Cellular Functions and Gene and Protein Expression Profiles in Endothelial Cells Derived from Moyamoya Disease-Specific iPS Cells

Abstract

Background and purposeMoyamoya disease (MMD) is a slow, progressive steno-occlusive disease, arising in the terminal portions of the cerebral internal carotid artery. However, the functions and characteristics of the endothelial cells (ECs) in MMD are unknown. We analyzed these features using induced pluripotent stem cell (iPSC)-derived ECs.MethodsiPSC lines were established from the peripheral blood of three patients with MMD carrying the variant RNF213 R4810K, and three healthy persons used as controls. After the endothelial differentiation of iPSCs, CD31+CD144+ cells were purified as ECs using a cell sorter. We analyzed their proliferation, angiogenesis, and responses to some angiogenic factors, namely VEGF, bFGF, TGF-β, and BMP4. The ECs were also analyzed using DNA microarray and proteomics to perform comprehensive gene and protein expression analysis.ResultsAngiogenesis was significantly impaired in MMD regardless of the presence of any angiogenic factor. On the contrary, endothelial proliferation was not significant between control- and MMD-derived cells. Regarding DNA microarray, pathway analysis illustrated that extracellular matrix (ECM) receptor-related genes, including integrin β3, were significantly downregulated in MMD. Proteomic analysis revealed that cytoskeleton-related proteins were downregulated and splicing regulation-related proteins were upregulated in MMD.ConclusionsDownregulation of ECM receptor-related genes may be associated with impaired angiogenic activity in ECs derived from iPSCs from patients with MMD. Upregulation of splicing regulation-related proteins implied differences in splicing patterns between control and MMD ECs.

Dysregulation of RNF213 promotes cerebral hypoperfusion

“…At present, there is no clear explanation as to why arteriogenesis is inhibited in the KO mice but not in the EC-specific RNF213 p.R4810K variant. While further study is essential to explain this difference, it is of interest to note that a reduced angiogenesis phenotype was found in vivo in both KO and EC-Tg mice, and consistently confirmed by several independent groups using different biological specimens, such as iPS-derived ECs or endothelial progenitor cells from patients with MMD 17 , 38 , 39 , as well as cultured ECs overexpressing different RNF213 variants found in Asian or Caucasian patients 18 . A reduced CBF, which is one of the characteristics of MMD patients 8 , is also manifest in both KO and EC-Tg mice.…”

The application of convolutional neural network to stem cell biology

“…iPSCs can differentiate into mature endothelial cells [57] and can be applied in disease modeling and organ formation. iPSC-derived endothelial cells (iPSC-ECs) have been used to ameliorate the cellular pathology of Moyamoya disease [58], aortic valve calcification [59], and pulmonary arterial hypertension [11]. The initial step in iPSC research is to identify iPSC-derived cells and check their quality by microscopic observation.…”

Cerebrovascular disorders associated with genetic lesions

“…The most recently identified candidate gene, CCER2 on chromosome 19, has been proposed as a potential biomarker of MMD due to brain-specific expression [177]. Additional insight into MMD will be obtained by establishing and investigating patient-derived cell lines and by analysis of coding as well as non-coding RNA, which could also provide potential therapeutic targets [178-180]. …”