The
matrix
Secret networks could be the key to life – and death.
Anil Ananthaswamy investigates
●
HAD Amin Rustom not messed up,
he would not have stumbled upon one
of the biggest discoveries in biology of
recent times. It all began in 2000, when he saw
something strange under his microscope. A
very long, thin tube had formed between two
of the rat cells that he was studying. It looked
like nothing he had ever seen before.
His supervisor, Hans-Hermann Gerdes,
asked him to repeat the experiment. Rustom
did, and saw nothing unusual. When Gerdes
grilled him, Rustom admitted that the first
time around he had not followed the standard
protocol of swapping the liquid in which the
cells were growing between observations.
Gerdes made him redo the experiment,
mistakes and all, and there they were again:
long, delicate connections between cells. This
was something new – a previously unknown
way in which animal cells can communicate
with each other.
Gerdes and Rustom, then at Heidelberg
University in Germany, called the connections
tunnelling nanotubes. Aware that they might
be onto something significant, the duo
slogged away to produce convincing evidence
and eventually published a landmark paper in
2004 (Science, vol 303, p 1007).
At the time, it was not clear whether
these structures were anything more than a
curiosity seen only in peculiar circumstances.
Since their pioneering paper appeared,
however, other groups have started finding
nanotubes in all sorts of places, from nerve
cells to heart cells. And far from being a mere
curiosity, they seem to play a major role
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in anything from how our immune system
responds to attacks to how damaged muscle
is repaired after a heart attack.
They can also be hijacked: nanotubes may
provide HIV with a network of secret tunnels
that allow it to evade the immune system,
while some cancers could be using nanotubes
to subvert chemotherapy. Simply put,
tunnelling nanotubes appear to be everywhere,
in sickness and in health. “The field is very
hot,” says Gerdes, now at the University
of Bergen in Norway.
It has long been known that the interiors
of neighbouring plant cells are sometimes
directly connected by a network of nanotubular
connections called plasmodesmata. However,
nothing like them had ever been seen in
animals. Animal cells were thought to
communicate almost entirely by releasing
chemicals that can be detected by receptors
on the surface of other cells. This kind of
communication can be very specific – nerve
cells can extend over a metre to make
connections with other cells – but it does
not involve direct connections between
the interiors of cells.
The closest animal equivalents to
plasmodesmata were thought to be gap
junctions, which are like hollow rivets
joining the membranes of adjacent cells.
A channel through the middle of each gap
junction directly connects the cell interiors,
but the channel is very narrow – just
0.5 to 2 nanometres wide – and so only allows
ions and small molecules to pass from one
cell to another.
Nanotubes are something different. They
are 50 to 200 nanometres thick, which is more
than wide enough to allow proteins to pass
through. What’s more, they can span distances
of several cell diameters, wiggling around
obstacles to connect the insides of two cells
some distance apart. “This gives the organism
a new way to communicate very selectively
over long range,” says Gerdes.
Travelling by tube
Soon after they first saw nanotubes in rat
cells, he and Rustom saw them forming
between human kidney cells too. Using
video microscopy, they watched adjacent
cells reach out to each other with antenna-like
projections, establish contact and then build
the tubular connections. The connections
were not just between pairs of cells. Cells
can send out several nanotubes, forming
an intricate and transient network of linked
cells lasting anything from minutes to hours.
Using fluorescent proteins, the team also
discovered that relatively large cellular
structures, or organelles, could move from
one cell to another through the nanotubes.
The first clue to how membrane
nanotubes, as some researchers prefer to
call them, might be used by cells came from
the US. Simon Watkins of the University of
Pittsburgh, Pennsylvania, and his colleagues
were studying dendritic cells, the sentinels
for the immune system. When a dendritic
cell detects an invader, it gets ready to sound
the alarm. One sign of this activation is
15 November 2008 | NewScientist | 43
 It pays to network
The discovery of tunnelling
nanotubes (see main story) has
led to speculation about just how
far their influence extends. Some
think they might play a crucial
role in development.
One of the key outstanding
questions in biology has to do
with how cells communicate with
each other as an embryo develops.
We know that some cells release
molecules called morphogens,
which diffuse through the
intercellular medium. The
resulting concentration gradient
tells cells where they are in the
embryo and they can then develop
accordingly. But this does not
explain everything.
