New Nobel Prize winner: She saved all mankind with forty years of counterattack and cultivated an Olympic champion

Leaving Hungary from scratch, in order to succeed, we must work tirelessly for our lives, our survival and our success. Congratulations on becoming the 2023 Nobel Prize in Physiology or Medicine!

Written by | Guo Xiaoqiang

Original title: "New Nobel Prize winner: middle-aged unemployed, suffering from cancer, she spent forty years to save all mankind, and also cultivated an Olympic champion"

Since December 2020, the United Kingdom, Canada, the United States and other countries have successively approved the emergency use of the new crown mRNA vaccine jointly developed by Pfizer and Germany's BioNTech, and people finally see hope to end the global new crown epidemic.

Compared with other vaccines, mRNA vaccines have many advantages: first, they are highly safe, mRNA is not infectious, does not have to be integrated into the genome, and its life in vivo can be achieved by in vitro operations as needed (to avoid the harm caused by long-term retention); The second is good immunity, the current mRNA vaccine has been relatively stable, can be efficiently expressed in the cell to produce proteins, and initiate an efficient immune response response; Third, mass production is rapid, and mRNA in vitro preparation technology is very mature, and a large number of vaccines can be prepared quickly and conveniently as needed.

As the world's eyes focus on the new crown mRNA vaccine, the key scientists behind this technology that were once unknown have also surfaced and been interviewed by major media. She is Katalin Karikó, a 65-year-old biochemist from Hungary.

Carrico works from his Pennsylvania home. from STAT

There is no turning back

Calrico was born on January 17, 1955, in a cottage with a sawdust stove in the eastern Hungarian town of Kisújszállás. Scrutinizing the pigs her father slaughtered every day was her science initiation lesson.

In 1973, Kaliko was admitted to the prestigious Hungarian University of Szeged and chose science without hesitation. In college, she first heard about messenger RNA (mRNA), which carries genetic information in DNA, directs protein synthesis, and acts as a "messenger." Karico became intrigued by this amazing molecule. In 1978, she chose to pursue a Ph.D., focusing on the use of mRNA.

In the seventies of the 20th century, genetic engineering was born, and soon the concept of gene therapy was born, but these operations targeted DNA, and Karico thought that mRNA was more promising. After graduating, she chose to enter the Institute of Biophysics at the Biological Research Centre (Szeged) of the Hungarian Academy of Sciences. At that time, many people went to the United States to study after graduating with a doctorate, but Carrico was not impressed, she believed that her wishes could also be realized in China. Unfortunately, Carico's good wishes were dashed in 1985, and she was fired from the unit.

Years later, Mr. Kaliko said in an interview that if she remained in Hungary, she would likely become a complaining, mediocre researcher. With nowhere to go back, Carrico had to start looking for work again. At first, she wanted to find a position in Europe, but in the end, she had to travel to Philadelphia, across the Atlantic. There, Temple University offered her a postdoctoral position.

Without a mobile phone or credit card, the couple took their two-year-old daughter and set foot in a foreign country. The government was not allowed to exchange more than $100 in cash, so they sold their car on the black market and smuggled £900 out of the country in their daughter's teddy bear. "We have no turning back," Carrico said. We were there without relatives. ”

In 1985, Carrico restarted his scientific research at Temple University. Unfortunately, the first stop didn't go well. Four years later, she had a conflict with her mentor, mainly because of their different views on mRNA. Like many researchers at the time, mentors were not optimistic about mRNA research. In 1990, Carrico joined the University of Pennsylvania. At this time, a recent development further strengthened her determination to develop mRNA applications.

Maybe I'm not good enough, not smart enough

In 1990, a team of researchers at the University of Wisconsin injected mRNA into mice for the first time (doi: 10.1126/science.1690918) and detected corresponding protein expression; two years later, another team further demonstrated in rats that the proteins expressed by the mRNA injected in vitro were also physiologically active. If these two results are true, it means that mRNA, which uses key proteins of pathogens, also produces viral proteins and stimulates an immune response that can play the role of vaccine.

