Cockles and Mussels, Alive, Alive-O!

And so they are across the globe, not forgetting clams, a term that can cover all bivalve molluscs – a huge number of species (over 15,000), all having a two-part, hinged shell. The body inside doesn’t have a backbone, making it soft and edible on a scale of keeping-you-alive to orgasmic, depending on the consumer – oysters and scallops are part of the family.

Bivalves are particularly common on rocky and sandy coasts where they potter happily along, generally burrowing into sediment although some of them, scallops for instance, can swim. By and large their only problem is that humans like to eat them.

Clamming up

However, it gradually emerged in the 1970s that there was another cloud hovering over some of these gastronomic delights. Their commercial importance had drawn attention to the fact that soft-shell clams living along the east coast of North America, together with mussels on the west coast and cockles in Ireland, were dying in large numbers. The cause was an unusual type of cancer in which leukemia-like cells reproduce until they turn the blood milky and the animals die, in effect, from asphyxiation. In soft-shell clams, also known as sand gapers and steamers, the disease has spread over 1,500 km from Chesapeake Bay to Prince Edward Island.

A 2009 study had shown that as the disease progresses there is a rise in the number of blood cells that have abnormally high amounts of DNA (in clams typically four times the normal number of chromosomes – i.e. they’re tetraploid). In parallel with this change the cells make increasing amounts of an enzyme called reverse transcriptase (RT).

That was pretty surprising as RT does what its name suggests: reverses part of the central dogma of molecular biology (DNA makes RNA makes protein) by using RNA as a template to make DNA. RT is usually carried by viruses whose hereditary material is RNA (rather than DNA – so they’re called retroviruses). As part of their life cycle they turn their genomes into DNA that inserts into the host’s genome – which gets reproduced (as RNA) to make more viruses.

But how did RT get into clams? Enter Michael Metzger and Stephen Goff from Columbia University in New York, together with Carol Reinisch and James Sherry from Environment Canada, who began to unravel the mystery.

Jumping genes

Using high throughput sequencing they showed that clam genomes contain stretches of about 5,000 bases that came about when the RNA of a virus was copied into DNA by RT (reverse transcriptase) and then inserted into the host chromosome. Normal clams have from two to ten copies of this ‘repetitive element’ that Metzger & Co dubbed Steamer. That wasn’t too surprising as we have repetitive DNA too – it makes up about half the human genome. Many of these repeated sequences can move around within the genome – they’re often called ‘jumping genes’ – and it’s easy to see how this can happen when RT uses RNA to make DNA that can then pop into new sites in the genome. And you might guess that this process could damage the host DNA in ways that might lead to disease.

A long jump?

It turned out that the diseased clams had suffered massive amplification of Steamer to the extent that they carry 150 to 300 copies of the sequence. So that’s about 30 times as many Steamer DNAs being scattered across the clam genome – but how could that cause the same disease all the way from New York to Prince Edward Island? The answer came from peering into the DNA sequences of the tumour cells: they were virtually identical to each other – but they were different to those of their hosts! Meaning? The damage that led to leukemia, caused by shoe-horning 100s of extra copies of Steamer into clam genomes, only occurred once. And the staggering implication of that finding is that the cancer spread from a single ‘founder’ clam throughout these marine-dwelling molluscs. The resemblance to the way the cancer spreads in Tasmanian devils is striking.

Fishier and fishier

Fast forward to June 2016 and the latest contribution from the Metzger group reporting four more examples of transmissible cancer in bivalves – in mussels from British Columbia, in golden carpet shell clams from the Spanish coast and two forms in cockles.

Each appears to cause the same type of leukemia previously found in clams. The disease appears to be transmitted ‘horizontally’, i.e. by living cancer cells, descended from a single common ancestor, passing directly from one animal to another. Indeed, if you transplant blood cells from infected animals into normal clams they get leukemia.

