Chapter 16. Genetics. Will Obs

 Chapter 16. Genetics

BS. Nguyễn Hồng Anh

Genetics is the study o genes, heredity, and the variation o

inherited characteristics. Medical genetics addresses the etiology and pathogenesis o human diseases that are at least

partially genetic in origin, as well as their prediction and prevention. Whereas a gene is a specic sequence o deoxyribonucleic acid (DNA) on a single chromosome that codes or

a particular protein, a genome is the entirety o all genes that

make up an organism. Genomics is the study o how genes

unction and interact with one other. Chromosomal, mendelian, and nonmendelian genetic conditions are reviewed in

this chapter. Prenatal and preimplantation genetic testing and

newborn genetic screening are discussed in Chapters 17 and

32, respectively.

Genetic disease is common. wo to 3 percent o newborns

have a recognized structural abnormality, another 3 percent are

diagnosed with an abnormality by age 5 years, and yet another

8 to 10 percent are ound to have a unctional or developmental abnormality beore reaching adulthood.

GENOMICS IN OBSTETRICS

Te Human Genome Project was an international research

program that sequenced the 3 billion base pairs and more

than 20,000 genes that make up the human genome. It paved

the way or research into gene organization and unction in

an eort to understand the molecular basis o disease. More

than 99 percent o our DNA is identical. Te coding regions o

DNA—exons—constitute only 1.5 percent o the genome. An

exome is the entirety o all the exons that an organism contains.

Introns are DNA sequences involved in coding regulation and

make up 24 percent o the genome. Intergenic DNA composes

the remainder.

Our genetic code varies once every 200 to 500 base pairs,

usually as a single-nucleotide polymorphism (SNP). Tus,

whole genome sequencing and whole exome sequencing,

described later (p. 327), hold tremendous potential to urther

elucidate genetic variants in disease.

oward this goal, genetic and genomic databases are maintained by the National Center or Biotechnology Inormation

(2021). Tese are reely accessible and can be indispensable

to providers who oer counseling and testing or genetic

conditions. Te GeneReviews database has in-depth clinical

inormation or more than 800 genetic conditions, including diagnostic criteria and management considerations. Te

Genetic Testing Registry (GTR) database contains inormation

or nearly 80,000 genetic tests and instructions or specimen

collection and transport to individual laboratories throughout

the world (National Center or Biotechnology Inormation,

2021). Te National Library o Medicine (2021) has also

established a genetic inormation database, the MedlinePlus

Genetics, which is intended or the lay population. It provides

explanations o more than 1300 genetic conditions, a glossary

o genetic concepts, and links to more comprehensive medical

resources.

CHROMOSOMAL ABNORMALITIES

Chromosomal abnormalities gure prominently in genetic disease. Tey account or >50 percent o rst-trimester miscarriages, approximately 20 percent o second-trimester losses, and

6 to 8 percent o stillbirths and early-childhood deaths (Reddy,

2012; Stevenson, 2004; Wou, 2016). Based on data rom

population-based registries, chromosomal abnormalities are

identied in 0.4 percent o recognized pregnancies (Wellesley,

2012). Most o these are aneuploidies. risomy 21 accounts or

>50 percent, trisomy 18 or nearly 15 percent, and trisomy 13

or 5 percent o cases (Fig. 16-1). I no karyotypic abnormality is identied in a etus with a structural anomaly, additional

testing with chromosomal microarray analysis is anticipated to

detect a chromosomal deletion or duplication—a copy number

variant—in approximately 6.5 percent o cases (American College o Obstetricians and Gynecologists, 2020c).

■ Standard Nomenclature

Chromosomal abnormalities are described using the International System or Human Cytogenomic Nomenclature

(McGowan-Jordan, 2020). Abnormalities all into two broad

categories: abnormal chromosome number, such as trisomy, and

abnormal chromosome structure, such as a DNA segment deletion or duplication. Each chromosome has a short arm, termed

the “p” or petit arm, and a long arm, known as the “q” arm,

selected because it is the next letter in the alphabet. Te two

arms are separated by the centromere.

When reporting a karyotype, the total number o chromosomes is listed rst, corresponding to the number o centromeres. Tis is ollowed by the sex chromosomes, XX or XY, and

then by a description o any structural variation. Specic chromosomal abnormalities are indicated by standard abbreviations,

such as del (deletion), inv (inversion), and t (translocation). Te

aected region or bands o the arms are then reported, so that

the reader will know the exact location. Examples are shown

in Table 16-1.

