Friday, October 21, 2016

Oligodendrocytes induce motor neuron death in ALS

Ferraiuolo L, Meyer K, Sherwood TW, Vick J, Likhite S, Frakes A, Miranda CJ,
Braun L, Heath PR, Pineda R, Beattie CE, Shaw PJ, Askwith CC, McTigue D, Kaspar
BK. Oligodendrocytes contribute to motor neuron death in ALS via SOD1-dependent
mechanism. Proc Natl Acad Sci U S A. 2016 Sep 29. pii: 201607496. [Epub ahead of

Oligodendrocytes have recently been implicated in the pathophysiology of amyotrophic lateral sclerosis (ALS). Here we show that, in vitro, mutant superoxide dismutase 1 (SOD1) mouse oligodendrocytes induce WT motor neuron (MN) hyperexcitability and death. Moreover, we efficiently derived human oligodendrocytes from a large number of controls and patients with sporadic and familial ALS, using two different reprogramming methods. All ALS oligodendrocyte lines induced MN death through conditioned medium (CM) and in coculture. CM-mediated MN death was associated with decreased lactate production and release, whereas toxicity in coculture was lactate-independent, demonstrating that MN survival is mediated not only by soluble factors. Remarkably, human SOD1 shRNA treatment resulted in MN rescue in both mouse and human cultures when knockdown was achieved in progenitor cells, whereas it was ineffective in differentiated oligodendrocytes. In fact, early SOD1 knockdown rescued lactate impairment and cell toxicity in all lines tested, with the exclusion of samples carrying chromosome 9 ORF 72 (C9orf72) repeat expansions. These did not respond to SOD1 knockdown nor did they show lactate release impairment. Our data indicate that SOD1 is directly or indirectly involved in ALS oligodendrocyte pathology and suggest that in this cell type, some damage might be irreversible. In addition, we demonstrate that patients with C9ORF72 represent an independent patient group that might not respond to the same treatment.

A first-of-its-kind oligodendrocyte in vitro model shows that human cells normally supportive of motor neuron function play an active role in amyotrophic lateral sclerosis pathogenesis – and this discovery may point the way toward therapeutic timing and targets.

A number of studies over the last decade have shown that cells which normally support motor neurons, such as astrocytes and microglia, contribute to motor neuron death during the progression of amyotrophic lateral sclerosis (ALS). More recently, work with mouse models has shown that oligodendrocytes, also important to normal motor neuron function, are involved in ALS progression as well…

“We were able to dig deep in trying to make a bridge between a mouse model and what is happening in humans,” says Brian Kaspar, PhD, senior author of the paper and a principal investigator in the Center for Gene Therapy at The Research Institute at Nationwide Children’s. “We have been able to begin asking questions about how exactly oligodendrocytes go wrong and how they lead to motor neuron death.”

The researchers developed a novel co-culture model that allowed observation of mouse and human ALS oligodendrocytes and their effect on motor neurons. In addition, the study partially relied on a “direct conversion” method pioneered in Dr. Kaspar’s laboratory, which allows the programming of skin cells from living ALS patients to become neural progenitor cells.

A previous study by Dr. Kaspar’s lab already showed that these skin-derived neuronal progenitor cells can efficiently be differentiated into astrocytes; the current study proves that the cells can become oligodendrocytes and presumably other cell types as well…

Mutations of the gene SOD1 have long been implicated in familial ALS, but studies from multiple laboratories including Dr. Kaspar’s provided evidence that the gene may also have a role to play in sporadic ALS. The new study found that reducing levels of the SOD1 enzyme in progenitor cells before they become oligodendrocytes reduced the toxicity towards motor neurons leading to either greater survival or complete rescue. That was true in all familial and sporadic ALS samples with the exception of samples that carried C9orf72 gene mutations.

After the progenitors fully matured into oligodendrocytes, however, SOD1 knockdown did not result in motor neuron rescue.

“There is a point of no return,” explains Dr. Kaspar, who is also a professor in the Department of Pediatrics and Department of Neuroscience at The Ohio State University College of Medicine. “When these cells have reached a certain point of differentiation or in their toxicity profile, you can’t reverse it. This tells us to treat as early as possible, which is an emerging theme in many neurodegenerative disorders.”

Direct conversion is an easier and faster method for deriving cells than the more traditional reprogramming to induced pluripotent stem cells (iPSCs). Moreover, there is increasing evidence that direct conversion allows the age-profile of a cell to be maintained, which is crucial when studying adult-onset neurodegenerative disorders, says Kathrin Meyer, PhD, a researcher in Dr. Kaspar’s lab.

“The method is fast enough that we can derive and test these cells while a patient is still alive and eligible for clinical trials,” Dr. Meyer says. “We now have even more proof that this method works, and we can use it to look at disease progression or at more specific differences between patient populations, thus developing more targeted disease treatments.”

1 comment:

  1. Meyer K, Ferraiuolo L, Miranda CJ, Likhite S, McElroy S, Renusch S, Ditsworth D, Lagier-Tourenne C, Smith RA, Ravits J, Burghes AH, Shaw PJ, Cleveland DW, Kolb SJ, Kaspar BK. Direct conversion of patient fibroblasts demonstrates non-cell autonomous toxicity of astrocytes to motor neurons in familial and sporadic ALS. Proc Natl Acad Sci U S A. 2014 Jan 14;111(2):829-32.

    Amyotrophic lateral sclerosis (ALS) causes motor neuron degeneration, paralysis, and death. Accurate disease modeling, identifying disease mechanisms, and developing therapeutics is urgently needed. We previously reported motor neuron toxicity through postmortem ALS spinal cord-derived astrocytes. However, these cells can only be harvested after death, and their expansion is limited. We now report a rapid, highly reproducible method to convert adult human fibroblasts from living ALS patients to induced neuronal progenitor cells and subsequent differentiation into astrocytes (i-astrocytes). Non-cell autonomous toxicity to motor neurons is found following coculture of i-astrocytes from familial ALS patients with mutation in superoxide dismutase or hexanucleotide expansion in C9orf72 (ORF 72 on chromosome 9) the two most frequent causes of ALS. Remarkably, i-astrocytes from sporadic ALS patients are as toxic as those with causative mutations, suggesting a common mechanism. Easy production and expansion of i-astrocytes now enables rapid disease modeling and high-throughput drug screening to alleviate astrocyte-derived toxicity.