Ke-Hui Cui M.D., Ph.D.

Savannah, Georgia, 31405, U.S.A.

Oct. 1 2020

Email: khcui72@hereditics.net

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Preimplantation genetic testing for aneuploidy (PGT-A) is performed by a traumatic embryo biopsy. Now a lot of babies were born after embryo biopsy and they are growing up. Whether embryo biopsy (a major surgery on human embryos) to be safe or not should not depend on simple phenomena: a lot of babies were born, they are healthy,  growing up, study well and running well, etc. The safety question of embryo biopsy should be answered scientifically by research with statistical data accompanying with the control group for comparison. The data of this paper are collected thoroughly from all available papers in the world by internet PubMed, mainly focusing on birth, after birth and growing up with medical and mental testing data. The total available papers are scarce, only about several more than 10 papers could be found and collected in human PGT field since 1989 up to now (over 30 years). None of these papers showed the statistical data to be 100% normal in embryo biopsy group. Most simplest data were birth weight and premature birth, in which no paper reported that the embryo biopsy group was the same as the control group or better than the control group.  They were mainly from some bigger countries with more cases of PGT-A. Higher level of safety research were focusing more on mental and neuropathy research. These research mainly were reported by some smaller countries. Those children with embryo biopsy history were much easier to be watched and organized together locally to perform those deeper research on mental condition. No students' score, immune condition and any other condition such as heart and lung function, physical exercise or physical skill comparison, or fertility and cancer possibility were tested and reported until now. Although some papers seemingly concluded in abstract as "normal" such as in a Isreal paper [Sacks, et al. 2016], the detailed data in the papers did show a lot of contradictory and unsafe results with high significantly different (<0.001). Thus the matchless and groundless "normal" conclusion in the abstract was omitted in this paper and the detailed data in the content of those papers were collected. In these papers of mental research, no any data confirmed that all of the research results of the embryo biopsy group were the same as the control group. All of these papers showed at least one, or two, or more testing items were significantly different. Since blastocyst biopsy (with laser and freezing) on day 5 is harsher procedures than blastomere biopsy  on day 3, the unsafe consequence will be much severe than day 3 biopsy. Also blastocysts are differentiated rather than totipotent as day 3 embryos. Any harm to the embryonic cells will lead to permanent harm to the future babies, children and adults. However, no any safety research results specially comparing between blastocyst biopsy and control group was reported up to now. The author of this webpage got rich experience about: why unsafe results from human embryo biopsy being very difficult to be published is that the results to be too bad to be allowed for publication by journal editor and referees, or even by website hosting services and online search machine companies. They wish to cover up the short coming of PGT and embryo biopsy for the aim of their higher income from more publication for PGT-A. The harmful effect of laser (electromagnetic ray) biopsy was obvious: in Denmark, “compared with spontaneously obtained pregnancies, the risk of placenta previa after PGD was only significant in pregnancies obtained after embryo biopsy with laser but not acid” [Bay, et al., 2016]. In our laboratory, when used laser biopsy with a thermal glass plate, no any human embryos (>30 embryos) could be surviving normally after freezing and thaw cycle.  When replaced the thermal plate with a hole plate, the embryos could recovered significantly better (p<0.01). It confirmed that the laser can be refracted by the thermal glass, plastic dish and water to any direction to harm every cells of our human embryos. The laser scattered to everywhere with only one laser shot was documented with photos demonstrated in the webpage of "Laser Scattered Severely in Human Embryo Biopsy - Harmful". When laser is used, the remained embryonic cells appear morphologically normal under the microscope. However, the microtubules in the cells (i.e. the future nerve fibers after cell differentiation) were obviously observed with cut and broken points by laser with nanotechnique - electron tomography [Laband, et al., 2017]. The severe unsafe results from recent blastocyst biopsy with laser (rather than pregnancy rate) might be the key reason for UK HFEA to put red light on PGT-A in April, 2021.  

The unsafe and negative consequences of embryo biopsy after birth and childhood development are described as “embryo biopsy syndrome” (EB syndrome). In the collected paper data, embryo biopsy was associated with lower rates of live births and higher rates of miscarriage, perinatal death, placenta previa, and premature births with low birth weight. Additionally, children born after PGS showed higher likelihood of major abnormalities, more paramedical care, lower fluency and neurological optimality scores, more problem behavior, poorer cognitive and motor skills, lower levels in stress assessments, lower performance IQ,  lower intensity, significantly higher threshold of response in activity, significantly less moody (negative mood) and significantly less active, etc. Only with new knowledge of Epicytohetics, scientists and physicians, geneticists and lawyers begin to understand the existence of EB syndrome after imperfect embryo biopsy, especially in blastocyst and laser biopsy. EB syndrome and MRT syndrome are both resulted from the damage of cytoplasm, while MRT syndrome usually shows  more severe neuropathy, immune, muscle (including heart muscle), fertility, cancer and other problems.

