
    Solomon SD, Adams D, Kristen A, Grogan M, González-Duarte A, Maurer MS, Merlini G, Damy T, Slama MS, Brannagan TH III, Dispenzieri A, Berk JL, Shah AM, Garg P, Vaishnaw A, Karsten V, Chen J, Gollob J, Vest J, Suhr O (2019) Effects of patisiran, an RNA interference therapeutic, on cardiac parameters in patients with hereditary transthyretin-mediated amyloidosis. Circulation 139(4):431–443Google Scholar
  5. 5.
    Lorenzer C, Dirin M, Winkler AM, Baumann V, Winkler J (2015) Going beyond the liver: progress and challenges of targeted delivery of siRNA therapeutics. J Control Release 203:1–15Google Scholar
  6. 6.
    Rungta RL, Choi HB, Lin PJC, Ko RWY, Ashby D, Nair J, Manoharan M, Cullis PR, MacVicar BA (2013) Lipid nanoparticle delivery of siRNA to silence neuronal gene expression in the brain. Mol Ther Nucleic Acids 2:e136Google Scholar
  7. 7.
    Jyotsana N, Sharma A, Chaturvedi A, Scherr M, Kuchenbauer F, Sajti L, Barchanski A, Lindner R, Noyan F, Sühs KW, Grote-Koska D, Brand K, Vornlocher HP, Stanulla M, Bornhauser B, Bourquin JP, Eder M, Thol F, Ganser A, Humphries RK, Ramsay E, Cullis P, Heuser M (2018) RNA interference efficiently targets human leukemia driven by a fusion oncogene in vivo. Leukemia 32(1):224–226Google Scholar
  8. 8.
    Belliveau NM et al (2012) Microfluidic synthesis of highly potent limit-size lipid nanoparticles for in vivo delivery of siRNA. Mol Ther Nucleic Acids 1:e37Google Scholar
  9. 9.
    Semple SC, Klimuk SK, Harasym TO, Dos Santos N, Ansell SM, Wong KF, Maurer N, Stark H, Cullis PR, Hope MJ, Scherrer P (2001) Efficient encapsulation of antisense oligonucleotides in lipid vesicles using ionizable aminolipids: formation of novel small multilamellar vesicle structures. Biochim Biophys Acta 1510(1–2):152–166Google Scholar
  10. 10.
    Hafez IM, Cullis PR (2001) Roles of lipid polymorphism in intracellular delivery. Adv Drug Deliv Rev 47(2–3):139–148Google Scholar
  11. 11.
    Tam YY, Chen S, Cullis PR (2013) Advances in lipid nanoparticles for siRNA delivery. Pharmaceutics 5(3):498–507Google Scholar
  12. 12.
    Weinstein S, Toker IA, Emmanuel R, Ramishetti S, Hazan-Halevy I, Rosenblum D, Goldsmith M, Abraham A, Benjamini O, Bairey O, Raanani P, Nagler A, Lieberman J, Peer D (2016) Harnessing RNAi-based nanomedicines for therapeutic gene silencing in B-cell malignancies. Proc Natl Acad Sci U S A 113(1):E16–E22Google Scholar
  13. 13.
    Rowley JD (1973) Letter: a new consistent chromosomal abnormality in chronic myelogenous leukaemia identified by quinacrine fluorescence and Giemsa staining. Nature 243(5405):290–293Google Scholar
  14. 14.
    Druker BJ (2002) Perspectives on the development of a molecularly targeted agent. Cancer Cell 1(1):31–36Google Scholar
  15. 15.
    Talpaz M, Shah NP, Kantarjian H, Donato N, Nicoll J, Paquette R, Cortes J, O’Brien S, Nicaise C, Bleickardt E, Blackwood-Chirchir MA, Iyer V, Chen TT, Huang F, Decillis AP, Sawyers CL (2006) Dasatinib in imatinib-resistant Philadelphia chromosome-positive leukemias. N Engl J Med 354(24):2531–2541Google Scholar
  16. 16.
    Kurzrock R, Kantarjian HM, Druker BJ, Talpaz M (2003) Philadelphia chromosome-positive leukemias: from basic mechanisms to molecular therapeutics. Ann Intern Med 138(10):819–830Google Scholar
  17. 17.
    Laurent E et al (2001) The BCR gene and Philadelphia chromosome-positive leukemogenesis. Cancer Res 61(6):2343–2355Google Scholar
  18. 18.
    Gorre ME, Mohammed M, Ellwood K, Hsu N, Paquette R, Rao PN, Sawyers CL (2001) Clinical resistance to STI-571 cancer therapy caused by BCR-ABL gene mutation or amplification. Science 293(5531):876–880Google Scholar
  19. 19.
