www.cogen2018.cme-congresses.com

Professor Gerald Schatten

University of Pittsburgh School of Medicine gschatten@pdc.magee.edu

Are we ready for genome editing in

human embryos FOR CLINICAL

PURPOSES?

Nothing to declare

www.cogen2018.cme-congresses.com

Professor Gerald Schatten

University of Pittsburgh School of Medicine gschatten@pdc.magee.edu

Are we ready for genome editing in

human embryos FOR CLINICAL

PURPOSES?

Maybe not yet, but it’s inevitable…

Nothing to declare

One particularly controversial application of this powerful gene editing technology is the possibility of driving certain species to extinction – such as the most lethal animal on Earth, the malaria-causing Anopheles gambiae mosquito. This is, as far as scientists can tell, actually possible, and some serious players like the Bill and Melinda Gates Foundation are already investing in the project. (The BMGF funds The Conversation Africa.)

Yang’s genetically modified mushrooms were deemed exempt from current USDA regulation.

Fig. 2 Dystrophin correction after intramuscular delivery of AAV9-encoded gene editing components.

Leonela Amoasii et al. Science 2018;362:86-91

Published by AAAS

Article

Live birth derived from oocyte spindle transfer to prevent mitochondrial disease John Zhang a,b,*, Hui Liu b, Shiyu Luo c, Zhuo Lu b, Alejandro Chávez-Badiola a, Zitao Liu b, Mingxue Yang b, Zaher Merhi d, Sherman J Silber e, Santiago Munné f, Michalis

Konstandinidis f, Dagan Wells f, Jian J Tan g, Taosheng Huang c,*

a New Hope Fertility Center, Punto Sao Paulo, Lobby Corporativo, Américas 1545 Providencia, Guadalajara, Mexico b New Hope Fertility Center, 4 Columbus Circle, New York, NY 10019, USA c Division of Human Genetics, Cincinnati Children’s Hospital Medical Center, OH 45229, USA d Department of Obstetrics and Gynecology, Division of Reproductive Biology, NYU School of Medicine, 180 Varick Street, New York, NY 10014, USA e Infertility Center of St Louis, St Luke’s Hospital, St Louis, MO 63017, USA f Reprogenetics, 3 Regent Street, Livingston, NJ 07078, USA g Department of Obstetrics and Gynecology, The Mount Sinai Hospital, E 101st Street, New York, NY 10029, USA

Dr John Zhang completed his medical degree at Zhejiang University School of Medicine in China, and subse-

quently received his Master’s Degree at Birmingham University in the UK. In 1991, Dr Zhang earned his PhD in

IVF, and, after researching the biology of mammalian reproduction and human embryology for nearly 10 years

he completed his fellowship training in Reproductive Endocrinology and Infertility at New York University’s School

of Medicine in 2001. Dr. Zhang continues his clinical research in minimal stimulation IVF, non-embryonic stem

cell, long-term oocyte cryopreservation, and oocyte reconstruction by nuclear transfer.

KEY MESSAGE

We report a live birth after oocyte spindle transfer to prevent transmission of the mitochondrial disease, Leigh

syndrome.

A B S T R A C T

Mutations in mitochondrial DNA (mtDNA) are maternally inherited and can cause fatal or debilitating mitochondrial disorders. The severity of clinical symptoms is often

associated with the level of mtDNA mutation load or degree of heteroplasmy. Current clinical options to prevent transmission of mtDNA mutations to offspring are limited.

Experimental spindle transfer in metaphase II oocytes, also called mitochondrial replacement therapy, isa novel technology for preventing mtDNA transmission from oocytes to

pre-implantation embryos. Here, we report a female carrier of Leigh syndrome (mtDNA mutation 8993T > G), with a long history of multiple undiagnosed pregnancy losses and

deaths of offspring as a result of this disease, who underwent IVF after reconstitution of her oocytes by spindle transfer into the cytoplasm of enucleated donor oocytes. A male

euploid blastocyst was

H.R.2029 Consolidated Appropriations Act, 2016

114th Congress

• (Sec. 749) Prohibits the FDA from acknowledging applications for an exemption for investigational use of a drug or biological product in research in which a human embryo is intentionally created or modified to include a heritable genetic modification. Provides that any submission is deemed not to have been received, and the exemption may not go into effect.