For instance, two very distant
cells can show similar patterns
of gene expression while other
cells nearby do not. If a morphogen
gradient is responsible, then surely
the cells closer to the one releasing
the morphogen should also
be responding.
Such observations could
be explained if morphogens are
distributed directly to certain cells
through a network of nanotubes,
speculates Hans-Hermann Gerdes
of the University of Bergen in
Norway. “It would be much more
appealing for nature to have this
direct line,” he says.
Plants cells are known to
use their version of nanotubes,
called plasmodesmata, to transfer
microRNAs that influence gene
activity from one cell to another
during embryogenesis. “These
microRNAs lead to certain
expression patterns in adjacent
and also very remote cells,” says
Gerdes. Mammalian cells could
also be doing the same.
Even more speculative is
the idea that the development
of organs could be influenced
by tunnelling nanotubes. It is
not well understood how organs
“know” how big they should
get, or the exact shape to take.
“These are all open questions,”
says Gerdes. He speculates that
if the cells in a given organ were
connected by nanotubes, then
these connections could help
establish a feedback mechanism
that provides the necessary
information for the organ to
grow to the right size and shape.

a change in calcium levels in the cell.
While Watkins was poking a dendritic
cell with a micro-needle filled with bacterial
toxins, he noticed a calcium fluctuation in a
dendritic cell far away from the one that was
touched. “Wow, that’s pretty cool,” thought
Watkins. Information about the toxins was
somehow being passed from the cell being
poked to a distant cell. Nothing in his
experience could explain the phenomenon.
When Watkins dived into the literature,
he discovered Gerdes’s paper. His team then
took another look at the dendritic cells. Sure
enough, they found the cells were connected
by a network of tunnelling nanotubes.
Watkins thinks that the dendritic cells
could be using nanotubes to recruit other
cells. Conventional wisdom says that once
a dendritic cell is activated, it migrates to
the lymph nodes to alert the immune system.
Sometimes, it might have to travel from the
tip of one’s finger to the armpit – a long and
perilous journey that could result in failure.
But if a dendritic cell first recruits other
sentinels, and all of them march towards
the lymph nodes simultaneously, there is
much less chance of the message being lost.
“It allows you to amplify the response,”
says Watkins. “That’s all hypothesis really.
We have to prove it.”
Meanwhile, Stefanie Dimmeler of the
University of Frankfurt in Germany and
her team have been studying how so-called
progenitor cells can be transformed into heart
muscle cells. In mice at least, progenitor cells
injected after heart attacks seem to turn into
new heart muscle cells, replacing dead tissue.
When Dimmeler’s team mixed heart
muscle and progenitor cells in a dish, they
found that the two populations established
connections via nanotubes. They even
observed the transfer of organelles such as
mitochondria (Circulation Research, vol 96,
p 1039). Dimmeler suspects that the transfer
via nanotubes of signalling molecules and
proteins called transcription factors helps
to transform the progenitor cells.
The best way to prove this would be to
show that without nanotubes, progenitor cells
mixed with heart muscle cells do not turn into
heart muscle cells as well. The trouble is that
no one has yet found a way of destroying
nanotubes without also damaging the cells
that they are attached to.
While the evidence for the normal roles of
tunnelling nanotubes remains circumstantial,
it is becoming clear that the networks they
form can be hijacked. In a study published
earlier this year, Daniel Davis’s team at
Imperial College London infected some
immune cells with an HIV modified to express
a fluorescent protein, and mixed the infected
cells with healthy ones. “We could literally see
clumps of that protein moving from the
infected to the uninfected cell, along the
strand of the nanotube,” says Davis. This
strongly suggests that the infection may
spread from cell to cell in this way (Nature
Cell Biology, vol 10, p 211).
This might help to explain why some
HIV-infected people, though they carry
antibodies to the virus, cannot seem to
get rid of it. “Somehow viruses are avoiding
immune-system recognition, and one way
they could do that is if they could get from
one cell to another through direct cell
contact,” says Davis.
Another infectious agent that could
spread via tunnelling nanotubes are prions,
the cause of mad cow disease. “One of the key
unresolved issues in prion-disease research
is how prions pass from one cell to another,
as they have to do to spread throughout the
nervous system and cause disease,” says
Byron Caughey of the US National Institutes
of Health in Hamilton, Montana.
The prion connection?
