Characteristics and Transcriptomic Analysis of Cholinergic Neurons Derived from Induced Pluripotent Stem Cells with APP Mutation in Alzheimer’s Disease
Restricted accessResearch articleFirst published online September 10, 2024
Characteristics and Transcriptomic Analysis of Cholinergic Neurons Derived from Induced Pluripotent Stem Cells with APP Mutation in Alzheimer’s Disease
The cholinergic hypothesis is one of the main theories that describe the pathogenesis of Alzheimer’s disease (AD). Cholinergic neurons degenerate early and are severely damaged in AD. Despite extensive research, the causes of cholinergic neuron damage and the underlying molecular changes remain unclear.
Objective:
This study aimed to explore the characteristics and transcriptomic changes in cholinergic neurons derived from human induced pluripotent stem cells (iPSCs) with APP mutation.
Methods:
Peripheral blood mononuclear cells from patients with AD and healthy individuals were reprogrammed into iPSCs. The iPSCs were differentiated into cholinergic neurons. Cholinergic neurons were stained, neurotoxically tested, and electrophysiologically and transcriptomically analyzed.
Results:
The iPSCs-derived cholinergic neurons from a patient with AD carrying a mutation in APP displayed enhanced susceptibility to Aβ1-42-induced neurotoxicity, characterized by severe neurotoxic effects, such as cell body coagulation and neurite fragmentation. Cholinergic neurons exhibited electrophysiological impairments and neuronal death after 21 days of culture in the AD group. Transcriptome analysis disclosed 883 differentially expressed genes (DEGs, 420 upregulated and 463 downregulated) participating in several signaling pathways implicated in AD pathogenesis. To assess the reliability of RNA sequencing, the expression of 16 target DEGs was validated using qPCR. Finally, the expression of the 8 core genes in different cell types of brain was analyzed by the AlzData database.
Conclusions:
In this study, iPSCs-derived cholinergic neurons from AD patients with APP mutations exhibit characteristics reminiscent of neurodegenerative disease. Transcriptome analysis revealed the corresponding DEGs and pathways, providing potential biomarkers and therapeutic targets for advancing AD research.
GoateA, Chartier-HarlinMC, MullanM, et al. Segregation of a missense mutation in the amyloid precursor protein gene with familial Alzheimer’s disease. Nature1991; 349: 704–706.
3.
MajdiA, Sadigh-EteghadS, Rahigh AghsanS, et al. Amyloid-β, tau, and the cholinergic system in Alzheimer’s disease: seeking direction in a tangle of clues. Rev Neurosci2020; 31: 391–413.
4.
HampelH, MesulamMM, CuelloAC, et al. The cholinergic system in the pathophysiology and treatment of Alzheimer’s disease. Brain2018; 141: 1917–1933.
5.
BlakeMG, KrawczykMC, BarattiCM, et al. Neuropharmacology of memory consolidation and reconsolidation: Insights on central cholinergic mechanisms. J Physiol Paris2014; 108: 286–291.
6.
CraigLA, HongNS and McDonaldRJ. Revisiting the cholinergic hypothesis in the development of Alzheimer’s disease. Neurosci Biobehav Rev2011; 35: 1397–1409.
7.
Ramos-RodriguezJJ, Pacheco-HerreroM, ThyssenD, et al. Rapid β-amyloid deposition and cognitive impairment after cholinergic denervation in APP/PS1 mice. J Neuropathol Exp Neurol2013; 72: 272–285.
8.
MufsonEJ, CountsSE, PerezSE, et al. Cholinergic system during the progression of Alzheimer’s disease: therapeutic implications. Expert Rev Neurother2008; 8: 1703–1718.
9.
GrotheM, HeinsenH and TeipelSJ. Atrophy of the cholinergic Basal forebrain over the adult age range and in early stages of Alzheimer’s disease. Biol Psychiatry2012; 71: 805–813.
10.
GeulaC, DunlopSR, AyalaI, et al. Basal forebrain cholinergic system in the dementias: Vulnerability, resilience, and resistance. J Neurochem2021; 158: 1394–1411.
11.
ShiY, InoueH, WuJC, et al. Induced pluripotent stem cell technology: a decade of progress. Nat Rev Drug Discov2017; 16: 115–130.
12.
PenneyJ, RalveniusWT and TsaiLH. Modeling Alzheimer’s disease with iPSC-derived brain cells. Mol Psychiatry2020; 25: 148–167.
13.
DaganH, Flashner-AbramsonE, VasudevanS, et al. Exploring Alzheimer’s disease molecular variability via calculation of personalized transcriptional signatures. Biomolecules2020; 10: 503.
14.
WeiC, LuX, ZouQ, et al. Generation and characterization of a human induced pluripotent stem cell line (XWHNi001-A) derived from an Alzheimer’s disease patient with mutation in the APP gene. Stem Cell Res2022; 60: 102690.
15.
DarmanisS, SloanSA, ZhangY, et al. A survey of human brain transcriptome diversity at the single cell level. Proc Natl Acad Sci U S A2015; 112: 7285–7290.
16.
Ferreira-VieiraTH, GuimaraesIM, SilvaFR, et al. Alzheimer’s disease: targeting the cholinergic system. Curr Neuropharmacol2016; 14: 101–115.
17.
WhitehousePJ, PriceDL, ClarkAW, et al. Alzheimer disease: evidence for selective loss of cholinergic neurons in the nucleus basalis. Ann Neurol1981; 10: 122–126.