This logical reasoning is readily available, but many scientists are not optimistic about it. Because of the practical problems of doing so, vaccines with mRNA have at least three major drawbacks: poor stability (a problem that still exists), inefficiency in the body, and stimulation of the body's innate immune system (causing a severe inflammatory response that leads to the immediate death of the animal). In the eyes of many scientists, these difficulties are insurmountable scientific gaps, especially the third flaw, which may eventually be difficult to resolve. This kind of thankless thing is naturally not a few people are willing to do, and the traditional vaccine preparation strategy is enough, so why bother to stay close?

"Mainstream perceptions" are bound to influence the development of an area. Many of the biggest bulls in mRNA research have shunned and no longer mentioned the use of mRNA as a vaccine, and the resistance encountered by unknown recruits in the field can be imagined.

The year he entered the University of Pennsylvania, Carrico submitted a grant application to try to develop a vaccine using mRNA. In such a mainstream context, the application failed. However, unexpectedly, in the following years, every year of application, every year was rejected, and for eight years could not apply for a fund for this topic. She recalled, "I was writing about funds, writing funds, writing funds, and every time I was called back, called, called. "You have a thousand tricks, I have certain rules; If you say anything, I won't give you the fund. This idea, which does not seem to be very "contrary" today, is rejected by fellow experts. Avram Hershko, winner of the 2004 Nobel Prize in Chemistry, argued that experts always stick to the rules and that many ideas are not worth accepting (because ubiquitin remains active after heating, they conclude that ubiquitin cannot be a protein).

The bosses finally couldn't stand it. In 1995, his sixth year at Penn, Carrico was demoted and his salary cut. She recalled that she had just made some important discoveries, and the school threw her out of the lab and arranged a small room next to the animal room for her to do experiments. To make matters worse, she was diagnosed with cancer and needed two surgeries, while her husband had to stay in Hungary due to visa problems and could not return to the United States for six months. She had to take care of her children while receiving treatment.

Ordinary people have this experience and have long left academia, but Carrico survived: "I want to go somewhere else and study something else." I also thought maybe I wasn't good enough, not smart enough. I tried to convince myself that everything was ready, I just needed to make the experiment more beautiful. ”

Fortunately, Carrico eventually recovered and continued his own experiments. Due to the constraints of all parties, it can be difficult to get things done. She couldn't afford to subscribe to a magazine, and she had to go to photocopy in order to see the latest papers. During a photocopying in 1997, Karico befriended Drew Weissman, an immunologist who had just arrived at Penn. Weisman was intrigued by Kaliko's idea and decided to fund her to continue her research, and her project officially became the "Weisman Kaliko Project." Carrico's situation at that time can be said to have fallen to the freezing point, and the pay is lower than that of the technicians, and Weissman's help can be described as a blessing, not only financial support, but also important spiritual encouragement.

The three-peak loop turns, and then makes waves

Carico's research gradually improved. In 1998, the long-awaited fund was finally approved, and although it was only a measly $100,000, it was at least a good start. The following year, another $1 million was awarded. After discussion, Carrico and Weissman agreed that the safety of mRNA applications needed to be addressed first, that is, to understand why mRNA induces an inflammatory response in the body.

In the nineties of the last century, the elucidation of the innate immune mechanism expanded people's understanding of the immune system. In 1998, American immunologist Bruce Beutler discovered that there is a Toll-like receptor (TLR) family on the surface of immune cells such as dendritic cells, which can recognize bacterial components (such as lipopolysaccharides), and the combination of the two will activate and initiate an innate immune response, and Butler also shared the 2011 Nobel Prize in Physiology or Medicine for this discovery.

Carrico speculates that mRNA injections into animals induce inflammation, possibly because they are recognized by TLR molecules. In order to verify the correctness of his hypothesis, Kaliko first established an in vitro system to simulate the inflammatory response, and applied synthetic mRNA to directly treat cells, which indeed activated the immune response and released a large number of immune factors. Further studies have found that multiple TLR molecules (including TLR7, 8, etc.) can indeed recognize in vitro injected mRNA.