 Species hopping

All that is quite amazing but the genetic analysis came up with an even more bizarre finding. In the golden carpet shell clams DNA from cancer cells showed no match with normal DNA from this species. It was clearly derived from a different species, which turned out to be the pullet shell clam – a species that, by and large doesn’t get cancer. So they have presumably come up with a way of resisting a cancer that arose in them, whilst at the same time being able to pass live tumour cells on to another species!!clam-transfer-pic

Cancer cell transmission between different species of shellfish. Cancer cells can arise in one species (pullet shell clams) that do not themselves develop leukemia but are able to pass live cells to another species (golden carpet shell clams) that do get leukemia (Metzger et al. 2016).

We have no idea how the cancer cells survive transfer. It seems most likely that they are taken up through the siphons that molluscs use for feeding, respiration, etc. and then somehow get across the walls of the respiratory/digestive systems. In the first step they would have to survive exposure to sea water which contains a lot more salt than cells are happy in. The ‘isotonic’ saline used in drips to infuse patients contains 0.9% salt whereas seawater, with 3.5%, is ‘hypertonic’ – cells put in a hypertonic solution will shrink as water is drawn out of the cell into the surrounding solution. Presumably the cells shrivel up a bit but some at least take this in their stride and recover to reproduce in their new host. Equally obscure is how a species can protect itself from a cancer that it can pass to another species.

These amazing findings throw a different light on the care-free underwater life depicted in Disney’s The Little Mermaid, in which the popular song ‘Under the Sea’ fails to mention floating cancer.

Can this happen to us?!!

Well, not as far as we know. But the fact that the known number of cancers that can be passed from one animal to another has now risen to nine does make you wonder. However, there’s no evidence that it happens in humans in anything like the normal course of events. There are examples of person-to-person transfer, notably during organ transplantation, and there is one recent case of cancerous cells from a tapeworm colonising a human host. But these are very rare, the latter occurring in a patient with a severely weakened immune system, and there is no example of spread beyond two people.

Phew! What a relief! So now we can concentrate on following developments both in Tasmania and beneath the waves in the hope that, not only can we go on satisfying our lust for clam bakes and chowders, but that these incredible creatures will reveal secrets that will benefit mankind.

References

AboElkhair, M. et al. (2009). Reverse transcriptase activity associated with haemic neoplasia in the soft-shell clam Mya arenaria. Diseases of Aquatic Organisms 84, 57-63.

Arriagada, G. et al. (2014). Activation of transcription and retrotransposition of a novel retroelement, Steamer, in neoplastic hemocytes of the mollusk Mya arenaria. PNAS 2014 111 (39) 14175-14180; published ahead of print September 8, 2014, doi:10.1073/pnas.1409945111.

Metzger, M.J. et al. (2015). Horizontal Transmission of Clonal Cancer Cells Causes Leukemia in Soft-Shell Clams. Cell 161, 255–263.

Metzger, M.J. et al. (2016). Widespread transmission of independent cancer lineages within multiple bivalve species. Nature 534, 705–709.

Muehlenbachs, A. et al. (2015). Malignant Transformation of Hymenolepis nana in a Human Host. N Engl J Med 2015; 373:1845-1852.

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Another Peek At The Poor Little Devils

A couple of years ago (July 2014) I wrote a piece called Heir of the Dog that featured Tasmanian devils. The size of a small dog, these iconic little chaps are the largest meat-eating marsupials in the world. I’d run into them at The Lone Pine Koala Sanctuary in Brisbane where they’re keeping company with the dozy, furry tree-climbers as part of a programme to save them – the devils, that is – from extinction by cancer.

animal-fact-guide

A Tasmanian devil. Photo: Animal Fact Guide

1024px-sarcophilus_harrisii_taranna

 

 

 

 

 

 

 

imgresTheir problem comes from their inclination to bite one another, thereby directly passing on living cancer cells (causing devil facial tumour disease – DFTD). At that time the only other known example of transmissible cancer was a rare disease in dogs (canine transmissible venereal tumour – CTVT).

Genetic archaeology

DNA sequencing (i.e. whole genome analysis) had shown that the sexually transmitted dog disease probably arose thousands of years ago in a wolf or East Asian breed of dog and that the descendants of those cells are now present in infected dogs around the world.