    Yamamoto M, Kurosu T, Kakihana K, Mizuchi D, Miura O (2004) The two major imatinib resistance mutations E255K and T315I enhance the activity of BCR/ABL fusion kinase. Biochem Biophys Res Commun 319(4):1272–1275Google Scholar
  20. 20.
    Weisberg E, Griffin JD (2000) Mechanism of resistance to the ABL tyrosine kinase inhibitor STI571 in BCR/ABL-transformed hematopoietic cell lines. Blood 95(11):3498–3505Google Scholar
  21. 21.
    Branford S et al (2002) High frequency of point mutations clustered within the adenosine triphosphate-binding region of BCR/ABL in patients with chronic myeloid leukemia or Ph-positive acute lymphoblastic leukemia who develop imatinib (STI571) resistance. Blood 99(9):3472–3475Google Scholar
  22. 22.
    Scherr M, Battmer K, Winkler T, Heidenreich O, Ganser A, Eder M (2003) Specific inhibition of bcr-abl gene expression by small interfering RNA. Blood 101(4):1566–1569Google Scholar
  23. 23.
    Thomas M, Gessner A, Vornlocher HP, Hadwiger P, Greil J, Heidenreich O (2005) Targeting MLL-AF4 with short interfering RNAs inhibits clonogenicity and engraftment of t(4;11)-positive human leukemic cells. Blood 106(10):3559–3566Google Scholar
  24. 24.
    Koldehoff M, Steckel NK, Beelen DW, Elmaagacli AH (2007) Therapeutic application of small interfering RNA directed against bcr-abl transcripts to a patient with imatinib-resistant chronic myeloid leukaemia. Clin Exp Med 7(2):47–55Google Scholar
  25. 25.
    Koldehoff M (2015) Targeting bcr-abl transcripts with siRNAs in an imatinib-resistant chronic myeloid leukemia patient: challenges and future directions. Methods Mol Biol 1218:277–292Google Scholar
  26. 26.
    Valencia-Serna J, Aliabadi HM, Manfrin A, Mohseni M, Jiang X, Uludag H (2018) siRNA/lipopolymer nanoparticles to arrest growth of chronic myeloid leukemia cells in vitro and in vivo. Eur J Pharm Biopharm 130:66–70Google Scholar
  27. 27.
    Urbinati G, Ali HM, Rousseau Q, Chapuis H, Desmaële D, Couvreur P, Massaad-Massade L (2015) Antineoplastic effects of siRNA against TMPRSS2-ERG junction oncogene in prostate cancer. PLoS One 10(5):e0125277Google Scholar
  28. 28.
    Takenaka S, Naka N, Araki N, Hashimoto N, Ueda T, Yoshioka K, Yoshikawa H, Itoh K (2010) Downregulation of SS18-SSX1 expression in synovial sarcoma by small interfering RNA enhances the focal adhesion pathway and inhibits anchorage-independent growth in vitro and tumor growth in vivo. Int J Oncol 36(4):823–831Google Scholar
  29. 29.
    Scherr M, Chaturvedi A, Battmer K, Dallmann I, Schultheis B, Ganser A, Eder M (2006) Enhanced sensitivity to inhibition of SHP2, STAT5, and Gab2 expression in chronic myeloid leukemia (CML). Blood 107(8):3279–3287Google Scholar
  30. 30.
    Heuser M, Beutel G, Krauter J, Dohner K, von Neuhoff N, Schlegelberger B, Ganser A (2006) High meningioma 1 (MN1) expression as a predictor for poor outcome in acute myeloid leukemia with normal cytogenetics. Blood 108(12):3898–3905Google Scholar
  31. 31.
    Fang YP, Wu PC, Huang YB, Tzeng CC, Chen YL, Hung YH, Tsai MJ, Tsai YH (2012) Modification of polyethylene glycol onto solid lipid nanoparticles encapsulating a novel chemotherapeutic agent (PK-L4) to enhance solubility for injection delivery. Int J Nanomedicine 7:4995–5005Google Scholar
  32. 32.
    de Fougerolles A, Vornlocher HP, Maraganore J, Lieberman J (2007) Interfering with disease: a progress report on siRNA-based therapeutics. Nat Rev Drug Discov 6(6):443–453Google Scholar
  33. 33.
    Testa U, Pelosi E, Frankel A (2014) CD 123 is a membrane biomarker and a therapeutic target in hematologic malignancies. Biomark Res 2(1):4Google Scholar
  34. 34.
    Zeijlemaker W, Kelder A, Oussoren-Brockhoff YJM, Scholten WJ, Snel AN, Veldhuizen D, Cloos J, Ossenkoppele GJ, Schuurhuis GJ (2016) A simple one-tube assay for immunophenotypical quantification of leukemic stem cells in acute myeloid leukemia. Leukemia 30(2):439–446Google Scholar
  35. 35.
