Abstract
Amyotrophic lateral sclerosis is a progressive disease characterized by the loss of upper and lower motor neurons, leading to paralysis of voluntary muscles. About 10% of all ALS cases are familial (fALS), among which 15–20% are linked to Cu/Zn superoxide dismutase (SOD1) mutations, usually inherited in an autosomal dominant manner. To date only one FDA approved drug is available which increases survival moderately. Our understanding of ALS disease mechanisms is largely derived from rodent model studies, however due to the differences between rodents and humans, it is necessary to have humanized models for studies of disease pathogenesis as well as drug development. Therefore, we generated a comprehensive library of a total 22 of fALS patient-specific induced pluripotent stem cell (iPSC) lines. These cells were thoroughly characterized before being deposited into the library. The library of cells includes a variety of C9orf72 mutations, sod1 mutations, FUS, ANG and FIG4 mutations. Certain mutations are represented with more than one line, which allows for studies of variable genetic backgrounds. In addition, these iPSCs can be successfully differentiated to astroglia, a cell type known to play a critical role in ALS disease progression. This library represents a comprehensive resource that can be used for ALS disease modeling and the development of novel therapeutics.
Highlights
Amyotrophic lateral sclerosis (ALS), known as Lou Gehrig’s disease, is a fatal disease characterized by the loss of upper and lower motor neurons, leading to paralysis of voluntary muscles [1]
All induced pluripotent stem cell (iPSC) have been generated by using Sox2, Oct4, Klf4 and c-Myc factors delivered by retroviruses with a programming efficiency of ~0.1%
Putative iPSCs underwent a validation profile that included: 1) morphological analysis, compact colonies with clear boundary and are comprised of cells with big nucleus and large nucleoli and scant cytoplasm (Fig. 1A); 2) pluripotent gene expression by quantitative PCR, including CDH1, Dnmt3b, FoxD3, GDF3, Lefty1/2, LIN28, Nanog, Nodal, Sall4, TDGF1, TDGF1&3, TERT, Zfp42 and ZNF206, and their expression were comparable with human embryonic stem cells (hESCs) (H9 and/or H7) (Fig. 1E and S1 Fig.); 3) pluripotent marker expression by immunocytochemistry, such as Tra1–81 and SSEA4 (Fig. 1A), Tra1–60 and SSEA3, 3) silencing of transgenes and upregulation of endogenous Sox2, Oct4, Klf4 and Myc (Fig. 1B and S1 Fig.); 4) in vitro differentiation to three germ layers via embryoid body (EB) formation (Fig. 1C and S2 Fig.)
Summary
Amyotrophic lateral sclerosis (ALS), known as Lou Gehrig’s disease, is a fatal disease characterized by the loss of upper and lower motor neurons, leading to paralysis of voluntary muscles [1]. The mechanisms involved in ALS pathogenesis are largely unknown [2]. About 10% of all cases are inherited, among which about 15–20% are linked to Cu/Zn superoxide dismutase (SOD1) mutations [3] and 40% to C9orf mutations [4,5]. Other genes, such as TDP-43, PLOS ONE | DOI:10.1371/journal.pone.0118266. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript
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