In 2004, Carrico completed a pivotal experiment. She extracted mRNA directly from mammals and bacteria and treated cells with them, and found that mammalian mRNA basically does not activate the immune response (except mitochondrial mRNA), while bacterial mRNA induces the release of cytokines, which indicates that the cause of the immune response is not the mRNA itself, but its structural differences. At the time, it was known that mammalian mRNA had extensive base modification, which was not usually present in prokaryotes such as bacteria (similar to mRNA synthesized in vitro). As a result, Carrico also modified the mRNA synthesized in vitro, resulting in a greatly weakened immune response (later animal experiments also proved that the modified mRNA no longer produces a serious inflammatory response). In fact, mammals recognize unmodified mRNA (foreign components), but the ability to turn a blind eye to modified mRNA is precisely the basic feature of the immune system - distinguishing "non-self" and the body's protection of itself. This finding means that the safety of mRNA in vivo application has been effectively addressed (through in vitro base modification).

Kaliko further found that mRNA synthesized in vitro often contaminates a certain amount of double-stranded RNA, which also triggers an immune response, so she purifies the originally synthesized RNA and removes the double-stranded RNA. On the one hand, this operation reduces the occurrence of inflammation, and more importantly, greatly increases the protein production efficiency of mRNA in vivo, thereby solving the problem of inefficiency in the application of mRNA. Carrico has published more than 70 papers, most of which focus on the improvement and improvement of mRNA in vitro preparation methods and solve many problems faced in practical applications.

In 2006, Carrico and Weissman applied for the first mRNA-related patent - the preparation and application of mRNA containing modified nucleotides, mainly involving mRNA (patent number: US 8278036) that is non-immunogenic and contains nucleotide modifications. To date, she has held more than a dozen patents, all of which revolve around the improvement, practical operation and application of mRNA preparation methods. That year, she co-founded a biotechnology company, RNARx, to try to develop mRNA drugs (mainly EPO mRNA for the treatment of anemia), but the company eventually closed 7 years later. The mRNA boom that Carrico had hoped for did not occur, and the market was not enthusiastic about the research, so it was less noticed.

In 2010, the turnaround came again. Derrick Rossi, who is a postdoc at Stanford University, discovered Carrico's article and was acutely aware of the great potential of this method. He founded a biotechnology company, known as Moderna, to develop vaccines and drugs using mRNA. At the same time, Carrico also transferred his technology to BioNTech, an emerging German biotech company. At the time, BioNTech was still living on the campus of Mainz University in Germany, without even having a company website.

In 2013, Carrico had another unpleasant encounter with the University of Pennsylvania, refusing to reinstate her 1995 salary cut and disagreeing with her over licensing intellectual property (Penn sold the intellectual property to another company). Eventually, Carrico chose to resign to join BioNTech as Senior Vice President. The school was extremely harsh on Carrico, saying that BioNTech was a small, little-known company that didn't even have a website, suggesting that Carrico's choice was worthless.

As mRNA technology is further improved in its application, the two companies are getting closer to true market success. In 2017, Moderna began developing an mRNA vaccine for Zika virus; In 2018, BioNTech partnered with Pfizer to develop an influenza mRNA vaccine, trying to move from the lab to the application. However, the market still does not buy it, investors are not optimistic about the prospect of mRNA vaccine application, and the two companies can only "get by hard".

During these silent and bitter years of research, her daughter Zsuzsanna Francia became famous even earlier than Carrico . Perhaps inheriting her mother's perseverance, Zusana won back-to-back rowing competitions at the 2008 Beijing Olympics and the 2012 London Olympics.

At the 2012 London Olympics, the Carricos congratulated their daughter on winning the Olympic gold medal. Photo courtesy of Katalin Kariko

IV. Perfect Redemption

At the beginning of 2020, the new crown pneumonia broke out and the new crown virus spread around the world.