Nomenclature is similar or uorescence in situ hybridization (FISH), a technique used to rapidly identiy a specic

chromosome abnormality (p. 325). Te abbreviation ish applies

when in situ hybridization is perormed on metaphase cells and

nuc ish when perormed on interphase nuclei. I the result is

normal, ish is ollowed by the probe's specic chromosomal

region, such as 22q11.2, and then the name o the probe and

the number o signals visualized—or example, HIRAx2. I a

deletion is identied, del is included beore the chromosomal

region, and the name o the probe is ollowed by a minus sign

(HIRA–), as shown in able 16-1. Te 22q11.2 deletion syndrome is discussed later (p. 315).

For chromosomal microarray analysis (CMA), the designation begins with the abbreviation arr (p. 348). Tis is ollowed

by the version o the genome build to which the nucleotide

designations are aligned, such as GRCh38 or Genome Reerence Consortium human build 38. Next, the number o the

chromosome containing the abnormality is listed, ollowed by

the p or q arm, and then the specic bands in question. CMA

reports also include the aected base pair coordinates and thus

convey the exact size and location or every abnormality identi-

ed (see able 16-1). Te inormation is reported in the same

way whether the alteration is pathogenic or is o uncertain clinical signicance.

■ Abnormalities of Chromosome Number

Te most easily recognized chromosomal abnormalities are

numerical. Aneuploidy is the inheritance o either an extra

chromosome—resulting in trisomy, or loss o a chromosome—

monosomy. Tis diers rom polyploidy, which is an abnormal

number o whole haploid chromosome sets. For example, triploidy has 69 chromosomes. Te estimated incidence o various

numerical chromosomal abnormalities is shown in Figure 16-1

(Wellesley, 2012).

Autosomal Trisomies

risomy usually results rom nondisjunction, which is the

ailure o normal chromosomal pairing and separation during

meiosis. Nondisjunction may occur i the chromosomes: (1)

ail to pair up, (2) pair up properly but separate prematurely, or

(3) ail to separate. Although each chromosome pair is equally

likely to have a segregation error, trisomies other than 21, 18,

or 13 rarely result in term pregnancies. Each o these autosomal

54%

17%

5%

8%

5%

13%

Trisomy 21 (23:10,000)

Other (7:10,000)

47,XXX; 47,XXY; 47,XYY

(2:10,000)

45,X (3:10,000)

Trisomy 13 (2:10,000)

Trisomy 18 (6:10,000)

FIGURE 16-1 Prevalence and relative proportion of selected chromosomal abnormalities from EUROCAT (European Surveillance of

Congenital Anomalies) population-based registries that included

>10,000 aneuploid live births, fetal deaths, and pregnancy terminations, 2000–2006.310 The Fetal Patient

Section 6

TABLE 16-1. Examples of Karyotype Designations Using the 2020 International System for Human Cytogenomic

Nomenclature

Karyotype Description

46,XX Normal female chromosome constitution

47,XY,+21 Male with trisomy 21

47,XX,+21/46,XX Female who is a mosaic of trisomy 21 cells and cells with normal constitution

46,XY,del(4)(p14) Male with terminal deletion (del) of the short arm of chromosome 4 at band p14

46,XX,dup(5)(p14p15.3) Female with duplication (dup) of the short arm of chromosome 5 from band p14 to

band p15.3

45,XY,der(13;14)(q10;q10) Male with balanced robertsonian translocation (der) of the long arms of chromosomes

13 and 14—the karyotype now has one normal 13, one normal 14, and the

translocation chromosome, reducing the normal 46 chromosome complement to 45

46,XX,t(11;22)(q23;q11.2) Female with a balanced reciprocal translocation (t) between chromosomes 11 and 22,

with breakpoints at 11q23 and 22q11.2

46,XY,inv(3)(p21q13) Male with inversion (inv) of chromosome 3 that extends from p21 to q13—a pericentric

inversion because it includes the centromere

46,X,r(X)(p22.1q27) Female with one normal X and one ring (r) X chromosome, with the regions distal to

p22.1 and q27 deleted from the ring

46,X,i(X)(q10)

ish 22q11.2(HIRAx2)

ish del(22)(q11.2q11.2) (HIRA-)

arr[GRCh38] 18p11.32q23

(102328_79093443)x3

arr[GRCh38] 4q32.2q35.1

(163146681_183022312)x1

arr[GRCh38] 15q11.2q26

(23123715_101888908)x2 hmz

Female with one normal X chromosome and an isochromosome (i) of the long arm of

the other X

FISH of metaphase cells using a probe for the HIRA locus of the 22q11.2 region, with 2

signals identified (no evidence of microdeletion)

