A Comprehensive Resource for Induced Pluripotent Stem Cells from Patients with Primary Tauopathies

Abstract

Primary tauopathies are characterized neuropathologically by inclusions containing abnormal forms of the microtubule-associated protein tau (MAPT) and clinically by diverse neuropsychiatric, cognitive, and motor impairments. Autosomal dominant mutations in the MAPT gene cause heterogeneous forms of frontotemporal lobar degeneration with tauopathy (FTLD-Tau). Common and rare variants in the MAPT gene increase the risk for sporadic FTLD-Tau, including progressive supranuclear palsy (PSP) and corticobasal degeneration (CBD). We generated a collection of fibroblasts from 140 MAPT mutation/risk variant carriers, PSP, CBD, and cognitively normal controls; 31 induced pluripotent stem cell (iPSC) lines from MAPT mutation carriers, non-carrier family members, and autopsy-confirmed PSP patients; 33 genome engineered iPSCs that were corrected or mutagenized; and forebrain neural progenitor cells (NPCs). Here, we present a resource of fibroblasts, iPSCs, and NPCs with comprehensive clinical histories that can be accessed by the scientific community for disease modeling and development of novel therapeutics for tauopathies.

Keywords: CRISPR/Cas9; MAPT; corticobasal degeneration; fibroblasts; frontotemporal dementia; induced pluripotent stem cells; neural progenitor cells; progressive supranuclear palsy; tau; tauopathy.

Graphical abstract

Figure 1

MAPT Mutations Cause Primary Tauopathy (A) Schematic of the location of MAPT mutations reported in this collection. MAPT A152T, V337M, G389R, and R406W occur in all tau isoforms expressed in the brain. MAPT P301L, P301S, and S305I/N/S occur exclusively in transcripts containing exon 10 (2N4R, 1N4R, and 0N4R). MAPT P301L/S, S305I/N/S and IVS10+16 alter splicing of tau such that more 4R-containing transcripts are expressed. (B–I) Neuropathology in human brains with primary tauopathies. (B–E) MAPT R406W carrier. (B) Atrophy of the frontal lobe with dilatation of the lateral ventricle and prominent shrinkage of the medial temporal lobe. Scale bar, 0.5 cm. (C) Neuronal loss, gliosis, and microvacuolation of superficial laminae of the superior temporal gyrus. H&E. (D) Neuronal cytoplasmic PHF1-immunoreactive inclusions are seen in the hippocampal CA1 subfield. (E) Pick body-like, PHF1-immunoreactive inclusion bodies in the dentate fascia. Scale bar in (C), (D), and (E), 50 μm. (F and G) Anterior cingulate gyrus of a MAPT V337M carrier. (F) RD4-immunoreactive cytoplasmic inclusions in spindle, also called von Economo, neurons and surrounding layer V neurons. (G) R3 (RD3) tau-immunoreactive cytoplasmic inclusions in spindle and surrounding layer V neurons, and in the neuropil. (H) Dentate gyrus of MAPT P301L case showing typical pTAU (CP13) ring-like perinuclear deposit and Pick body-like inclusions. (I) PSP associated with a MAPT A152T variant. Tufted astrocyte (left; white arrow), neurofibrillary tangle (center; open arrow), and oligodendroglial coiled bodies (right; black arrow), stained with a phospho-tau antibody (CP13). Scale bar, 25 μm.

Figure 2

Generation and Characterization of iPSC Models of Tauopathy Representative images of control (MAPT WT/WT), mutant (MAPT P301L/WT), and CRISPR/Cas9-edited, isogenic control (MAPT WT/WT-iso) iPSCs. (A) Diagram of reprogramming and CRISPR/Cas9 editing. (B and C) Immunostaining (B) and qPCR (C) for pluripotency markers. Graph represents mean ± SEM. (D) Sanger sequencing. (E) Karyotyping. (F and G) Spontaneous differentiation into cells within the three germ layers evaluated by RT-PCR (F) and immunostaining (G). MAPT WT/WT (iPSC line: F11350); MAPT P301L/WT (iPSC line: F0510); MAPT WT-iso (iPSC line: F0510.2Δ2′H1). Scale bars, 50 μm. See also Table S1.

Figure 3

Differentiation of iPSCs into Neural Progenitor Cells (A) Diagram for neural progenitor derivation protocol. (B–E) Bright-field images. (B) iPSC. (C) Neural aggregates. (D) Neural rosettes. (E) NPCs. (F) RT-PCR of neural progenitor cell markers, NESTIN, SOX2, PAX6, and the housekeeping gene, ACTIN. (G) Immunostaining for neural progenitor cell marker, PAX6. (H and I) Immunostaining of iPSC-derived neurons. (H) Tuj1. (I) MAP2. (J) Immunostaining of iPSC-derived astrocytes with GFAP. Scale bars, 50 μm.