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Alzheimers Res Ther. 2019 Aug 9;11(1):71. doi: 10.1186/s13195-019-0524-x.

A single-nuclei RNA sequencing study of Mendelian and sporadic AD in the human brain.

Del-Aguila JL1,2,3, Li Z1,2,3, Dube U1,2,3, Mihindukulasuriya KA1,3, Budde JP1,2,3, Fernandez MV1,2,3, Ibanez L1,2,3, Bradley J1,2,3, Wang F1,2,3, Bergmann K1,2, Davenport R1,2, Morris JC2,4,5, Holtzman DM2,4,5, Perrin RJ2,4,6, Benitez BA1,3, Dougherty J7, Cruchaga C8,9,10,11,12, Harari O13,14,15,16,17.

Author information

1
Department of Psychiatry, Washington University School of Medicine, Campus Box 8134, 425 S. Euclid Ave, BJC Institute of Health, Office: 9607, St. Louis, MO, 63110, USA.
2
Hope Center for Neurological Disorders, Washington University School of Medicine, St. Louis, MO, USA.
3
NeuroGenomics and Informatics, Department of Psychiatry, Washington University, St. Louis, MO, USA.
4
Knight Alzheimer's Disease Research Center, Washington University School of Medicine, St. Louis, MO, USA.
5
Department of Neurology, Washington University School of Medicine, St. Louis, MO, USA.
6
Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO, USA.
7
Department of Genetics, Washington University School of Medicine, St. Louis, MO, USA.
8
Department of Psychiatry, Washington University School of Medicine, Campus Box 8134, 425 S. Euclid Ave, BJC Institute of Health, Office: 9607, St. Louis, MO, 63110, USA. cruchagac@wustl.edu.
9
Hope Center for Neurological Disorders, Washington University School of Medicine, St. Louis, MO, USA. cruchagac@wustl.edu.
10
Knight Alzheimer's Disease Research Center, Washington University School of Medicine, St. Louis, MO, USA. cruchagac@wustl.edu.
11
NeuroGenomics and Informatics, Department of Psychiatry, Washington University, St. Louis, MO, USA. cruchagac@wustl.edu.
12
Department of Genetics, Washington University School of Medicine, St. Louis, MO, USA. cruchagac@wustl.edu.
13
Department of Psychiatry, Washington University School of Medicine, Campus Box 8134, 425 S. Euclid Ave, BJC Institute of Health, Office: 9607, St. Louis, MO, 63110, USA. harario@wustl.edu.
14
Hope Center for Neurological Disorders, Washington University School of Medicine, St. Louis, MO, USA. harario@wustl.edu.
15
Knight Alzheimer's Disease Research Center, Washington University School of Medicine, St. Louis, MO, USA. harario@wustl.edu.
16
NeuroGenomics and Informatics, Department of Psychiatry, Washington University, St. Louis, MO, USA. harario@wustl.edu.
17
Department of Genetics, Washington University School of Medicine, St. Louis, MO, USA. harario@wustl.edu.

Abstract

BACKGROUND:

Alzheimer's disease (AD) is the most common form of dementia. This neurodegenerative disorder is associated with neuronal death and gliosis heavily impacting the cerebral cortex. AD has a substantial but heterogeneous genetic component, presenting both Mendelian and complex genetic architectures. Using bulk RNA-seq from the parietal lobes and deconvolution methods, we previously reported that brains exhibiting different AD genetic architecture exhibit different cellular proportions. Here, we sought to directly investigate AD brain changes in cell proportion and gene expression using single-cell resolution.

METHODS:

We generated unsorted single-nuclei RNA sequencing data from brain tissue. We leveraged the tissue donated from a carrier of a Mendelian genetic mutation, PSEN1 p.A79V, and two family members who suffer from sporadic AD, but do not carry any autosomal mutations. We evaluated alternative alignment approaches to maximize the titer of reads, genes, and cells with high quality. In addition, we employed distinct clustering strategies to determine the best approach to identify cell clusters that reveal neuronal and glial cell types and avoid artifacts such as sample and batch effects. We propose an approach to cluster cells that reduces biases and enable further analyses.

RESULTS:

We identified distinct types of neurons, both excitatory and inhibitory, and glial cells, including astrocytes, oligodendrocytes, and microglia, among others. In particular, we identified a reduced proportion of excitatory neurons in the Mendelian mutation carrier, but a similar distribution of inhibitory neurons. Furthermore, we investigated whether single-nuclei RNA-seq from the human brains recapitulate the expression profile of disease-associated microglia (DAM) discovered in mouse models. We also determined that when analyzing human single-nuclei data, it is critical to control for biases introduced by donor-specific expression profiles.

CONCLUSION:

We propose a collection of best practices to generate a highly detailed molecular cell atlas of highly informative frozen tissue stored in brain banks. Importantly, we have developed a new web application to make this unique single-nuclei molecular atlas publicly available.

KEYWORDS:

Alzheimer’s disease; PSEN1; Single-nuclei RNA-seq; Web-based brain molecular atlas

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