Medicine will be brought out of the impasse by microcins and bacteriophages

First, about personalities. About the fact that a professor of Rutgers University (USA) works at two institutes of the Russian Academy of Sciences — molecular Genetics and gene Biology Konstantin Severinov, the general scientific (and not only scientific) public became aware last year after a series of publications in various publications about the attempts of a Russian American to resist the "peculiarities" of the domestic academic system. Of course, some scientists have previously allowed themselves to publicly criticize the RANOV bosses, but perhaps no one has ever acted so consistently and boldly. The publications had an unprecedented resonance and brought Mr. Severinov, a young scientist by Russian standards, "political" popularity.

However, all these are just touches to the portrait, the first, lying on the surface of the association about the activity (and I want to say "adventure") a former graduate of Moscow State University, and now an American professor in Russia. In fact, Konstantin is engaged not only (and not so much) in the struggle for decent working conditions for scientists of the Russian Academy of Sciences, first of all, he studies fundamental issues of modern microbiology.

Together with Konstantin, Natalya Akulenko, a 2008 presidential scholarship in biology, candidate of Biological Sciences, as well as graduate students Timur Kazakov, Ksenia Pugach and Maria Novikova, who revealed to me a terrible secret that after defending their dissertation they would like to continue working in their supervisor's American group.

Now actually about science. Let's make a reservation right away that it is not necessary to directly link fundamental projects, the purpose of which is to find answers to the riddles of nature, with the creation of new medicines. Scientists in Severinov's laboratory do not develop antibiotics as such, but conduct fundamental research, the results of which can ultimately give a much more significant effect for the development of new areas of medicine than a simple fine-tuning and the issuance of a specific new drug so expected by some.

Konstantin Severinov's projects are supported by grants from the US National Institutes of Health, RFBR grants, the academic program "Molecular and Cellular Biology" and the Russian charity foundation "Science for Life Extension".

"Microcins have been introduced to a new method of interferon production"One of the projects, launched in 2000 in the American laboratory of Rutgers University and continued since 2005 at the Institute of Molecular Genetics of the Russian Academy of Sciences, is dedicated to microcins encoded in bacterial genes.

In nature, bacteria use them to fight their own kind. "When there are too many microbes, some of them begin to produce microcins that are safe for themselves, but deadly for their fellows," explains Professor Severinov. — A kind of genocide, during which more adapted and dexterous bacteria clear space for themselves and their descendants to live. Everything is like people."

It would seem that microcins with antibacterial properties should be used in medicine, but the practical use of these substances is hindered by the lack of understanding of the mechanisms of their effect on bacteria. The principles of operation of two microcins were described in detail in Severinov's laboratory. "One of them — microcin C — uses the tactics of the Trojan horse," says Konstantin. — The bacterium takes it for food, absorbs it and begins to break it down, which leads to the release of a deadly substance. Thus, the microbe actually commits an act of suicide." Microcin-like substances created by scientists acting on this principle have already been patented in the USA and Russia. Perhaps in the future they will become the basis for the creation of new classes of antibiotics, but this, as Severinov notes, is already an area of interest in applied science.

The researchers also managed to uncover the "secret" of microcin J, which withstands autoclaving without loss of antibacterial properties, unlike most biologically active substances. "It turned out that its protein chain is literally tied into a strong knot, which explains its unusual physical properties," explains the project manager. — At first we thought that the structure was unique, but as a result of a bioinformatic analysis conducted last year together with the group of the Deputy director of the A.A. Harkevich Institute of Information Transmission Problems of the Russian Academy of Sciences Mikhail Gelfand, we received a whole series of similar substances. Recent publications in a number of leading journals confirm that at least some of them actually exist in real life. However, there is still no information about the biological function of these molecular “nodules" in science. Both in the States and in Russia, my colleagues and I are actively working on this direction, and we are looking forward to receiving information from competitors' laboratories."

Now scientists are busy modifying the genes of the studied microcins in order to give them new, improved properties, thanks to which they could affect resistant forms of bacteria. The obtained substances are screened and the most interesting of them are studied.

In Severinov's laboratory, something is cloned, synthesized and tested every day. In our presence, clones of cells carrying plasmids with a complex of genes for the synthesis of mutant derivatives of microcin C were applied to the surface of agar poured into Petri dishes and containing various test bacteria. After incubation at a temperature of 37 degrees Celsius for several hours, necessary for the formation of a lawn of test cells, clearly visible transparent growth inhibition zones formed around the colonies of microcin-producing cells — the result of the antibacterial action of microcins. Some of these areas were much larger than the zones observed around the unmodified microcin producing cells. These are potential candidates for further study. The subject of the study will also be iterative cycles of additional changes, as a result of which substances with a modified spectrum of antibacterial action can be obtained.

"So, we are experimentally studying the structure and functions of microcin molecules and simultaneously creating libraries of derivatives of these substances, many of which have improved properties and can directly go into the development of promising antibacterial agents or agents for decontamination," says graduate student Timur Kazakov.

In the process of work, unexpected results also happen. Last year, Timur in some mysterious ways left microcins to study cellular proteases, which, in turn, allowed him to find a possible solution to one of the problems relevant to domestic manufacturers of recombinant interferon.