Do We Have a Moral Obligation to

Genetically Enhance our Children?

https://www.thehastingscenter.org/moral-obligation-genetically-enhance-children/

By Ronald M. Green

The Oxford philosopher Julian Savulescu, among others, has argued that prospective parents engaging in embryo selection using preimplantation genetic diagnosis not only may seek to have

genetically enhanced children but are morally obligated do so. (See, for example, his essay “Procreative Beneficence: Why We Should Select the Best Children,” Bioethics, 15, no.5/6, 2001.)

I argue that Savulescu is wrong.

Savulescu defends a moral principle he calls the principle of procreative beneficence. He states that, under this principle, prospective parents choosing among embryos “should select the child, of the

possible children they could have, who is expected to have the best life, or at least as good a life as the others, based on the relevant, available information.” Among the possible enhancements he

identifies are intelligence, memory, self-discipline, impulse control, foresight, patience, and a sense of humor. Savulescu has not yet extended this principle beyond preimplantation genetic diagnosis

to gene editing. He acknowledges that gene editing currently carries health risks not associated with embryo selection, and that these risks outweigh any obligation to try to bring about “the best life”

for a child.

But given the speed with which CRISPR technology is advancing, it seems that we are not far away from safe, effective gene enhancements for some traits. So according to Savulescu’s principle, when

sufficient levels of safety are reached, all parents with the means to afford gene editing for enhancement will have a moral obligation to do so.

I will specify my criticisms of Savulescu’s principle, but first I want to say that I fully support the reproductive use of gene editing technology for the prevention and elimination of serious genetic diseases.

If we could use gene editing to remove the gene sequences in an embryo that cause sickle cell disease or cystic fibrosis, I would say not only that we may do so, but in the case of such severe diseases, that

we have a moral obligation to do so.

I think that parents and medical professionals should always try to give a child a healthy start in life. This principle underlies the very firm moral intuition that pregnant women should not take drugs or

drink alcohol to excess during pregnancy. In an era of safe gene editing, I believe it would extend to the obligation to use this technology to avoid transmitting grave inherited disease conditions.

5/17/2017 Babies From Skin Cells? Prospect Is Unsettling to Some Experts The New York Times

https://nyti.ms/2ro5EhG

https://www.nytimes.com/2017/05/16/health/ivgreproductive-technology.html?hpw&rref=health&action=click&pgtype=Homepage&module=wellregion&region=bottom-

well&WT.nav=bottomwell&_r=0

27/5

HEALTH

Babies From Skin Cells? Prospect Is Unsettling to Some Experts By TAMAR LEWIN MAY 16, 2017

Nearly 40 years after the world was jolted by the birth of the first testtube baby, a new revolution in reproductive

technology is on the horizon — and it promises to be far more controversial than in vitro fertilization ever was.

Within a decade or two, researchers say, scientists will likely be able to create a baby from human skin cells that

have been coaxed to grow into eggs and sperm and used to create embryos to implant in a womb.

The process, in vitro gametogenesis, or I.V.G., so far has been used only in mice. But stem cell biologists say it is

only a matter of time before it could be used in human reproduction — opening up mindboggling possibilities.

With I.V.G., two men could have a baby that was biologically related to both of them, by using skin cells from one

to make an egg that would be fertilized by sperm from the other. Women with fertility problems could have eggs

methylation site H3K9me3 of SCNT embryos was further

confirmed by immunostaining of H3K9me3 (Figure 2F), showing

that the level of H3K9me3 was high in control one-cell SCNT

embryos but greatly reduced in those injected with Kdm4d

mRNA.

To identify candidate genes that were repressed by H3K9me3

and may be responsible for the poor development of monkey

SCNT embryos, we further examined the RNA-seq data and

found that some developmental pluripotency-associated genes

such as Dppa2, Dppa4, and Myc were repressed by H3K9me3

in monkey SCNT embryos. Other repressed genes found in

mouse SCNT embryos, such as Zscan4 and Polr3h, were also

identified in monkey SCNT embryos. A more complete list of

genes identified is shown in Data S1. Further studies on directly

manipulating these genes in SCNT embryos are needed to

confirm their roles in facilitating embryonic development (Ma-

toba et al., 2014). Taken together, the expression of H3K9me3

demethylase Kdm4d significantly improved epigenetic reprog-

ramming in monkey SCNT embryos similar to that found in other

species (Chung et al., 2015; Matoba et al., 2014). Thus, Kdm4d

mRNA injection was used in all subsequent experiments in the

present study.

