New insights into Rett syndrome

A pair of papers from the lab of Fred Gage has provided new insights into the molecular and cellular processes affected in Rett syndrome. This syndrome is associated with arrested development and autistic features. It affects mainly girls, who typically show normal development until around age two, followed by a sudden and dramatic deterioration of function, regression of language skills and the emergence of autistic symptoms. It is caused mainly by mutations in the gene encoding MeCP2, which resides on the X chromosome. Complete removal of the function of this gene is effectively lethal, explaining why Rett syndrome is not observed in boys – males who inherit that mutation are not viable. Females, who have a back-up copy of the X chromosome survive but subsequently show the symptoms of the disease.

The function of the MeCP2 protein seems very far removed from the kinds of symptoms observed when it is deleted. The job of MeCP2 is to bind to DNA that carries a specific chemical tag – a methyl group – which marks DNA for repression. When MeCP2 binds, it recruits a host of other proteins which shut down that section of DNA and prevent any genes within it from being expressed. How a defect in a process that is so fundamental could result in such specific symptoms has been a mystery.

A major barrier in understanding these processes has been the inability to assay the effects of the mutation in this gene in neurons of people who carry it. After all, unlike some other cell types, one cannot easily simply extract neurons from patients. (They tend to be using them). New stem cell technologies developed over the last few years offer a way around this problem. It is possible to extract fibroblasts from patients with a simple skin biopsy. By transfecting these cells with genes that are normally expressed in embryonic stem cells it is possible to “de-differentiate” them – to turn them back into a stem cell. (The difference between a skin cell and a stem cell lies in the genes that are being expressed – transfecting the cells with the master regulatory genes that determine embryonic stem cell identity forces the expression profile back to that state). These “induced pluripotent stem cells” (iPS cells) can then be encouraged to differentiate into any of the cell-types of the body, including neurons. In this way, a virtual biopsy of a patient’s neurons can be obtained.

Gage and colleagues did exactly that, generating neurons in a dish from patients with Rett syndrome. I make that technique sound simple, but of course it isn’t, and these experiments represent a technical tour de force. They were then able to characterise various parameters of these neurons to assay more directly the molecular and cellular effects of MeCP2 mutation. These experiments revealed a not unexpected defect in the formation of synapses between Rett mutation neurons. Neurons from Rett mutation-carriers developed normally and showed normal electrophysiological properties but made fewer synapses with each other and showed a concomitant decrease in network activity. I say not unexpected because it had previously been shown that mouse neurons carrying a MeCP2 mutation show similar effects. This fits with highly convergent findings from autism genetics showing that many other implicated genes function in synapse formation.

What is important about the iPS cells, compared to the information that can be learned from studying mouse cells with MeCP2 knocked out, is that they give a picture of the effects, first, of the specific mutation in this gene in each patient, and second, of the genetic background of each patient, which may modify the effects of the MeCP2 mutation. This gives a far more direct view of the specific effects of each patient’s complete genotype on the development and function of their neurons.

While defects in synapse formation suggest a fundamental role for MeCP2 in neural development, which might imply an irreversible defect, in fact several lines of evidence suggest that the requirement for the function of MeCP2 may be ongoing, in processes of activity-dependent wiring, where neurons within networks strengthen connections based on their patterns of activity. This fits with the apparently normal early development, prior to age two, of girls with Rett syndrome, and also with evidence from mouse models that restoring MeCP2 function in adults can largely reverse the symptoms. These discoveries therefore hold out the promise that intervention in Rett syndrome patients, even in older children, may be effective.

Gage and colleagues tested a couple potential therapies on the neuronal networks derived from Rett syndrome patients and were able to show some degree of rescue of the defects. One of these, the protein insulin-like growth factor-1 (IGF-1), was previously shown to be effective in partially rescuing the defects in MeCP2 mutant mice, most likely by stimulating greater synapse production and compensating for the loss of MeCP2 activity. Clinical trials are now planned to test the efficacy of this approach in patients. Having the cells derived from patients should also greatly facilitate screening for new drugs that can correct the neuronal network defects.

Another paper from the same group, also analysing these cells, revealed a far less expected effect – one that suggests (far more speculatively) the possible involvement of a totally different pathogenic mechanism. One of the functions of the system that methylates DNA is to defend the genome against invaders. Our genome is riddled with parasitic elements – pieces of DNA that can replicate themselves and “jump” around the genome. Fully 45% of our “human” genome is made up of these so-called transposable elements. Most of the copies of these elements are inactive but a subset can generate new copies that will integrate at random into the genome. What has this got to do with Rett syndrome?

Well, MeCP2 is apparently one of the proteins whose job it is to shut down these transposable elements. Gage and colleagues could show that one particular class of these elements, called L1 elements, was far more active in cells derived from Rett syndrome patients. The L1 elements expressed higher levels of the proteins they encode and they generated additional copies of themselves, which were scattered around the genome. Interestingly, this effect seems to be restricted to neurons, presumably because the function of MeCP2 is especially required in that cell-type.

Though highly speculative, this raises the idea that high rates of somatic mutation (somatic meaning it happens in the body, not in the germline and thus will not be inherited), caused by L1 elements jumping around and landing in the middle of genes, may contribute to the severity and also the variability of the phenotype caused by MeCP2 mutations. The alternative is that the L1 transposition has no pathogenic effect but is simply a consequence of the Rett syndrome mutations. Future experiments will be required to tell which of these possibilities is correct.



Marchetto MC, Carromeu C, Acab A, Yu D, Yeo GW, Mu Y, Chen G, Gage FH, & Muotri AR (2010). A model for neural development and treatment of rett syndrome using human induced pluripotent stem cells. Cell, 143 (4), 527-39 PMID: 21074045

Muotri AR, Marchetto MC, Coufal NG, Oefner R, Yeo G, Nakashima K, & Gage FH (2010). L1 retrotransposition in neurons is modulated by MeCP2. Nature, 468 (7322), 443-6 PMID: 21085180