Targeted delivery of microRNA-29b by transferrin-conjugated anionic lipopolyplex nanoparticles: a novel therapeutic strategy in acute myeloid leukemia

Clin Cancer Res. 2013 May 1;19(9):2355-67. doi: 10.1158/1078-0432.CCR-12-3191. Epub 2013 Mar 14.

Abstract

Purpose: miR-29b directly or indirectly targets genes involved in acute myeloid leukemia (AML), namely, DNMTs, CDK6, SP1, KIT, and FLT3. Higher miR-29b pretreatment expression is associated with improved response to decitabine and better outcome in AML. Thus, designing a strategy to increase miR-29b levels in AML blasts may be of therapeutic value. However, free synthetic miRs are easily degraded in bio-fluids and have limited cellular uptake. To overcome these limitations, we developed a novel transferrin-conjugated nanoparticle delivery system for synthetic miR-29b (Tf-NP-miR-29b).

Experimental design: Delivery efficiency was investigated by flow cytometry, confocal microscopy, and quantitative PCR. The expression of miR-29b targets was measured by immunoblotting. The antileukemic activity of Tf-NP-miR-29b was evaluated by measuring cell proliferation and colony formation ability and in a leukemia mouse model.

Results: Tf-NP-miR-29b treatment resulted in more than 200-fold increase of mature miR-29b compared with free miR-29b and was approximately twice as efficient as treatment with non-transferrin-conjugated NP-miR-29b. Tf-NP-miR-29b treatment significantly downregulated DNMTs, CDK6, SP1, KIT, and FLT3 and decreased AML cell growth by 30% to 50% and impaired colony formation by approximately 50%. Mice engrafted with AML cells and then treated with Tf-NP-miR-29b had significantly longer survival compared with Tf-NP-scramble (P = 0.015) or free miR-29b (P = 0.003). Furthermore, priming AML cell with Tf-NP-miR-29b before treatment with decitabine resulted in marked decrease in cell viability in vitro and showed improved antileukemic activity compared with decitabine alone (P = 0.001) in vivo.

Conclusions: Tf-NP effectively delivered functional miR-29b, resulting in target downregulation and antileukemic activity and warrants further investigation as a novel therapeutic approach in AML.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Antimetabolites, Antineoplastic / pharmacology*
  • Azacitidine / analogs & derivatives*
  • Azacitidine / pharmacology
  • Cell Line, Tumor
  • Combined Modality Therapy
  • DNA (Cytosine-5-)-Methyltransferases / genetics
  • DNA (Cytosine-5-)-Methyltransferases / metabolism
  • Decitabine
  • Down-Regulation
  • Gene Expression
  • Gene Expression Regulation, Leukemic
  • Gene Knockdown Techniques
  • Gene Transfer Techniques
  • Humans
  • Leukemia, Myeloid, Acute / therapy*
  • Linoleic Acid / chemistry
  • Male
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • MicroRNAs / genetics*
  • MicroRNAs / metabolism
  • Nanoparticles / chemistry*
  • Phosphatidylethanolamines / chemistry
  • Polyethylene Glycols / chemistry
  • RNA Interference
  • Transferrin / chemistry*
  • Xenograft Model Antitumor Assays

Substances

  • 1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine
  • Antimetabolites, Antineoplastic
  • MIRN29a microRNA, human
  • MicroRNAs
  • Phosphatidylethanolamines
  • Transferrin
  • Polyethylene Glycols
  • Decitabine
  • Linoleic Acid
  • DNA (Cytosine-5-)-Methyltransferases
  • Azacitidine