FISH of metaphase cells using a probe for the HIRA locus of the 22q11.2 region, with

only one signal identified, consistent with the microdeletion

Microarray analysis (arr), genome build GRCh38, showing a single copy gain on

chromosome 18 from band p11.32 to band q23 (essentially the entire chromosome),

consistent with trisomy 18

Microarray analysis (arr), genome build GRCh38, showing a copy loss on the long arm of

chromosome 4 at bands q32.2 through q35.1 (19.9 Mb)

SNP microarray analysis (arr), genome build GRCh 38, showing homozygosity for the

entire long arm of chromosome 15

FISH = fluorescence in situ hybridization; GRCh38 = Genome Reference Consortium human build 38; HIRA = histone cell

cycle regulator; SNP = single nucleotide polymorphism.

Used with permission from Dr. Kathleen S. Wilson.

trisomies has a spectrum o phenotypic severity. Some etuses

come to attention earlier in a pregnancy due to multiple major

organ abnormalities or hydrops etalis (p. 328). Others have

more subtle ndings or are detected only through prenatal

screening tests (Chap. 17, p. 337). Survival data are based on

cases with better prognosis, and this actors into counseling.

Te likelihood that a pregnancy will be complicated by an

autosomal trisomy rises steeply with maternal age and particularly ater age 35 (Fig. 16-2) (Mai, 2013). From birth until

ovulation, oocytes are suspended in midprophase o meiosis I. I

nondisjunction occurs ater completion o meiosis, one gamete

will have two copies o the aected chromosome, which leads

to trisomy i ertilized. Te other gamete, receiving no copy o

the aected chromosome, is nullisomic and will be monosomic

i ertilized. It is estimated that 10 to 20 percent o oocytes

are aneuploid secondary to meiotic errors compared with 3

to 4 percent o sperm. Ater a pregnancy with an autosomal

trisomy, the risk or any autosomal trisomy in a uture pregnancy approximates 1 percent until the woman's age-related

risk exceeds this. Screening and prenatal diagnosis o autosomal

0

<20

Prevalence per 10,000 live births

Maternal age at delivery (years)

20–24 25–29 30–34 35–39 40 or older

20

60

40

80

120

100

140

Trisomy 21

Trisomy 18

Trisomy 13

FIGURE 16-2 Prevalence of autosomal trisomies according to

maternal age found in population-based birth defect surveillance

programs in the United States, 2006–2010.

trisomies are discussed in Chapter 17 (p. 333). Parental chromosomal studies are not indicated unless the abnormality was

caused by an unbalanced translocation or other structural

re arrangement (p. 315).Genetics 311

CHAPTER 16

Trisomy 21—Down Syndrome. In 1866, J. L. H. Down

described a group o intellectually disabled children with distinctive physical eatures. Nearly 100 years later, Lejeune (1959)

demonstrated that Down syndrome stems rom an autosomal

trisomy (Fig. 16-3). risomy 21 causes 95 percent o Down

syndrome cases. Te nondisjunction event occurs during meiosis I in approximately 75 percent o cases and in meiosis II in

the remainder. Because chromosome 21 is acrocentric, a robertsonian translocation (p. 315) may occur, and this accounts

or 3 to 4 percent o Down syndrome births. Te remaining 1

to 2 percent are caused by isochromosome 21 or mosaicism or

chromosome 21 (p. 317).

Down syndrome is the most common nonlethal trisomy.

It is identied in approximately 1 in 450 pregnancies (Loane,

2013). In the United States, etal losses and pregnancy terminations yield an estimated prevalence o 1 case per 740 live births

(Mai, 2013; Parker, 2010). Te etal death rate beyond 20

weeks’ gestation approximates 5 percent. Coinciding with the

rise in overall number o pregnancies in women aged 35 years

and older, the proportion o pregnancies with Down syndrome

has increased approximately 33 percent during the past our

decades (Loane, 2013; Parker, 2010; Shin, 2009). Most Down

syndrome births at Parkland Hospital now occur in women in

this age group (Hussamy, 2019).

I major anomalies and minor aneuploidy markers are considered, 50 to 75 percent o pregnancies aected by Down syndrome are ound to have a sonographic abnormality (Chap. 17,

p. 340) (American College o Obstetricians and Gynecologists, 2020d; Hussamy, 2019). A major structural malormation is detected in the second trimester in approximately one

third o cases. Te most prevalent anomalies in aected children

are cardiac and gastrointestinal. Cardiac abnormalities occur

in approximately 50 percent, particularly endocardial cushion

deects and ventricular septal deects (Figs. 15-39A and 15-39B,

p. 291) (Bergstrom, 2016; Freeman, 2008; Stoll, 2015). Gastrointestinal abnormalities are identied in 6 to 12 percent and

include esophageal atresia, duodenal atresia, and Hirschsprung

disease (Fig. 15-48, p. 296) (Bull, 2011; Stoll, 2015).

