Targeting miR-27a/VE-cadherin interactions rescues cerebral cavernous malformations in mice

PLoS Biol. 2020 Jun 5;18(6):e3000734. doi: 10.1371/journal.pbio.3000734. eCollection 2020 Jun.

Abstract

Cerebral cavernous malformations (CCMs) are vascular lesions predominantly developing in the central nervous system (CNS), with no effective treatments other than surgery. Loss-of-function mutation in CCM1/krev interaction trapped 1 (KRIT1), CCM2, or CCM3/programmed cell death 10 (PDCD10) causes lesions that are characterized by abnormal vascular integrity. Vascular endothelial cadherin (VE-cadherin), a major regulator of endothelial cell (EC) junctional integrity is strongly disorganized in ECs lining the CCM lesions. We report here that microRNA-27a (miR-27a), a negative regulator of VE-cadherin, is elevated in ECs isolated from mouse brains developing early CCM lesions and in cultured ECs with CCM1 or CCM2 depletion. Furthermore, we show miR-27a acts downstream of kruppel-like factor (KLF)2 and KLF4, two known key transcription factors involved in CCM lesion development. Using CD5-2 (a target site blocker [TSB]) to prevent the miR-27a/VE-cadherin mRNA interaction, we present a potential therapy to increase VE-cadherin expression and thus rescue the abnormal vascular integrity. In CCM1- or CCM2-depleted ECs, CD5-2 reduces monolayer permeability, and in Ccm1 heterozygous mice, it restores dermal vessel barrier function. In a neonatal mouse model of CCM disease, CD5-2 normalizes vasculature and reduces vascular leakage in the lesions, inhibits the development of large lesions, and significantly reduces the size of established lesions in the hindbrain. Furthermore, CD5-2 limits the accumulation of inflammatory cells in the lesion area. Our work has established that VE-cadherin is a potential therapeutic target for normalization of the vasculature and highlights that targeting miR-27a/VE-cadherin interaction by CD5-2 is a potential novel therapy for the devastating disease, CCM.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, CD / metabolism*
  • Cadherins / metabolism*
  • Down-Regulation / genetics
  • Hemangioma, Cavernous, Central Nervous System / genetics*
  • Hemangioma, Cavernous, Central Nervous System / pathology
  • Human Umbilical Vein Endothelial Cells / metabolism
  • Humans
  • Kruppel-Like Factor 4
  • Kruppel-Like Transcription Factors / metabolism
  • Male
  • Mice, Inbred C57BL
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Rhombencephalon / blood supply
  • Rhombencephalon / pathology
  • Up-Regulation / genetics
  • rhoA GTP-Binding Protein / metabolism

Substances

  • Antigens, CD
  • Cadherins
  • KLF4 protein, human
  • Klf4 protein, mouse
  • Kruppel-Like Factor 4
  • Kruppel-Like Transcription Factors
  • MicroRNAs
  • Mirn27 microRNA, mouse
  • cadherin 5
  • rhoA GTP-Binding Protein

Grants and funding

This work was supported by the National Health and Medical Research Council (NHMRC) of Australia, #571408 (JRG), #1074664 (JL), and #161558 (XZ), https://nhmrc.gov.au/, and the National Natural Science Foundation of China, #81873752 (ZS), http://www.nsfc.gov.cn/english/site_1/index.html. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.