    Dutta S, Saxena R (2014) The expression pattern of CD33 antigen can differentiate leukemic from Normal progenitor cells in acute myeloid leukemia. Indian J Hematol Blood Transfus 30(2):130–134Google Scholar
  36. 36.
    Landesman-Milo D, Goldsmith M, Leviatan Ben-Arye S, Witenberg B, Brown E, Leibovitch S, Azriel S, Tabak S, Morad V, Peer D (2013) Hyaluronan grafted lipid-based nanoparticles as RNAi carriers for cancer cells. Cancer Lett 334(2):221–227Google Scholar
  37. 37.
    Yan XD et al (2005) The role of apolipoprotein E in the elimination of liposomes from blood by hepatocytes in the mouse. Biochem Biophys Res Commun 328(1):57–62Google Scholar
  38. 38.
    Akinc A et al (2010) Targeted delivery of RNAi therapeutics with endogenous and exogenous ligand-based mechanisms. Mol Ther 18(7):1357–1364Google Scholar
  39. 39.
    Zhou PX et al (2010) Uptake of synthetic low density lipoprotein by leukemic stem cells - a potential stem cell targeted drug delivery strategy. J Control Release 148(3):380–387Google Scholar
  40. 40.
    Wiemels JL, Cazzaniga G, Daniotti M, Eden OB, Addison GM, Masera G, Saha V, Biondi A, Greaves MF (1999) Prenatal origin of acute lymphoblastic leukaemia in children. Lancet 354(9189):1499–1503Google Scholar
  41. 41.
    Ma X et al (2015) Rise and fall of subclones from diagnosis to relapse in pediatric B-acute lymphoblastic leukaemia. Nat Commun 6:6604Google Scholar
  42. 42.
    Mullighan CG, Phillips LA, Su X, Ma J, Miller CB, Shurtleff SA, Downing JR (2008) Genomic analysis of the clonal origins of relapsed acute lymphoblastic leukemia. Science 322(5906):1377–1380Google Scholar
  43. 43.
    Hunger SP, Mullighan CG (2015) Acute lymphoblastic leukemia in children. N Engl J Med 373(16):1541–1552Google Scholar
  44. 44.
    Bernt KM, Hunger SP (2014) Current concepts in pediatric Philadelphia chromosome-positive acute lymphoblastic leukemia. Front Oncol 4:54Google Scholar
  45. 45.
    Ley TJ et al (2013) Genomic and epigenomic landscapes of adult de novo acute myeloid leukemia. N Engl J Med 368(22):2059–2074Google Scholar
  46. 46.
    O’Hare T, Corbin AS, Druker BJ (2006) Targeted CML therapy: controlling drug resistance, seeking cure. Curr Opin Genet Dev 16(1):92–99Google Scholar
  47. 47.
    Wright JR, Ung YC, Julian JA, Pritchard KI, Whelan TJ, Smith C, Szechtman B, Roa W, Mulroy L, Rudinskas L, Gagnon B, Okawara GS, Levine MN (2007) Randomized, double-blind, placebo-controlled trial of erythropoietin in non-small-cell lung cancer with disease-related anemia. J Clin Oncol 25(9):1027–1032Google Scholar
  48. 48.
    Hong DS, Kurzrock R, Oh Y, Wheler J, Naing A, Brail L, Callies S, Andre V, Kadam SK, Nasir A, Holzer TR, Meric-Bernstam F, Fishman M, Simon G (2011) A phase 1 dose escalation, pharmacokinetic, and pharmacodynamic evaluation of eIF-4E antisense oligonucleotide LY2275796 in patients with advanced cancer. Clin Cancer Res 17(20):6582–6591Google Scholar
  49. 49.
    Jyotsana N, Heuser M (2018) Exploiting differential RNA splicing patterns: a potential new group of therapeutic targets in cancer. Expert Opin Ther Targets 22(2):107–121Google Scholar
  50. 50.
    Tefferi A, Lasho TL, Begna KH, Patnaik MM, Zblewski DL, Finke CM, Laborde RR, Wassie E, Schimek L, Hanson CA, Gangat N, Wang X, Pardanani A (2015) A pilot study of the telomerase inhibitor Imetelstat for myelofibrosis. N Engl J Med 373(10):908–919Google Scholar
  51. 51.
    Geisbert TW, Lee ACH, Robbins M, Geisbert JB, Honko AN, Sood V, Johnson JC, de Jong S, Tavakoli I, Judge A, Hensley LE, MacLachlan I (2010) Postexposure protection of non-human primates against a lethal Ebola virus challenge with RNA interference: a proof-of-concept study. Lancet 375(9729):1896–1905Google Scholar
  52. 52.