Other health problems are also common with Down syndrome. Tese include hearing loss in 75 percent, optical reractive errors in 50 percent, cataracts in 15 percent, obstructive

sleep apnea in 60 percent, thyroid disease in 15 percent, transient myeloprolierative disorder in 10 percent o newborns,

early-onset Alzheimer's disease in 70 percent o adults, and

a higher incidence o leukemia (Bull, 2011; Hartley, 2015).

Women with Down syndrome are ertile, and nearly one third

o their ospring will have Down syndrome. Males with Down

syndrome have markedly reduced spermatogenesis and are

almost always sterile. Te average intelligence quotient (IQ)

score is 35 to 70. Social skills in aected children are usually

higher than predicted by their IQ scores.

Characteristic eatures o Down syndrome are shown in

Figure 16-4. ypical ndings include brachycephaly; epicanthal olds and up-slanting palpebral ssures; Brusheld spots,

which are whitish spots on the periphery o the iris; a at nasal

bridge; and hypotonia. Inants oten have loose skin at the

FIGURE 16-3 Male karyotype with trisomy 21 (47,XY,+21), consistent with Down syndrome. (Reproduced with permission from

Dr. Prasad Koduru.)

A B C

FIGURE 16-4 Newborn with Down syndrome, karyotype 47,XX,+21 (trisomy 21). A. Characteristic facial features include epicanthal folds

and a flattened nasal bridge. B. The hand has a single palmar crease (arrow) and hypoplasia of the middle phalanx of the fifth digit (arrowhead). C. The “sandal gap” between the first and second toes is also known as hallux varus. (Reproduced with permission from Dr. Aldeboran N. Rodriguez.)312 The Fetal Patient

Section 6

nape o the neck, a prominent space or “sandal-gap” between

the rst and second toes, short ngers, a single palmar crease,

and clinodactyly o the th digit, in which hypoplasia o the

middle phalanx causes inward curvature. Discussed in Chapter

17, some o these ndings are seen during prenatal ultrasound

examination.

Data suggest that approximately 95 percent o liveborn

inants with Down syndrome survive the rst year. Te 10-year

survival rate is at least 90 percent overall and is 99 percent

i major malormations are absent (Rankin, 2012; Vendola,

2010). Average lie expectancy is 55 to 60 years. Several organizations oer education and support or prospective parents aced with prenatal diagnosis o Down syndrome. Tese

include the March o Dimes, National Down Syndrome Congress (www.ndsccenter.org), and National Down Syndrome

Society (www.ndss.org).

Trisomy 18—Edwards Syndrome. Te association between

this constellation o abnormalities and an autosomal trisomy

was rst described by Edwards (1960). In population-based

series o abortuses, stillbirths, and live births, trisomy 18 approximates 1 per 2000 recognized pregnancies (Goel, 2019; Loane,

2013; Parker, 2010). High in-utero lethality and termination

o many aected pregnancies explains a live birth prevalence o

just 1 per 6600 to 10,000. Unlike Down syndrome and Patau

syndrome, which may result rom a robertsonian

translocation because they

involve acrocentric chromosomes, Edwards syndrome uncommonly stems

rom a chromosomal rearrangement.

Virtually every organ

can be aected by trisomy 18. Common major

anomalies include heart

deects in more than 90

percent, particularly ventricular septal deects.

Cerebellar vermian agenesis, myelomeningocele,

diaphragmatic hernia, omphalocele, imperorate anus,

and renal anomalies such

as horseshoe kidney are

others (Rosa, 2011; Springett, 2015; Watson,

2008). Sonographic images

o these abnormalities are

shown in Chapter 15.

Frequent cranial and extremity abnormalities in

aected etuses include a

“strawberry-shaped” cranium, abnormally wide

cavum septum pellucidum, choroid plexus cysts,

micrognathia, clenched hands with overlapping digits, radial

aplasia with hyperexed wrists, and rockerbottom or clubbed

eet (Fig. 16-5) (Abele, 2013). Importantly, choroid plexus

cysts only raise the risk or trisomy 18 in the setting o other

risk actors, such as etal structural abnormalities or an abnormal aneuploidy screening test result (Reddy, 2014).