"Interferon produced in Russia will cease to be used in the near future, because after Europe and we will introduce stricter requirements for its chemical composition," explains Konstantin Severinov. — Domestic recombinant interferon, obtained from specially created bacteria, has amino acids that the “natural” human interferon does not have, and which, according to new requirements, must be removed. The obvious solution to the problem is a complete rejection of the “bacterial” interferon, but the level of development of domestic biotechnology does not allow this to be done. Using the knowledge we have gained in the process of studying microcins, it is possible to remove unwanted amino acid residues without completely changing the existing technological platform."

In parallel, scientists at the Institute of Molecular Genetics of the Russian Academy of Sciences are exploring new possibilities for creating microcin-like substances with specified properties (the number of chemical modifications of microcins is limited). In particular, they came to the conclusion that it is possible to obtain extensive "libraries" of microcin variants using standard genetic engineering methods, since microcin peptides are encoded by genes. Presumably, screening of such libraries will lead to the identification of mutant microcins with improved properties and, in the future, to the creation of a replacement for currently used antibiotics.

"A phage fits a bacterium like a key to a lock"The second topic, actively being developed in the laboratory of Konstantin Severinov, is bacteriophages, or bacterial viruses, which were considered as a promising therapeutic agent at the beginning of the last century, but were pushed to the medical "periphery" by synthetic antibiotics.

Perhaps the only place where scientists have never written them off is the G. Eliav Research Institute of Bacteriophages, Microbiology and Virology in Tbilisi, founded in the 20s with the participation of the French bacteriologist Felix d'erell, who, in fact, was the first to describe these viruses. The Institute continues to study phages even today, although it is far from being on a "Soviet" scale.

How to replace antibiotics?
Medicine is gradually becoming almost as helpless as it was before the discovery of penicillin in 1928 by the Scotsman Alexander Fleming and the subsequent boom in antibiotics. Back in the middle of the last century, alarming reports began to appear about cases of dangerous infections in which pathogens remained resistant to seemingly life-saving drugs. Over time, more and more bacteria have discovered their ability to resist synthetic agents. In response, scientists created new, more powerful antibiotics, and gradually became convinced that ... they were coming to a dead end. "We can look back at the era of antibiotics simply as a passing stage in the history of medicine, as an era when huge natural resources were wasted, and microbes turned out to be smarter than people," Ken Harvey, an Australian microbiologist, admitted in one of his articles. In the last few years, scientists in many developed countries have become interested in bacteriophages.

"Phages have obvious advantages over synthetic antibiotics or microcins," Severinov explains. — In the treatment of bacterial infections, they destroy only “enemy” microorganisms, while not affecting the rest of the microbial flora and, therefore, not causing dysbiosis. A phage particle, like a key to a lock, is suitable only for a certain type or even strain of bacteria. At the same time, it not only infects and kills a single microbial cell, but also produces offspring capable of continuing the work of the “parent” as long as phage-sensitive bacteria are present.

At the same time, the scientist admits that practical medicine cannot yet make extensive use of phages, since few characterized phages are known and, in addition, bacteria quickly develop resistance to them, almost as well as to antibiotics.

"We will be able to get the maximum benefit from bacteriophages only after their careful study," Severinov continues. — As viruses created by nature itself to destroy bacteria, they must contain proteins that are toxic to the host cell. We need to find and identify these proteins, and then try to develop an effective way of influencing bacterial cells based on them."

In Severinov's laboratory, phages are considered a very convenient experimental model for studying the work of genes. They allow you to track the process of infection of the cell. "First, the virus attaches to the bacterium, then pierces its shell and injects its DNA inside, and then changes the work of the genes: the genes of the host bacterium cease to act and the genes of the virus are triggered. The process is genetically programmed, it happens quickly and efficiently," says graduate student Ksenia Pugach.

In January of this year, Ksenia conducted an experiment, the results of which opened up the opportunity to clarify the general mechanism of the emergence of resistance of bacterial cells to bacteriophages, unknown at that time. Radioactive phosphorus-32 was needed to continue the work, but the only Russian plant where the coveted substance was produced was closed for several months, and supplies from abroad were not organized, which forced Ksenia to suspend experiments. In August, an article by American and Dutch scientists was published in the journal Science, which described a similar study, however, brought to a qualitatively new level.

"It has become pointless to continue this work," Ksenia says with regret. "Although, in general, we continue to study bacteriophages, and we will try to conduct priority studies and publish the results."

According to Professor Severinov, the sad truth of life is that from the point of view of the organization of laboratory work for a scientist in Russia there are no advantages, but there are a lot of obvious disadvantages. "However, it would be wrong to think that all science is being done or not being done depending on the availability of a device or material," Konstantin believes. — Scientists are people first of all with a head, not with devices. A good researcher can always leave his contribution to science, just in more favorable conditions this contribution will be more significant… After these words, fans of the status quo and vivisection experiments on Russian science will probably be happy - there will always be people who will work anyway, because science is a vocation for them."

Natalia Bykova, STRF.ruPhoto: Dmitry Europin

Portal "Eternal youth" www.vechnayamolodost.ru14.10.2008