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In the following international retrospective studies, half of the embryos underwent acid biopsy and half underwent laser biopsy. After 2015, almost all blastocyst biopsy were performed by laser biopsy.

 

In Denmark, “compared with spontaneously obtained pregnancies, the risk of placenta previa after PGD was only significant in pregnancies obtained after embryo biopsy with laser but not acid.” [Bay, et al. 2016]

 

In Spain, a systematic review of 10 trials involving 1,512 women showed that PGS was associated with lower rates of ongoing pregnancies and live births [Checa, et al., 2009].

 

In the Netherlands between 1995 and 2014, 439 pregnancies in 381 women after PGD/PGS resulted in 364 live born children by 311 deliveries. Miscarriage and perinatal death rate = (439 – 311)/439 = 29.2%, which was extremely high. It resulted in 2.5% children with major malformations, 1.4% with minor malformations, 1.1% with chromosomal abnormality, 20% premature births, and <15% with low birth weight [Heijligers, et al., 2018a]. PGS was associated with a significantly lower rate of ongoing pregnancies and live births in women of advanced maternal age [Mastenbroek, et al. 2007]. Other studies have shown that children born after PGS have a 2.5% chance of major abnormalities [Beukers, et al. 2013] and that they receive more paramedical care (for speech, physical, or occupational therapy) than controls [Seggers, et al. 2013]. From a cognitive standpoint, PGS children have been shown to have lower neurologic optimality scores and lower fluency scores compared to controls [Middelburg, et al., 2011, Schendelaar, et al. 2013]. Regarding socio-emotional development, teachers’ scores revealed more externalizing (p = 0.011) and total problem behavior (p = 0.019) in PGD children than in IVF/ICSI children [Heijligers, et al. 2018b].

 

In the UK, PGD/PGS children were more likely to have lower birth weight, at <2500 g (24.5%), compared with naturally conceived controls (1.5%; p < 0.0001), and earlier birth than controls (38.2 ± 2.6 versus 40.0 ± 1.4 weeks; p < 0.0001) [Banerjee, et al., 2008].

 

In 102 two-year old children in Belgium, two PGD/PGS children had major malformations at birth. In PGD/PGS children, about 2/3 underwent a two-blastomere biopsy and these children’s body weights were lower than the controls (p = 0.021) [Desmyttere et al., 2009]. In 581 post-PGD/PGS children (Belgium), the pregnancy rate was extremely low [484 / (1443+1310)] = 17.6%. Moreover, the perinatal death rate was significantly higher than that in the control (ICSI) group [Liebaers, et al. 2010].

 

In Greece, PGD children had increased frequency of poor cognitive and motor skills. In parenting stress assessments, PGD parents reported lower levels of parental stress which were almost twofold difference compared to naturally conceived children (p < 0.01). In addition, the occurrence of defensive response significantly differed between the groups evaluated. Furthermore, PGD parents more often reported lower stress levels in their children as compared to the controls [Thomaidis, et al. 2012].

 

In a study in Israel, 11 (40.7%) PGD children demonstrated mixed hand-eye dominance, while this was found in just 20% of general population. This was double the prevalence of the general population and may reflect how PGD affects individuals’ cognitive functioning. In another study, 12 (44%) PGD children had a significant difference between their verbal IQ and performance IQ (PIQ) scores due to obviously low PIQ scores (compared to 27% in the general population). This finding suggests that subjects may have mixed lateralized cerebral hemisphere dysfunction. Furthermore, PGD children showed significantly lower intensity (p < 0.001) and a significantly higher threshold of response in activity (p = 0.007). They were significantly less moody (negative mood) (p < 0.001) and significantly less active (p < 0.001) [Sacks, et al. 2016]. These phenotypes of EB syndrome show that the invasive biopsy procedure may have affected the developing fetus and children on both medical and neuropsychological levels.

 

In the USA, using national data between the 2005 and 2013 donor oocyte – recipient cycles, it was reported that “pregnancy and live birth rate were significantly lower in preimplantation genetic screening cycles than in control cycles. Live birth rates for preimplantation genetic screening cycles were reduced by 35% (p < .001)” [Barad, et al. 2017]. According to data from the Centers for Disease Control and Prevention (CDC), premature birth rate (34-36 weeks of gestation) in the U.S.A. was significantly increasing between 2015 and 2018 annually (2014: 6.82%; 2015: 6.87%; 2016: 7.09%; 2017: 7.17%; 2018: 7.28%; p < 0.05) [Martin and Osterman, 2018; Hamilton et al., 2019]. In Obstetrics, a higher premature birth rate means that more babies may develop mental retardation. This USA data suggests that a great scale of blastocyst biopsy with laser radiation and cryopreservation since 2015 has been associated with significantly harmful effects on pregnancies and future babies.

