Non-viral gene delivery utilizing RALA modulates sFlt-1 secretion, important for preeclampsia
Abstract
Background: Overexpression of sFlt-1 or modulation of FKBPL, key antiangiogenic proteins, are important in the pathogenesis of preeclampsia. Methods: A newly developed nonviral gene-delivery system, RALA, capable of overexpressing sFlt-1 (e15a isoform) was delivered in vivo in transgenic haploinsufficient (Fkbpl+/−) mice. RALA was also used in vitro to deliver human Flt1 (hFlt1) in trophoblast cells. Results: Serum stable and nontoxic RALA/DNA-based nanoparticles induced an increase in sFlt-1 protein levels in the blood and total protein in the urine; the effect was more pronounced in Fkbpl+/− mice. In vitro, RALA-hFlt nanoparticles significantly reduced secretion of sFlt-1 in trophoblast cells. Conclusion: The RALA-based genetic nanodelivery system can be safely and effectively applied to emulate preeclampsia-like features or reduce sFlt-1 levels in vitro.
Lay abstract
In this study, the investigators utilized a safe and effective approach to modulate an important circulating protein in pregnancy, sFlt-1, associated with the pregnancy complication, preeclampsia. Preeclampsia is a complex and multifactorial disease and a leading cause of death in pregnancy with no current effective treatment strategies. This is likely due to a lack of reliable preclinical models that replicate human disease. The authors demonstrate the feasibility of a new preeclampsia-like model based on the dysfunction of two key vascular proteins, sFlt-1 and FKBPL (an important protein involved in the development of new blood vessels), that could be utilized in the future for testing and development of new treatments targeting these important mechanisms in preeclampsia.
Tweetable abstract
Development of a new model of #preeclampsia using a serum-stable and safe nonviral gene delivery system, #RALA, to modulate #sFlt-1 secretion in #FKBPL knockdown mice in vivo or #trophoblasts in vitro.
Graphical abstract
Papers of special note have been highlighted as: • of interest; •• of considerable interest
References
- 1. . Preeclampsia. Circ. Res. 124(7), 1094–1112 (2019).
- 2. . Elucidating the pathogenesis of pre-eclampsia using in vitro models of spiral uterine artery remodeling. Curr. Hypertens. Rep. 19(11), 93 (2017).
- 3. . The two stage model of preeclampsia: variations on the theme. Placenta 30(Suppl. A), S32–S37 (2009).
- 4. . Mechanisms of key innate immune cells in early- and late-onset preeclampsia. Front. Immunol. 11 (2020). • This review provides a comprehensive outline of distinct and overlapping mechanisms regulating different phenotypes of preeclampsia contributing to the body of knowledge and a better understanding of preeclampsia.
- 5. . Inhibition of vascular endothelial cell growth factor activity by an endogenously encoded soluble receptor. Proc. Natl Acad. Sci. 90(22), 10705–10709 (1993).
- 6. Excess placental soluble fms-like tyrosine kinase 1 (sFlt1) may contribute to endothelial dysfunction, hypertension, and proteinuria in preeclampsia. J. Clin. Invest. 111(5), 649–658 (2003). •• This is one of the first studies that demonstrated the importance of sFlt-1 in inducing preeclampsia symptoms and features in vivo.
- 7. Anti-angiogenic factors and pre-eclampsia in type 1 diabetic women. Diabetologia 52(1), 160–168 (2009).
- 8. Circulating angiogenic factors and the risk of preeclampsia. N. Engl. J. Med. 350(7), 672–683 (2004).
- 9. Predictive value of the sFlt-1:PlGF ratio in women with suspected preeclampsia. N. Engl. J. Med. 374(1), 13–22 (2016).
- 10. Removal of soluble Fms-like tyrosine kinase-1 by dextran sulfate apheresis in preeclampsia. J. Am. Soc. Nephrol. 27(3), 903–913 (2016).
- 11. A Novel human-specific soluble vascular endothelial growth factor receptor 1. Circ. Res. 102(12), 1566–1574 (2008).
- 12. A recently evolved novel trophoblast-enriched secreted form of fms-like tyrosine kinase-1 variant is up-regulated in hypoxia and preeclampsia. J. Clin. Endocrinol. Metab. 94(7), 2524–2530 (2009). •• Here, a specific variant of sFlt-1 (e15a) was identified as the main isoform enriched in placentae with potential application for preeclampsia development.
- 13. . Role of biomarkers in early detection of preeclampsia. J. Clin. Diagnostic Res. 8(4), BE01 (2014).
- 14. FKBPL and peptide derivatives: novel biological agents that inhibit angiogenesis by a CD44-dependent mechanism. Clin. Cancer Res. (2011).
- 15. FKBPL is a critical antiangiogenic regulator of developmental and pathological angiogenesis. Arterioscler. Thromb. Vasc. Biol. 35(4), 845–854 (2015). •• This is the first report that demonstrates the importance of FKBPL in developmental and physiological angiogenesis and the development of Fkbpl haploinsufficient mouse model.
- 16. . The therapeutic and diagnostic potential of FKBPL; A novel anticancer protein. Drug Discov. Today 11-12, 544–8 (2012).
- 17. . Trophoblast lineage specification, differentiation and their regulation by oxygen tension. J. Endocrinol. 236(1), R43–R56 (2018).
- 18. . Placental trophoblast cell differentiation: physiological regulation and pathological relevance to preeclampsia. Mol. Aspects Med. 34(5), 981–1023 (2013).