Caughey’s team has already uncovered
one mechanism unrelated to tunnelling
nanotubes by which certain prions spread, but
it is probably not the only way. “At this point,
we are very intrigued by the possibility that
tunnelling nanotubes are partly responsible
for the spread of prion infectivity from cell to
cell,” he says. “Once you know the mechanism
of cell-to-cell transfer, hopefully it opens up
new therapeutic targets.”
Nanotubes may also play a role in tumours
becoming resistant to chemotherapy. Much
of this resistance is due to a class of proteins
called ABC transporters, which pump the
anti-cancer drugs out of cells. It was recently
discovered that tumour cells without these
proteins can acquire them from other
tumour cells. But how?
Prostate cancers cells have recently

44 | NewScientist | 15 November 2008 www.newscientist.com


PAUL MCMENAMIN/UWA
been spotted swapping material via a
network of tunnelling nanotubes. It is possible
to imagine cancer cells using tunnelling
nanotubes to exchange ABC transporters
and spread drug resistance throughout the
tumour, says Gerdes. “If this were to be true,
and if I could find a drug which inhibits the
growth of these nanotubes, I could reduce
the resistance to chemotherapy.”
That is easier said than done, though,
because so little is known about these
nanotubes. They seem to come in many
shapes and sizes, of varying thickness and
lengths, and differ from cell type to cell type.
Precisely how they function is not yet clear.
There are some hints, however. Gerdes’s team,
for instance, discovered that the nanotubes
they studied contained myosin Va, a type
of motor protein. Elsewhere in cells, objects
attached to myosins move along tracks made
of proteins called actins, so Gerdes thinks that
this kind of process might help move things
through nanotubes.
Characterising exactly what these
nanotubes are made of will be crucial.
Gerdes’s team, for instance, is trying to find
proteins that are specific to nanotubes. Once
identified, such proteins can be tagged with
fluorescent markers, making it much easier
to see nanotubes. Such work could also make
it possible to destroy or manipulate these
structures, and thus provide solid proof
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“It is a previously
unknown way in which
cells can communicate
over a distance”
of their importance – and a lot of researchers
still need convincing.
If tunnelling nanotubes really are
ubiquitous, the critics ask, how come they
managed to evade discovery until so recently?
And why have they only been seen in cells
grown outside the body?
There may be several reasons why
nanotubes have eluded notice for so long.
For starters, they are extremely fragile:
merely shaking a dish of cells or changing
the medium – as Rustom failed to do – can
rupture these tubes, as can certain chemicals
used to fix cells for observation, including
those used with electron microscopes. Even
prolonged exposure to light can destroy
them. (This extraordinary susceptibility to
chemicals and light may one day provide
a means to selectively destroy nanotubes.)
In addition, when biologists observe cells
in culture, they usually focus on the bottom
of the dish or slide, where delicate structures
such as nanotubes will be obscured by debris.
Finally, although nanotubes are elusive, many
researchers have spotted them over the years
without realising it.
At the University of Western Australia
in Crawley, for example, Paul McMenamin’s
team has been studying dendritic cells in
mouse corneas. McMenamin’s graduate
student Holly Chinnery kept seeing
something unusual. “She kept noticing
these cells with big, long processes,” says
McMenamin. “She’d show me the pictures,
and I’d say, ‘Gosh, I haven’t seen anything like
that before’.” And so it went until a colleague
told Chinnery about nanotubes. “That
immediately set us off,” says McMenamin.
“We realised that we had the first evidence
of them in vivo.”
Their work, published in May, shows
that nanotubes are not just an artefact of
the methods used to grow cells in culture,
as some have suggested. And what they have
seen is spectacular: some of the longest
tunnelling nanotubes ever observed, more than
300 micrometres long, connecting dendritic
cells in the cornea (The Journal of Immunology,
vol 180, p 5779). “We can see them their whole
course, spindling all the way through the
cornea,” says McMenamin. “It’s fantastic.”
“I’ll bet you that within weeks to months,
people will start noticing them in other
tissues. It’s just a case of how you look,”
he adds. “You’ve got to know what you are
looking for. It’s a bit like being a good bird-
watcher. A hundred people will see a flock of
seagulls, and it’s only a very good bird-watcher
who will spot this one tern flying in that flock.”
Gerdes, meanwhile, continues to marvel
at what is unravelling before his very eyes.
“Whatever one can think of has been done
by nature,” he says. “It is unbelievable what
the cell is able to do.” ●
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