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Why will improper embryo biopsy produce unhealthy babies with EB syndrome? Because improper techniques and skills in biopsy may harm the cytoplasm and result in poor differentiation of cytoplasm. Epicytohetics, the study of expression of cytoplasm, explains how cytoplasm damage can lead to abnormal, unhealthy offspring with normal genetics (DNA) but abnormal cell development, abnormal placenta, nerve axon, immune and muscle system, and sperm tail, etc.

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When cytoplasm is damaged by embryo biopsy and laser, it will influence the normal function of cytoskeletons and heredity control system (checkpoint and licensing system) in the cells. It will lead to the time for each cell cycle to be delayed in heredity, i.e. each cell cycle time will be longer than the control [Bar-El et al., 2016]. The baby from the biopsy embryo will finish lesser cell cycles than the control at birth, i.e. the baby contain lesser cell number than the control. Thus lower birth weight in the biopsy group is reasonable. due to containing lesser cell number. It is not a healthy result, because the cell number of the brain of the biopsy group will be lesser than the control group too, which will influence mental ability.

 

Pregnancy maintaining up to full term mainly depends on normal function of the baby and the placenta, especially the placenta to secret high level of estrogen and progesterone. Improper embryo biopsy, especially trophectoderm biopsy, will lead to lesser fetal and placental cell number. When fetus growing further and increasing abdominal pressure further, the blood supply to placenta will drop down. It will lead to estrogen and progesterone production drop down to let delivery happen. Poorer placenta function due to abnormal embryo biopsy technique and skill will get estrogen and progesterone production drop down earlier than the control.  Higher premature birth in biopsy group is the result. That is why premature birth happened earlier up to 34-36 weeks in US since 2015 PGT-A great leap.

 

The cytoplasm (such as cytoskeleton and microtubules) is closely related to differentiation. When microtubules are abnormal, they will lead to increased neuronal branching, shortened neutrites, axons that do not connect to postsynaptic cells, and impaired synapse functioning [Dent and Gertler, 2003; Andersen and Halloran, 2012; Dubey, et al. 2015; Van Beuningen and Hoogenraad. 2016; Kahn and Baas, 2016; Delandre, et al. 2016]. These abnormal microtubules do not return to their normal length, numbers, or spatial positions in the embryonic cells, and such abnormalities can be inherited to the further differentiated cells. Thus, the abnormal microtubules can produce abnormal neuron function [Pessoa-Pureur and Wainer, 2007; Middelburg, et al., 2011; Thomaidis, et al. 2012; Sacks, et al. 2016].

 

Normal functioning of neutrophils, macrophages and lymphocytes in the immune system also depend on the intact cytoskeletal and cytoplasm structures [Alberts, et al. 2015; Stinchcombe, et al. 2015]. Altered cytoskeletal structure will influence normal development of both neural and immune systems simultaneously [Yan-Ming, et al. 2012; Moulding, et al. 2013; Gyorffy, et al. 2016]. Thus, immune problems are another consequence of abnormal cytoplasm, which cannot be diagnosed by any DNA or genetic tests due to the normal DNA and genes [Chen, et al., 2016].

 

In muscles, cytoskeleton actin filaments slide past myosin filaments toward the middle of the muscle unit sarcomere to produce muscle contraction [Cooper, 2000]. The heart is the most heavily worked muscle in the body. Even subtle changes in these cardiac-specific contractile proteins – changes that would not cause any noticeable consequences in other tissues – can cause serious heart disease. It can also result in early heart failure [Alberts, et al. 2015]. Thus disturbance of the cytoskeleton will likely produce severe muscle and heart problems. Stillbirth and perinatal mortality can result from this [Prather, et al. 1989; Checa, et al., 2009; Liebaers, et al. 2010; Heijligers, et al., 2018a].

 

In brief, cells are the basic hereditary and developmental unit of the organs and tissues in the human body. Changes in the cellular cytoskeletons will produce inferior cellular structures and inferior differentiation of all kinds of cells, despite these cells appearing morphologically normal under the microscope. These inferior cells produce inferior organs and systems (such as inferior placenta) with inferior functionality [Bay, et al. 2016], ultimately producing complications such as premature births, babies with low birth weight, and children with low body weights, [Banerjee, et al., 2008; Desmyttere et al., 2009; Heijligers, et al., 2018a; Martin and Osterman, 2018; Hamilton et al., 2019] and different types of neuropathy [Sacks, et al. 2016]. Although these less healthy babies contain normal DNA and genes (i.e. they have normal genetics), they have abnormal cytoskeletons and Epicytohetics. 

 

Embryo biopsy syndrome is a new artificial hereditary disease. It will be inherited to future generations by the abnormal ooplasm mainly through the abnormal oocyte. The reason is: oocyte provides more cytoplasm to the offspring. If the cytoplasm of the sperm is not normal in the man from embryo biopsy group, it will mainly influence the sperm swimming speed due to the abnormal microtubules in the sperm tail. Infertility of the man is foreseeable. It was confirmed in mice experiment.

 

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