Genome-wide transcriptome study in skin biopsies reveals an association of E2F4 with cadasil and cognitive impairment

Abstract

CADASIL is a small vessel disease caused by mutations in NOTCH3 that lead to an odd number of cysteines in the EGF-like repeat domain, causing protein misfolding and aggregation. The main symptoms are migraine, psychiatric disturbances, recurrent strokes and dementia, being executive function characteristically impaired. The molecular pathways altered by this receptor aggregation need to be studied further. A genome-wide transcriptome study (four cases paired with three healthy siblings) was carried out, in addition to a qRT-PCR for validation purposes (ten new cases and eight new controls). To study the expression profile by cell type of the significant mRNAs found, we performed an in situ hybridization (ISH) (nine cases and eight controls) and a research in the Single-nuclei Brain RNA-seq expression browser (SNBREB). Pathway analysis enrichment was carried out with Gene Ontology and Reactome. Neuropsychological tests were performed in five of the qRT-PCR cases. The two most significant differentially expressed mRNAs (BANP, p-value = 7.23 × 10–4 and PDCD6IP, p-value = 8.36 × 10–4) were selected for the validation study by qRT-PCR. Additionally, we selected two more mRNAs (CAMK2G, p-value = 4.52 × 10–3 and E2F4, p-value = 4.77 × 10–3) due to their association with ischemic neuronal death. E2F4 showed differential expression in the genome-wide transcriptome study and in the qRT-PCR (p = 1.23 × 10–3), and it was upregulated in CADASIL cases. Furthermore, higher E2F4 expression was associated with worse executive function (p = 2.04 × 10–2) and attention and information processing speed (IPS) (p = 8.73 × 10–2). In situ hibridization showed E2F4 expression in endothelial and vascular smooth vessel cells. In silico studies indicated that E2F4 is also expressed in brain endothelial cells. Among the most significant pathways analyzed, there was an enrichment of vascular development, cell adhesion and vesicular machinery terms and autophagy process. E2F4 is more highly expressed in the skin biopsy of CADASIL patients compared to controls, and its expression is present in endothelial cells and VSMCs. Further studies are needed to understand whether E2F4 could be useful as a biomarker, to monitor the disease or be used as a therapeutic target.

E2F4DN Transgenic Mice: A Tool for the Evaluation of E2F4 as a Therapeutic Target in Neuropathology and Brain Aging

“…As observed in this cell line, as well as in mouse embryonic stem cells, E2F4 may activate or repress gene expression, according to its interaction partner [27,19]. Interestingly, E2F4 malfunction has been linked to cognitive impairment [94] as well as the ethiopathology of neurodegenerative diseases such as Alzheimer's disease (AD) [11,12]. This is consistent with a recent study that proposes E2F4 as a major regulator of most AD-specific gene networks [95], and with other bioinformatics-based studies suggesting that E2F4 participates in this disease [147][148][149].…”

“…E2F4 has been related to cognitive impairment [94] and the pathogenesis of AD [95] as well as of other neurological diseases [96], including Parkinson´s disease/mild cognitive impairment [97]. Since AD is largely a synaptic failure [98], occurring previous to cognitive decline or cell death [99], it can be speculated that E2F4 is important for synaptic function.…”

Role of CGRP pathway polymorphisms in migraine: a systematic review and impact on CGRP mAbs migraine therapy

“…A window is opening on the association between disabling primary headache and preclinical familial Alzheimer’s disease [ 64 ]. Moreover, both dementia and migraine are main symptoms of CADASIL (cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy, OMIM#125,310) [ 65 ]. Therefore, the genetic association of these widespread disabling diseases needs to be studied.…”

Identification of Dysregulated Mechanisms and Potential Biomarkers in Ischemic Stroke Onset

“…58 Downregulation of PPIE is associated with cognitive dysfunction in IS. 51,59 Macrophages that substantially infiltrate stroke lesions in IS patients may adopt a pro-inflammatory phenotype that enhances brain damage or a catabolic inflammatory phenotype that enhances brain repair. 60 Monocytes in the blood may mediate neuroinflammatory responses that can strongly affect IS outcomes, and these cells may serve as the basis of new therapeutic approaches.…”