Research Findings

2024.11.25

CRISPR/CasRx suppresses KRAS-induced brain arteriovenous malformation developed in postnatal brain endothelial cells in mice

▶ JCI Insight. 2024 Nov 22;9(22):e179729. doi: 10.1172/jci.insight.179729.

Saito S1,2, Nakamura Y2, Miyashita S2, Sato T2, Hoshina K2, Okada M1, Hasegawa H1, Oishi M1, Fujii Y1, Körbelin J3, Kubota Y4, Tainaka K2, Natsumeda M1, Ueno M2.

  1. Department of Neurosurgery and.
  2. Department of System Pathology for Neurological Disorders, Brain Research Institute, Niigata University.
  3. Department of Oncology, Hematology and Bone Marrow Transplantation, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
  4. Department of Anatomy, Keio University School of Medicine.

Abstract

Brain arteriovenous malformations (bAVMs) are anomalies forming vascular tangles connecting the arteries and veins, which cause hemorrhagic stroke in young adults. Current surgical approaches are highly invasive, and alternative therapeutic methods are warranted. Recent genetic studies identified KRAS mutations in endothelial cells of bAVMs; however, the underlying process leading to malformation in the postnatal stage remains unknown. Here we established a mouse model of bAVM developing during the early postnatal stage. Among 4 methods tested, mutant KRAS specifically introduced in brain endothelial cells by brain endothelial cell-directed adeno-associated virus (AAV) and endothelial cell-specific Cdh5-CreERT2 mice successfully induced bAVMs in the postnatal period. Mutant KRAS led to the development of multiple vascular tangles and hemorrhage in the brain with increased MAPK/ERK signaling and growth in endothelial cells. Three-dimensional analyses in cleared tissue revealed dilated vascular networks connecting arteries and veins, similar to human bAVMs. Single-cell RNA-Seq revealed dysregulated gene expressions in endothelial cells and multiple cell types involved in the pathological process. Finally, we employed CRISPR/CasRx to knock down mutant KRAS expression, which efficiently suppressed bAVM development. The present model reveals pathological processes that lead to postnatal bAVMs and demonstrates the efficacy of therapeutic strategies with CRISPR/CasRx.

Pathological process and therapeutic strategy in a mouse model of brain arteriovenous malformation (bAVM).

(A, B) Abnormal vasculature formed in the brain, labeled with tdTomato (A) and blue latex (B).
(C) 3D structural analyses of the vascular tangles in the cleared brain.
(D) Single-cell RNA-seq showing involvement of multiple cell types.
(E) Suppression of bAVM by CRISPR/CasRx.
Modified from Saito et al., JCI Insight 2024.

*Reprinted under the terms of the Creative Commons Attribution 4.0 International License.

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