The same approach applied to the Tasmanian devil showed that the cancer first arose in a female. Cells derived from that original tumour have subsequently spread through the Tasmanian population, the clone evolving (i.e. genetically diverging) over time. In contrast to the canine disease, DFTD is probably not more than 20 years old. Nevertheless, it spread through the wild population to the extent that the species was listed as endangered in 2008 by the International Union for Conservation of Nature.

Which is why a lot of effort is going into saving them, one approach being a number of breeding programmes in mainland Australia, with the aim of transferring uninfected animals to Tasmania.

One good turn …?

We’re all in favour of saving the little fellows, even if you probably wouldn’t want one as a pet. But, smelly and ferocious as he is, the Tasmanian devil is turning out to be remarkable in ways that suggest they might repay our efforts to keep them going. Things have moved apace down under with Greg Woods, Ruth Pye, Elizabeth Murchison, Andrew Storfer and colleagues from the Universities of Tasmania, Cambridge, Southampton and Washington State making some remarkable discoveries.

Infected animals do indeed develop the most unpleasant, large tumours that are virtually 100% fatal – to the extent that DFTD has wiped out 80% of Tasmanian devils in just 20 years. But some animals survive, even though models of the epidemiology say they shouldn’t. Andrew Storfer’s group asked how they pulled off this trick by looking for genetic changes in almost 300 devils. Quite amazingly, they found that even in a period as short as 20 years there were seven different genes that appeared to have changed (i.e. mutated) in response to selection imposed by the disease. Five of these genes encode proteins known to be associated with cancer risk or the immune system in other mammals, including humans. It seems that the mutations help their immune system to adapt so that it can recognize and destroy tumour cells.

In parallel with those studies, Greg Woods and his team now have a vaccine that looks promising early in trials – in other words a way of boosting natural immunity. We are only just beginning to find ways of giving the human immune system a helping hand – hence the burgeoning field of immunotherapy – so anything that works in another animal might give some useful pointers for us.

sick-tasTasmanian devil facial tumour disease.

This has killed 80% of the wild Australian animals in just a few decades.

Photograph: Menna Jones.

As if that wasn’t enough, a second strain of cancer has been found in a small group of male Tasmanian devils. It causes fatal facial tumours that look much the same as the first DFTD. However, it has a completely different genetic cause – so different in fact that it carries a Y chromosome, clear indication that the two forms of the disease arose by quite distinct mechanisms – which makes this marsupial the only species known to be affected by two types of transmissible of cancer.

Milk and human kindness

On top of all that some brave souls at Sydney University, Emma Peel and Menna Jones, decided in that way that scientists do, to collect some milk from the ferocious furries, just to see if it was interesting. Astonishingly the marsupial milk contained small proteins (peptides) that could kill a variety of bugs. They’re called cathelicidins and one of the things they can target is methicillin-resistant Staphylococcus aureus – MRSA – one of the dreaded ‘superbugs’ that are resistant to penicillin and other antibiotics. It’s not clear whether these peptides help to protect the devils from cancer but if that’s how turns out it might be incredibly important for us. As for their antibiotic potential, well, as it’s predicted that by 2050 superbugs will be killing one of us every three seconds you could say that opportunity beckons.

So that’s all incredibly exciting – and not just for the Tassie devils. ­ But another reason for returning to this story is that the devils have recently been joined by another example of extraordinary cancer transmission – and this one comes from the last place on the planet that you’d look for it ….

References

Murchison, E.P. et al. (2012). Genome Sequencing and Analysis of the Tasmanian Devil and Its Transmissible Cancer. Cell 148, 780–791.

Pye, R.J. et al. (2016). A second transmissible cancer in Tasmanian devils. Proceedings of the National Academy of Sciences USA, 113, 374–379.

Epstein, B. et al. (2016). Rapid evolutionary response to a transmissible cancer in Tasmanian devils. Nature Communications 7, Article number: 12684: doi:10.1038/ncomms12684.

Peel, E. et al. (2016). Cathelicidins in the Tasmanian devil (Sarcophilus harrisii). Scientific Reports 6, Article number: 35019. doi:10.1038/srep35019.