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      Genome editing
      CRISPR girls grow up. How are Lulu, Nana and Amy?

      [Wiki: https://en.wikipedia.org/wiki/CRISPR_gene_editing and https://en.wikipedia.org/wiki/He_Jiankui_affair%5D

      According to rumors they are healthy. But we don’t have enough information, nor technology sophisticated enough, to assess how editing will affect their lives.

      Juggling between no-comments and statements made on condition of anonymity, Nature Biotechnology took stock of the case of the twin girls born in China in the autumn of 2018 and the third CRISPR baby who came to light a few months later as part of the same project. The fictional name assigned to the latter implies that it is another female: Amy . We can continue to call the first two Lulu and Nana , the pseudonyms chosen three years ago by Shenzhen University biophysicist He Jiankui to announce their birth on YouTube. The revelation came after a journalistic scoop and was confirmedby the scientist himself at the International Human Genome Editing Summit in Hong Kong.

      The reconstruction, signed by Vivien Marx, also refers to the information gathered in China by Eben Kirksey, anthropologist at Melbourne’s Alfred Deakin University and author of a book dedicated to the case, entitled “The mutant project”. Apparently, Lulu and Nana were born preterm and needed incubators at an unnamed hospital. When the summit took place on November 28, 2018, a little girl had already been discharged, while her sister had been transferred to the neonatal intensive care unit of another hospital. The other pregnancy, which had been going on for a few months, had been started in the wake of the apparent “success” of the first attempt, despite some dissent in He’s circle. The applause of the Chinese media lasted only a few days, as recounted in the episode dedicated to CRISPR of the podcast ” Reshape – a journey into the medicine of the future ” , the time to understand that the international scientific community almost unanimously condemned the experiment. When Amy came into the world, in an unknown location, the reckless scientist had already been abandoned to his fate: his laboratory had been closed, he fired and kept under surveillance. Then, at the end of 2019, He will be sentenced to three years in prison for abusive practice of the medical profession, so that in the future it will be difficult for him to carry out the project of founding an assisted reproduction clinic with associated genetic editing, as would have been his intentions.

      Experts working on the World Health Organization (WHO) report on human genome editing have repeatedly asked Beijing for access to information gathered by the Chinese committee charged with investigating the case. But the request fell on deaf ears, according to Robin Lovell-Badge, who works at the Francis Crick Institute in London and is a member of the WHO ad hoc panel . Nature Biotechnology , however, reports that DNA extracted from umbilical cord blood and placenta tissue has been sequenced and analyzed. The girls would receive a check-up at birth, at six months and at one year of age. The next exams were scheduled for these days.They likely include standard tests to monitor growth and health, including red and white blood cell counts, hemoglobin, platelets, hematocrit, lipoproteins, and metabolic markers.

      The original monitoring plan devised by He Jiankui should, in theory, continue until the age of 18 , after which it would be up to Lulu, Nana and Amy to give their consent. But it is not known when and how they will be made aware of their status as the first genetically modified humans before they are born, nor what psychological consequences this discovery could have. Meanwhile, liver function tests are expected to be performed at the age of five, while at ten the IQ should be measured. Tests for HIV should also be carried out, because the girls are daughters of HIV-positive fathers and genetic editing was supposed to make them resistant to a hypothetical future infection, by sabotaging the receptor used by the AIDS virus to enter cells.It is unclear whether the plan is going ahead as planned, but the costs of the exams are covered by people close to He . According to various sources, the Chinese authorities have also shown interest in carrying out medical supervision and this additional attention has increased the concern of parents, who do not want to attract suspicion on the girls because they fear a scandal and the consequent social stigma.

      The procedure to which Lulu, Nana and Amy were subjected is the same, but the editing gave different results in the three girls. Lulu shouldn’t be protected from infection, because CRISPR deleted 15 base pairs in one copy of the gene, but left the other intact. Amy, like Lulu, would also have only one edited allele. Nana, on the other hand, could be resistant to HIV, because both copies of the gene for the CCR5 receptor would be altered, albeit in a different way (with more bases in one and fewer bases in the other). At a minimum, therefore, the benefits of treatment appear uncertain. But what can we say about the risk that editing has hit off target , perhaps triggering tumor proliferation?

      The mother of the twins revoked her consent to the amniocentesis, so the only fetal DNA available to Lulu and Nana is that recovered from maternal blood. These genomes were compared with those of mom and dad, looking for any dangerous mutations. In addition, preimplantation genetic diagnosis was performed before transferring the embryos to the uterus. But the scant data He has made public indicate that the genetic modification of the embryos did not occur early enough for all the girls’ cells to inherit uniformly. Well, none of the tests carried out allows us to estimate the potential problems associated with this phenomenon called mosaicism . And actuallythere is no method for estimating the rate of genomic inhomogeneity in an individual while he is still alive . Single cell analyzes can only be performed on easily accessible tissues such as blood. Furthermore, if a few cells are sampled, it may happen that only the unedited ones are taken. And to think that to do things right it would be necessary to test not only Lulu, Nana and Amy, but also their future offspring.

      The most concerned researchers, among those interviewed by Nature Biotechnology , point out that, even where it worked, the CCR5 protein modification is not the same as “delta 32 ,” the naturally occurring deletion in a lucky minority of HIV-immune individuals that He would have liked to reproduce . We do not yet know if these differently altered forms can cause any undesirable effects. Then there is the problem of off-target modifications, which occur when CRISPR cuts sites other than the chosen one. It is not known whether any “off target” edits have been found in Amy’s genome. But He said he found one in an intragenic region of Lulu’s chromosome 1, adding that it shouldn’t have any consequences. But if one has been found through random sampling, it is safe to assume that there may be others . The Harvard geneticist George Church goes against the trend, according to which the rate of off targets induced by editing is below the rate of spontaneous mutations. It would take a certain amount of bad luck for some off-target edit to end up affecting a delicate portion of the genome, such as a tumor suppressor.

      Other fears focus on the risk of macromutations caused by cuts near the chosen site, and therefore “on target” . In fact, human embryos seem particularly predisposed to develop more or less extensive genetic lesions at the editing site , triggered by the welding processes that take place after cutting. Even in natural pregnancies there is a high rate of miscarriages, try to reassure Church. But the orientation of the scientific community as a whole is marked by prudence. While therapeutic editing is demonstrating great potential for correcting genetic defects in the adult cells of diseased individuals , the story of Lulu, Nana and Amy stands as a cautionary tale:using CRISPR on embryos for reproductive purposes still appears to be a gamble with too many unknowns.

      To learn more about how the CRISPR genome editing system works, its discovery, its therapeutic and diagnostic applications, its promises and limitations, listen to the podcast “Reshape – a journey into the medicine of the future” on Spotify (or your favorite platform) and read the illustrated story .

      By: Anna Meldolesi , 06 December 2021

      ***

      Don’t COVID vaccines do the same thing? Aside from injecting all the diseases, the skin rash comes from HIV.

      The only difference is that this Chinese doctor seems really bona fide to me.

      • Un giorno faremo “scudo” a tutte queste malattie… c’è tanto lavoro da fare, anche perché con questo vaccino COVID il mondo non è più un posto sicuro.Per i giovani e per i bambini che nasceranno 🙁

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