- 19. Targeting treatment-resistant breast cancer stem cells with FKBPL and its peptide derivative, AD-01, via the CD44 pathway. Clin. Cancer Res. 19(14), 3881–3893 (2013).
- 20. FKBPL and its peptide derivatives inhibit endocrine therapy resistant cancer stem cells and breast cancer metastasis by downregulating DLL4 and Notch4. BMC Cancer. 19(1), 351 (2019).
- 21. Role of A Novel angiogenesis FKBPL-CD44 pathway in preeclampsia risk stratification and mesenchymal stem cell treatment. J. Clin. Endocrinol. Metab. 106(1), 26–41 (2021). •• This is the first report to demonstrate the predictive and diagnostic biomarker potential of FKBPL and its target protein, CD44, in preeclampsia. FKBPL's role as a target of potential treatment for preeclampsia based on mesenchymal stem cells was also described.
- 22. FKBPL is associated with metabolic parameters and is a novel determinant of cardiovascular disease. Sci. Rep. 10(1), 21655 (2020).
- 23. . Postpartum interventions to reduce long-term cardiovascular disease risk in women after hypertensive disorders of pregnancy: a systematic review. Front. Cardiovasc. Med. 6, 160 (2019).
- 24. Considerations to model heart disease in women with preeclampsia and cardiovascular disease. Cells. 10(4), 899 (2021). •• This comprehensive review critically evaluates current preclinical models of preeclampsia and discusses the need for more reliable and personalized models of this disease. It particularly emphasizes the need for preclinical models that recapitulate future cardiovascular disease and endothelial dysfunction as a result of preeclampsia in pregnancy.
- 25. . Animal models of preeclampsia: translational failings and why. Am. J. Physiol. Integr. Comp. Physiol. 314(4), R499–R508 (2018).
- 26. . Recent advances in nanomaterials for gene delivery – a review. Nanomaterials 7(5), 94 (2017).
- 27. Reprogramming human T cell function and specificity with non-viral genome targeting. Nature 559(7714), 405–409 (2018).
- 28. Development and characterization of self-assembling nanoparticles using a bio-inspired amphipathic peptide for gene delivery. J. Control. Release 189, 141–149 (2014). •• This is the first report of a novel nonviral gene-delivery system, RALA, that fully characterizes its potential for safe and effective gene delivery in vitro and in vivo.
- 29. . Delivery of RALA/siFKBPL nanoparticles via electrospun bilayer nanofibres: an innovative angiogenic therapy for wound repair. J. Control. Release. 316, 53–65 (2019).
- 30. . Protein kinase C regulates FLT1 abundance and stimulates its cleavage in vascular endothelial cells with the release of a soluble PlGF/VEGF antagonist. Exp. Cell Res. 319(17), 2578–2587 (2013).
- 31. RALA-mediated delivery of FKBPL nucleic acid therapeutics. Nanomedicine (Lond). 10(19), 2989–3001 (2015). • RALA is efficient at delivering FKBPL as a therapeutic in vivo to treat cancer. This study validated RALA's potential for safe and effective delivery of nucleic acid therapeutics.
- 32. The effect of over-expression of sFlt-1 on blood pressure and the occurrence of other manifestations of preeclampsia in unrestrained conscious pregnant mice. Am. J. Obstet. Gynecol. 196(4), 396.e1–396.e7 (2007).
- 33. Full-length human placental sFlt-1-e15a isoform induces distinct maternal phenotypes of preeclampsia in mice. PLoS ONE 10(4), e0119547 (2015).
- 34. Systemic RALA/iNOS nanoparticles: a potent gene therapy for metastatic breast cancer coupled as a biomarker of treatment. Mol. Ther. Nucleic Acids 6, 249–258 (2017).
- 35. Gene therapy with RALA/iNOS composite nanoparticles significantly enhances survival in a model of metastatic prostate cancer. Cancer Nanotechnol. 9(1), 5 (2018).
- 36. Proteinuria in preeclampsia: not essential to diagnosis but related to disease severity and fetal outcomes. Pregnancy Hypertens. 8, 60–64 (2017).
- 37. . Animal models of preeclampsia. Semin. Nephrol. 24(6), 596–606 (2004).
- 38. . Animal models of pre-eclampsia. Am. J. Reprod. Immunol. 65(6), 533–541 (2011).
- 39. . Vascular endothelial growth factor and angiogenesis. Pharmacol. Rev. 56(4), 549–580 (2004).
- 40. . In vivo rat model of preeclampsia. Methods Mol. Med. 122, 393–399 (2006).
- 41. Soluble endoglin contributes to the pathogenesis of preeclampsia. Nat. Med. 12(6), 642–649 (2006).
- 42. Pravastatin induces placental growth factor (PGF) and ameliorates preeclampsia in a mouse model. Proc. Natl Acad. Sci. U. S. A. 108(4), 1451–1455 (2011).
- 43. Reduction of circulating soluble Flt-1 alleviates preeclampsia-like symptoms in a mouse model. J. Cell. Mol. Med. 14(6B), 1857–1867 (2010).
- 44. Current model systems for the study of preeclampsia. Exp. Biol. Med. (Maywood). 243(6), 576–585 (2018).
- 45. . The promise and potential hazards of adenovirus gene therapy. Gut 48(5), 733–736 (2001).
- 46. DNA vaccination for cervical cancer: strategic optimisation of RALA mediated gene delivery from a biodegradable microneedle system. Eur. J. Pharm. Biopharm. 127, 288–297 (2018).
- 47. RNAi modulation of placental sFLT1 for the treatment of preeclampsia. Nat. Biotechnol. 36(12), 1164–1173 (2018).