    Huang X, Schwind S, Santhanam R, Eisfeld AK, Chiang CL, Lankenau M, et al (2016) Targeting the RAS/MAPK pathway with miR-181a in acute myeloid leukemia. Oncotarget 7(37):59273-86.  https://doi.org/10.18632/oncotarget.11150
  53. 53.
    Dorrance AM, Neviani P, Ferenchak GJ, Huang X, Nicolet D, Maharry KS, Ozer HG, Hoellarbauer P, Khalife J, Hill EB, Yadav M, Bolon BN, Lee RJ, Lee LJ, Croce CM, Garzon R, Caligiuri MA, Bloomfield CD, Marcucci G (2015) Targeting leukemia stem cells in vivo with antagomiR-126 nanoparticles in acute myeloid leukemia. Leukemia 29(11):2143–2153Google Scholar
  54. 54.
    Huang X, Schwind S, Yu B, Santhanam R, Wang H, Hoellerbauer P, Mims A, Klisovic R, Walker AR, Chan KK, Blum W, Perrotti D, Byrd JC, Bloomfield CD, Caligiuri MA, Lee RJ, Garzon R, Muthusamy N, Lee LJ, Marcucci G (2013) Targeted delivery of microRNA-29b by transferrin-conjugated anionic lipopolyplex nanoparticles: a novel therapeutic strategy in acute myeloid leukemia. Clin Cancer Res 19(9):2355–2367Google Scholar
  55. 55.
    Jiang X, Hu C, Arnovitz S, Bugno J, Yu M, Zuo Z, Chen P, Huang H, Ulrich B, Gurbuxani S, Weng H, Strong J, Wang Y, Li Y, Salat J, Li S, Elkahloun AG, Yang Y, Neilly MB, Larson RA, le Beau MM, Herold T, Bohlander SK, Liu PP, Zhang J, Li Z, He C, Jin J, Hong S, Chen J (2016) miR-22 has a potent anti-tumour role with therapeutic potential in acute myeloid leukaemia. Nat Commun 7:11452Google Scholar
  56. 56.
    Jiang X, Bugno J, Hu C, Yang Y, Herold T, Qi J, Chen P, Gurbuxani S, Arnovitz S, Strong J, Ferchen K, Ulrich B, Weng H, Wang Y, Huang H, Li S, Neilly MB, Larson RA, le Beau MM, Bohlander SK, Jin J, Li Z, Bradner JE, Hong S, Chen J (2016) Eradication of acute myeloid leukemia with FLT3 ligand-targeted miR-150 nanoparticles. Cancer Res 76(15):4470–4480Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Nidhi Jyotsana
    • 1
  • Amit Sharma
    • 1
  • Anuhar Chaturvedi
    • 1
  • Ramachandramouli Budida
    • 2
  • Michaela Scherr
    • 1
  • Florian Kuchenbauer
    • 3
  • Robert Lindner
    • 4
  • Fatih Noyan
    • 5
  • Kurt-Wolfram Sühs
    • 6
  • Martin Stangel
    • 6
  • Denis Grote-Koska
    • 7
  • Korbinian Brand
    • 7
  • Hans-Peter Vornlocher
    • 8
  • Matthias Eder
    • 1
  • Felicitas Thol
    • 1
  • Arnold Ganser
    • 1
  • R. Keith Humphries
    • 9
    • 10
  • Euan Ramsay
    • 11
  • Pieter Cullis
    • 12
  • Michael Heuser
    • 1
    Email author
  1. 1.Department of Hematology, Hemostasis, Oncology and Stem Cell TransplantationHannover Medical SchoolHannoverGermany
  2. 2.Department of Immunology and RheumatologyHannover Medical SchoolHannoverGermany
  3. 3.Department of Internal Medicine IIIUniversity Hospital of UlmUlmGermany
  4. 4.Department of Cell Biology, Center of AnatomyHannover Medical SchoolHannoverGermany
  5. 5.Department of Gastroenterology, Hepatology & EndocrinologyHannover Medical SchoolHannoverGermany
  6. 6.Clinic for NeurologyHannover Medical SchoolHannoverGermany
  7. 7.Department of Clinical ChemistryHannover Medical SchoolHannoverGermany
  8. 8.Axolabs GmBHKulmbachGermany
  9. 9.Terry Fox LaboratoryBritish Columbia Cancer AgencyVancouverCanada
  10. 10.Department of MedicineUniversity of British ColumbiaVancouverCanada
  11. 11.Precision NanoSystems IncVancouverCanada
  12. 12.Department of Biochemistry and Molecular BiologyUniversity of British ColumbiaVancouverCanada

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