A spineless solution - A better way to find novel antibiotics
NEW antibiotics are always welcome. Natural selection means the existing
ones are in constant danger that pathogens will evolve resistance to them. But
winnowing the few chemicals that have antibiotic effects from the myriad that
might do, but don’t, is tedious. So a technique invented recently by Frederick
Ausubel of Harvard University and his colleagues, which should help to speed
things up, is welcome.
Dr Ausubel’s method, the details of which have just been published in ACS Chemical Biology, employs nematode worms of a species called C. elegans as its sacrificial victims. C. elegans is one of the most intensively studied animals on Earth (it was the first to have its genome read completely). It is a mere millimetre long, and can be mass produced to order, so it is ideal for this sort of work.
Dr Ausubel set out to make an automated system that could infect worms with bacteria, treat them with chemical compounds that might have antibiotic effects, and then record the results. The device he has built starts by laying the worms on a “lawn” of pathogenic bacteria for 15 hours and then mixing them with water to create a sort of worm soup. It then places the infected worms into individual enclosures, using a machine called a particle sorter that is able to drop a precise number of worms (in this case 15) into each of 384 tiny wells arrayed on a single plate. These wells have, in turn, each been preloaded with a different chemical that is being tested for possible antibiotic properties. Once in place, the worms are left alone for five days.
Until now, researchers engaging in this sort of work have had to monitor each wellful of worms by eye (assisted by a microscope) to determine whether the inmates were alive or dead. To avoid this time-consuming process, Dr Ausubel and his team exposed their worms to an orange stain once the five days were over. The stain in question enters dead cells easily, but cannot enter living ones. They were thus able to distinguish the quick from the dead by colour, rather than propensity to wriggle.
Moreover, using a stain in this way meant they could automate the process by attaching a camera to the microscope, taking photographs of all 384 wells, and feeding the images into a computer that had been programmed to measure the area of orange in a well and contrast that with the total area occupied by worms. When they compared this automated mechanism for identifying dead worms with manual methods that depended upon human eyes, they found it was every bit as effective.
So far Dr Ausubel and his colleagues have managed to test around 37,000 compounds using their new method, and they have found 28 that have antibiotic properties. Their most exciting discovery is that some of these substances work in completely different ways from existing antibiotics. That means entirely new types of resistance mechanism would have to evolve in order for bacteria to escape their effects.
Mass screening of this sort is not, itself, a new idea in the search for drugs, but extending it so that it can study effects on entire animals rather than just isolated cells should make it even more productive. And worms, unlike, say, white mice, have few sentimental supporters in the outside world.
Source of Information : The Economist 2009-09-05