SCNT Using Fetal Monkey Fibroblasts

Fetal fibroblasts in primary culture derived from an aborted

female cynomolgus monkey fetus were prepared by standard

methods (see STAR Methods) and used for SCNT. These cells

were chosen for their potential in obtaining a large number of

nuclei with uniform genetic background. Using the SCNT pro-

tocol described above, a total of 109 Kdm4d mRNA-injected

SCNT embryos under I/D/T condition were obtained using

127 MII-stage oocytes, and 79 of them (between 2-cell to

blastocyst stage) were transferred to 21 cynomolgus female

surrogates (Sun et al., 2008) (Table 1 and Data S2). The choice

of the embryo transfer time was based on the number of SCNT

embryos prepared and that of surrogates available at the time.

Pregnancy was confirmed in 6 surrogates by ultra- sound

examination one month later, 4 of them carried 5 fe- tuses (one

twin), and the other 2 carried only gestational sacs (GS,

Figures 3C and 3D). Among the 4 pregnancies, 2 aborted at

the early gestation stage (within two months), and 2 developed

beyond 140 days (Figure 3E and Data S2). Two live babies

were obtained at full term (155 and 141 days) by caesarean

section. The newborn baby monkeys, named Zhong Zhong

(ZZ) and Hua Hua (HH) (Figures 4A and

Figure 2. Blastocyst Development of SCNT

Monkey Embryos

(A and C) Percentages of cleaved embryos that

developed into blastocysts (blastocyst rate) of

SCNT embryos using fetal fibroblasts (A) and adult

cumulus cells (C) under different conditions. I:

ionomycin, D: 6-dimethylaminopurine, T: TSA, K:

Kdm4d mRNA.

(B and D) Percentages of blastocysts showing

prominent ICM in SCNT embryos using fetal

fibroblasts (B) and adult cumulus cells (D).

(E)Upregulation of RRRs resulting from Kdm4d

modification in SCNT embryos. CC: Expression

levels of RRRs in native donor cumulus cells. ICSI:

Expression levels of RRRs in eight cell-stage

monkey embryos obtained by intracytoplasmic

sperm injection (ICSI), indicating normal develop-

mental upregulation of RRRs. SC: expression

levels of RRRs in cumulus cells derived eight cell-

stage control SCNT embryos in the absence of

Kdm4d mRNA injection, showing low-level

expression of RRRs. SK: expression levels of

RRRs in cumulus cells derived eight cell-stage

SCNT embryos injected with Kdm4d mRNA,

showing elevated expression of many RRRs, some

of which correspond to those of ICSI eight cell-

stage embryos.

(F)Representative nuclear images of cumulus cells

derived one-cell stage SCNT embryos stained with

anti-H3K9me3 and DAPI at 5 hr after Kdm4d

mRNA injection. Scale bar, 30 mm.

Table 1. Statistics on the Development of SCNT Embryos

Donor cells Oocytes SCNT embryos Embryos transferred Surrogates Pregnancies Live birth Survived offspring

Fetal fibroblasts 127 109 79 21 6 2 2

Cumulus cells 290 192 181 42 22 2 0

Cell 172, 1–7, February 8, 2018 3

Please cite this article in press as: Liu et al., Cloning of Macaque Monkeys by Somatic Cell Nuclear Transfer, Cell (2018), https://doi.org/

10.1016/j.cell.2018.01.020

OOCYTES NT EMB ETS SURRO PRG LIVE BIRTH SURVIVAL

FETAL 127 109 79 21 6 2 2

ADULT 290 192 181 42 22 2 0

Surviving birth/ fetal oocytes 2 of 127 1.57%

Surviving birth/ adult oocytes 0 of 290 0%

Surviving birth/ fetal per NT embryo 2 1.83%

Surviving birth/ fetal per embryo transf 2.53%

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