18.
WhitehousePJ, PriceDL, StrubleRG, et al. Alzheimer’s disease and senile dementia: loss of neurons in the basal forebrain. Science1982; 215: 1237–1239.
19.
AdkinsDE, KhachaneAN, McClayJL, et al. SNP-based analysis of neuroactive ligand-receptor interaction pathways implicates PGE2 as a novel mediator of antipsychotic treatment response: data from the CATIE study. Schizophr Res2012; 135: 200–201.
20.
OuGY, LinWW and ZhaoWJ. Construction of long noncoding RNA-associated ceRNA networks reveals potential biomarkers in Alzheimer’s disease. J Alzheimers Dis2021; 82: 169–183.
21.
SunH, YangJ, LiX, et al. Identification of feature genes and pathways for Alzheimer’s disease via WGCNA and LASSO regression. Front Comput Neurosci2022; 16: 1001546.
22.
GopalakrishnanL, ChatterjeeO, RajC, et al. An assembly of galanin-galanin receptor signaling network. J Cell Commun Signal2021; 15: 269–275.
23.
ChaudharyA, SinghV, VaradwajPK, et al. Screening natural inhibitors against upregulated G-protein coupled receptors as potential therapeutics of Alzheimer’s disease. J Biomol Struct Dyn2022; 40: 673–684.
24.
MagistriM, VelmeshevD, MakhmutovaM, et al. Transcriptomics profiling of Alzheimer’s disease reveal neurovascular defects, altered amyloid-β homeostasis, and deregulated expression of long noncoding RNAs. J Alzheimers Dis2015; 48: 647–665.
25.
PrévotT and SibilleE. Altered GABA-mediated information processing and cognitive dysfunctions in depression and other brain disorders. Mol Psychiatry2021; 26: 151–167.
26.
Burgos-RamosE, Hervás-AguilarA, Aguado-LleraD, et al. Somatostatin and Alzheimer’s disease. Mol Cell Endocrinol2008; 286: 104–111.
27.
AlmeidaVN. Somatostatin and the pathophysiology of Alzheimer’s disease. Ageing Res Rev2024; 96: 102270.
28.
ParkK, LeeJ, JangHJ, et al. Optogenetic activation of parvalbumin and somatostatin interneurons selectively restores theta-nested gamma oscillations and oscillation-induced spike timing-dependent long-term potentiation impaired by amyloid β oligomers. BMC Biol2020; 18: 7.
29.
Gonzalez-HalphenD, GhelliA, IommariniL, et al. Mitochondrial complex I and cell death: a semi-automatic shotgun model. Cell Death Dis2011; 2: e222.
30.
LiangWS, ReimanEM, VallaJ, et al. Alzheimer’s disease is associated with reduced expression of energy metabolism genes in posterior cingulate neurons. Proc Natl Acad Sci U S A2008; 105: 4441–4446.
31.
SwerdlowRH. Mitochondria and cell bioenergetics: increasingly recognized components and a possible etiologic cause of Alzheimer’s disease. Antioxid Redox Signal2012; 16: 1434–1455.
32.
GuoC, SunL, ChenX, et al. Oxidative stress, mitochondrial damage and neurodegenerative diseases. Neural Regen Res2013; 8: 2003–2014.
33.
WangW, ZhaoF, MaX, et al. Mitochondria dysfunction in the pathogenesis of Alzheimer’s disease: recent advances. Mol Neurodegener2020; 15: 30.
34.
ReissAB, GulkarovS, JacobB, et al. Mitochondria in Alzheimer’s disease pathogenesis. Life (Basel)2024; 14: 196.
35.
YangK, YanY, YuA, et al. Mitophagy in neurodegenerative disease pathogenesis. Neural Regen Res2024; 19: 998–1005.
36.
BrowneSE, LinL, MattssonA, et al. Selective antibody-induced cholinergic cell and synapse loss produce sustained hippocampal and cortical hypometabolism with correlated cognitive deficits. Exp Neurol2001; 170: 36–47.
37.
KawamataJ and ShimohamaS. Association of novel and established polymorphisms in neuronal nicotinic acetylcholine receptors with sporadic Alzheimer’s disease. J Alzheimers Dis2002; 4: 71–76.
38.
GreenwoodM, GillardBT, FarrukhR, et al. Transcription factor Creb3l1 maintains proteostasis in neuroendocrine cells. Mol Metab2022; 63: 101542.
39.
BrooksC, MurphyJ, BelcastroM, et al. Farnesylation of the transducin G protein gamma subunit is a prerequisite for its ciliary targeting in rod photoreceptors. Front Mol Neurosci2018; 11: 16.
40.
Harris-WhiteME, ChuT, BalverdeZ, et al. Effects of transforming growth factor-beta (isoforms 1–3) on amyloid-beta deposition, inflammation, and cell targeting in organotypic hippocampal slice cultures. J Neurosci1998; 18: 10366–10374.
41.
ChongJR, ChaiYL, LeeJH, et al. Increased transforming growth factor β2 in the neocortex of Alzheimer’s disease and dementia with Lewy bodies is correlated with disease severity and soluble Aβ42 load. J Alzheimers Dis2017; 56: 157–166.
42.
HashimotoY, NawaM, ChibaT, et al. Transforming growth factor beta2 autocrinally mediates neuronal cell death induced by amyloid-beta. J Neurosci Res2006; 83: 1039–1047.
