Abstract

Preeclampsia is a placentally induced hypertensive disorder of pregnancy that is associated with substantial morbidity and mortality to mothers and fetuses. Clinical manifestations of preterm preeclampsia result from excess circulating soluble vascular endothelial growth factor receptor FLT1 (sFLT1 or sVEGFR1) of placental origin. Here we identify short interfering RNAs (siRNAs) that selectively silence the three sFLT1 mRNA isoforms primarily responsible for placental overexpression of sFLT1 without reducing levels of full-length FLT1 mRNA. Full chemical stabilization in the context of hydrophobic modifications enabled productive siRNA accumulation in the placenta (up to 7% of injected dose) and reduced circulating sFLT1 in pregnant mice (up to 50%). In a baboon preeclampsia model, a single dose of siRNAs suppressed sFLT1 overexpression and clinical signs of preeclampsia. Our results demonstrate RNAi-based extrahepatic modulation of gene expression with nonformulated siRNAs in nonhuman primates and establish a path toward a new treatment paradigm for patients with preterm preeclampsia.

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Acknowledgements

We thank D. Conte for help revising the manuscript and Khvorova Laboratory members for their support. This project was funded by the National Institutes of Health (R01 HD086111 and S10 OD020012) and the Bill and Melinda Gates Foundation (OPP1086170).

Author information

Author notes

    • Anton A Turanov
    • , Agnes Lo
    • , Matthew R Hassler
    •  & Angela Makris

    These authors contributed equally to this work.

Affiliations

  1. RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, Massachusetts, USA.

    • Anton A Turanov
    • , Matthew R Hassler
    • , Ami Ashar-Patel
    • , Julia F Alterman
    • , Andrew H Coles
    • , Reka A Haraszti
    • , Loic Roux
    • , Bruno M D C Godinho
    • , Dimas Echeverria
    • , Melissa J Moore
    •  & Anastasia Khvorova
  2. Center for Vascular Biology Research, Beth Israel Deaconess Medical Center, Boston, Massachusetts, USA.

    • Agnes Lo
    • , Zsuzsanna K Zsengeller
    •  & S Ananth Karumanchi
  3. Harvard Medical School, Boston, Massachusetts, USA.

    • Agnes Lo
    • , Zsuzsanna K Zsengeller
    •  & S Ananth Karumanchi
  4. Heart Research Institute, Sydney, New South Wales, Australia.

    • Angela Makris
    • , Suzanne Pears
    • , Renuka Shanmugalingam
    •  & Annemarie Hennessy
  5. School of Medicine, Western Sydney University, Sydney, New South Wales, Australia.

    • Angela Makris
    • , Jim Iliopoulos
    • , Renuka Shanmugalingam
    •  & Annemarie Hennessy
  6. Renal Department, Liverpool Hospital, Sydney, New South Wales, Australia.

    • Angela Makris
    •  & Renuka Shanmugalingam
  7. Women's and Babies, Royal Prince Alfred Hospital, Sydney, New South Wales, Australia.

    • Robert Ogle
  8. Department of Medicine, Cedars-Sinai Medical Center, Los Angeles, California, USA.

    • S Ananth Karumanchi
  9. Moderna Therapeutics, Cambridge, Massachusetts, USA.

    • Melissa J Moore
  10. Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts.

    • Anastasia Khvorova

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Contributions

A.A.T. managed the project, performed most of the experiments and drafted the manuscript. M.J.M., S.A.K. and A.K. conceived the study and led the project. M.R.H., D.E. and L.R. synthesized all compounds and developed and optimized protocols for gram-scale synthesis. A.H.C. and B.M.D.C.G. helped with in vivo studies. R.A.H. performed PNA assay analysis. A.A.-P. performed polyadenylation site sequence analysis and an initial siRNA screen. J.F.A. helped with the siRNA screen. A.L. performed primary cytotrophoblast isolations, ELISAs, toxicity studies and mouse pregnancy studies. Z.K.Z. performed placenta vascular immunohistochemistry. A.H., A.M., S.P., J.I., R.S. and R.O. developed the baboon preeclampsia model and performed all baboon experiments. A.A.T, A.K., M.J.M., S.A.K. and A.H. wrote the manuscript.

Competing interests

A.K. discloses ownership of stock in RXi Pharmaceuticals and Advirna. S.A.K. is a consultant to Thermofisher Scientific and owns stock in Aggamin Therapeutics. M.J.M. is employed by Moderna Therapeutics.

Corresponding authors

Correspondence to S Ananth Karumanchi or Melissa J Moore or Anastasia Khvorova.

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    Chemical modification patterns and sequences of siRNAs

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https://doi.org/10.1038/nbt.4297

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