Aging-relevant human basal forebrain cholinergic neurons as a cell model for Alzheimer’s disease

Abstract

Background Alzheimer’s disease (AD) is an adult-onset mental disorder with aging as a major risk factor. Early and progressive degeneration of basal forebrain cholinergic neurons (BFCNs) contributes substantially to cognitive impairments of AD. An aging-relevant cell model of BFCNs will critically help understand AD and identify potential therapeutics. Recent studies demonstrate that induced neurons directly reprogrammed from adult human skin fibroblasts retain aging-associated features. However, human induced BFCNs (hiBFCNs) have yet to be achieved. Methods We examined a reprogramming procedure for the generation of aging-relevant hiBFCNs through virus-mediated expression of fate-determining transcription factors. Skin fibroblasts were obtained from healthy young persons, healthy adults and sporadic AD patients. Properties of the induced neurons were examined by immunocytochemistry, qRT-PCR, western blotting, and electrophysiology. Results We established a protocol for efficient generation of hiBFCNs from adult human skin fibroblasts. They show electrophysiological properties of mature neurons and express BFCN-specific markers, such as CHAT, p75NTR, ISL1, and VACHT. As a proof-of-concept, our preliminary results further reveal that hiBFCNs from sporadic AD patients exhibit time-dependent TAU hyperphosphorylation in the soma and dysfunctional nucleocytoplasmic transport activities. Conclusions Aging-relevant BFCNs can be directly reprogrammed from human skin fibroblasts of healthy adults and sporadic AD patients. They show promises as an aging-relevant cell model for understanding AD pathology and may be employed for therapeutics identification for AD.

Chemical Replacement of Noggin with Dorsomorphin Homolog 1 for Cost-Effective Direct Neuronal Conversion

“…Our finding will be helpful to facilitate future cohort-based studies using iN technology, which is particularly useful for better understanding aspects of human neuronal cell aging (Huh et al, 2016 ; Mertens et al, 2015 ) and human age-related diseases such as ALS (Jovıˇcić et al, 2015 ), Huntington's disease (Pircs et al, 2021 ; Victor et al, 2018 ), Parkinson's disease (Drouin-Ouellet et al, 2021 ), or Alzheimer's disease (Ma et al, 2020 ; Mertens et al, 2021 ).…”

“…As a consequence, they closely resemble embryonic, but lack signatures of adultness and age, which is a limitation when modeling age-dependent diseases (Böhnke et al, 2018 ; Miller et al, 2013 ; Vera and Studer, 2015 ). As opposed to neurons derived from iPSCs, directly converted induced neurons (iNs) from human donor fibroblasts (FBs) retain many transcriptomic, molecular, metabolic, and epigenetic aging signatures (Böhnke et al, 2018 ; Chambers et al, 2009 ; Huh et al, 2016 ; Kim et al, 2018 ; Mertens et al, 2015 ; Pang et al, 2011 ), and they thus represent an attractive model system to study age-related neurological diseases in a patient- and cohort-specific manner (Drouin-Ouellet et al, 2021 ; Ma et al, 2020 ; Mertens et al, 2021 ; Victor et al, 2014 ).…”

Macrophage Infiltration Reduces Neurodegeneration and Improves Stroke Recovery after Delayed Recanalization in Rats

“…Immunofluorescence staining was performed as previously described [ 35 ]. Rats were perfused with ice-cold PBS followed by 4% paraformaldehyde.…”

A simple, flexible and high-efficiency western blot analysis for age-related human induced neurons

“…In contrast, induced pluripotent stem cells (iPSCs)-derived neurons have been reset to a fetal status absence of agingand/or disease-associated gene expression profiles [4][5][6][7]. Therefore, hiNs appear to be more proper disease models for screening therapeutics, as well as for understanding the age-related Alzheimer's disease (AD), Parkinson's disease (PD), and other neurodegenerative diseases [7][8][9][10][11][12]. The age-related and disease-specific gene expression patterns of hiNs have been mainly revealed through bioinformatical and immunocytochemical approaches [4][5][6].…”