On January 11, Zhang Yongzhen's research team from the Chinese Center for Disease Control and Prevention published the whole genome sequence of the new coronavirus on the virology website (virological.org).

As soon as the sequence was made public, pharmaceutical companies in Europe and the United States began to study the sequence to be used in mRNA vaccines.

On January 13, the sequence was determined and Moderna began making mRNA.

We all know what happened later.

In the global multi-country new crown vaccine development competition, the advantages of mRNA vaccines (short development time) are fully reflected, and on the basis of obtaining the new crown virus spike protein (S) mRNA information, the steps of design, preparation, animal experiments, and clinical experiments are quickly opened. On November 9, Pfizer and BioNTech jointly announced that based on the results of phase III. clinical trials, the new crown vaccine mRNA BNT162b2 developed by it has an effective rate of more than 90% (the final data shows that the effective rate can reach 95%); A week later, Moderna announced that its mRNA vaccine, mRNA-1273, was also close to 95 percent effective.

When Carrico heard the exciting results of BioNTech's Phase III clinical trial, her first reaction was: "Saved!" I inhaled desperately, I was so excited, I was so afraid that I would die..." The heart that had been hanging for a long time could finally get some rest. Carrico hopes that mRNA vaccines will play an important role in subsequent prevention of new coronary pneumonia, and that mRNA technology will be widely used in the treatment of more diseases.

Rossi, now a professor at Harvard's stem cell institute, believes that if mRNA vaccines finally play a key role in the coronavirus pandemic, Carrico and Weisman definitely deserve the Nobel Prize in Chemistry.

That's more than forty years since Carrico first began studying mRNA, and fifteen years after her key technological breakthrough.

Main references

1. Karikó K, Buckstein M, Ni H,et al. Suppression of RNA recognition by Toll-like receptors: the impact of nucleoside modification and the evolutionary origin of RNA. Immunity. 2005,23(2):165-175.

2. Karikó K, Muramatsu H, Ludwig J, et al. Generating the optimal mRNA for therapy: HPLC purification eliminates immune activation and improves translation of nucleoside-modified, protein-encoding mRNA. Nucleic Acids Res, 2011,39(21):e142.

3. Sahin U, Karikó K, Türeci Ö. mRNA-based therapeutics--developing a new class of drugs. Nat Rev Drug Discov. 2014,13(10):759-780.

4. Pardi N, Hogan MJ, Porter FW, Weissman D. mRNA vaccines - a new era in vaccinology. Nat Rev Drug Discov. 2018,17(4):261-279.

5. Wolff JA, Malone RW, Williams P, et al. Direct gene transfer into mouse muscle in vivo. Science. 1990, 247: (4949 Pt 1)1465–1468.

6. Jirikowski GF, Sanna PP, Maciejewski-Lenoir D, et al. Reversal of diabetes insipidus in Brattleboro rats: intrahypothalamic injection of vasopressin mRNA. Science. 1992, 255 (5047): 996–998.

7. COX D. How mRNA went from a scientific backwater to a pandemic crusher. ( https://www.wired.co.uk/article/mrna-coronavirus-vaccine-pfizer-biontech)

8. A TYPICAL HUNGARIAN STORY: KATALIN KARIKÓ （https://hungarianspectrum.org/2020/11/22/a-typical-hungarian-story-katalin-kariko/）

9. BioNTech scientist Katalin Karikó risked her career to develop mRNA vaccines. Americans will start getting her coronavirus shot on Monday.

（https://www.businessinsider.com/mrna-vaccine-pfizer-moderna-coronavirus-2020-12）

10. The story of mRNA: How a once-dismissed idea became a leading technology in the Covid vaccine race（https://www.statnews.com/2020/11/10/the-story-of-mrna-how-a-once-dismissed-idea-became-a-leading-technology-in-the-covid-vaccine-race/）

11. 'Redemption': How a scientist's unwavering belief in mRNA gave the world a Covid-19 vaccine

（https://www.telegraph.co.uk/global-health/science-and-disease/redemption-one-scientists-unwavering-belief-mrna-gave-world/）