Pregnancies with trisomy 18 that reach the third trimester oten develop etal-growth restriction, and the mean

birthweight is <2500 g (Lin, 2006; Rosa, 2011). When

undiagnosed, trisomy 18 has resulted in emergency cesarean

or “etal distress” in nearly 50 percent o cases (Schneider,

1981; Houlihan, 2013). Mode o delivery and management

o heart rate abnormalities should be discussed in advance.

Discussed in Chapter 35 (p. 628), perinatal palliative care

consultation should be oered (American College o Obstetricians and Gynecologists, 2019).

In the National Birth Deects Prevention Study, median

neonatal survival with trisomy 18 was 8 days, and the 5-year

survival rate was 12 percent (Meyer, 2016). Not unexpectedly,

major anomalies lower survival rates. Data rom the Society o

Toracic Surgeons indicate that inants with trisomy 18 who

undergo cardiac surgery have mortality rates 3 to 5 times higher

than those without trisomy 18 (Cooper, 2019). Tus, among

the minority with trisomy 18 who survive until birth, the condition is severely lie limiting.

A B

C D

FIGURE 16-5 Trisomy 18—Edwards Syndrome. A. This transventricular sonographic view shows fetal

choroid plexus cysts and an angulated “strawberry-shaped” skull. B. Radial clubhand is manifested as a

single forearm bone (radius), with the hands in a fixed, hyperflexed position at right angles to the forearms.

C. This three-dimensional (3-D) sonographic image shows the characteristic hand position of clenched fist

with overlapping digits. D. 3-D sonographic image displays a rockerbottom foot (vertical talus).Genetics 313

CHAPTER 16

Trisomy 13—Patau Syndrome. Tis constellation o etal abnormalities and their association with an autosomal trisomy was rst

described by Patau and colleagues (1960). Te incidence o trisomy 13 approximates 1 case in 5000 recognized pregnancies,

which includes abortuses and stillbirths. Most aected etuses are

lost or terminated. Te live birth prevalence is 1 per 12,000 to

18,000 (Goel, 2019; Loane, 2013; Parker, 2010).

At least 80 percent o pregnancies with Patau syndrome

result rom trisomy 13. With rare exception, the remainder are

caused by a robertsonian translocation involving chromosome

13. Te most common translocation involves chromosomes

13 and 14, der(13;14)(q10;q10), which is carried by approximately 1 in 1300 phenotypically normal individuals. Among

translocation carriers, ewer than 2 percent give birth to a live

inant with Patau syndrome.

risomy 13 may be associated with abnormalities o any

organ system. Abnormalities o the brain, heart, kidneys, and

extremities are the most requent. Holoprosencephaly, usually the alobar type, is present in two thirds o cases and oten

is accompanied by severe acial abnormalities (Fig. 15-13,

p. 279). Tese may include hypotelorism or cyclopia, microphthalmia, and nasal abnormalities that range rom a single

nostril to a proboscis. Cardiac deects are ound in up to 90

percent (Shipp, 2002). Other abnormalities that suggest trisomy 13 include cephalocele, microcephaly, omphalocele,

cystic renal dysplasia, polydactyly, rockerbottom eet, aplasia

cutis, and clet lip-palate, which may be median (Lin, 2007;

Springett, 2015). Sonographic images o several o these are

shown in Chapter 15. Aected etuses oten also have bilateral

echogenic intracardiac oci. For the etus or newborn with a

cephalocele, cystic kidneys, and polydactyly, the dierential diagnosis includes trisomy 13 and the autosomal-recessive Meckel–

Gruber syndrome.

risomy 13 is lie limiting, even more so than trisomy

18 (Domingo, 2019). In the National Birth Deects Prevention Study, median survival o neonates with trisomy 13 was

5 days, and the 5-year survival rate was below 10 percent

(Meyer, 2016). Occasionally children with trisomy 13 have

been candidates or palliative surgical procedures. Most o these

have a milder phenotype that lacks brain, cardiac, gastrointestinal, or genitourinary abnormalities (Nelson, 2016). Counseling

regarding prenatal diagnosis and management options is similar

to that described or trisomy 18.

Unlike other aneuploidies, etal trisomy 13 coners risk to

the pregnant woman. Continuing pregnancies have at least a

25-percent risk or hypertensive complications (Dotters-Katz,

2018; uohy, 1992). Te risk or preeclampsia with severe eatures is increased more than tenold and oten develops prior

to 32 weeks’ gestation. Chromosome 13 contains the gene or

soluble ms-like tyrosine kinase-1 (sFlt-1), which is an antiangiogenic protein associated with preeclampsia (Chap. 40,

p. 694). Investigators have documented overexpression o the

sFlt-1 protein by trisomic 13 placentas and in serum o women

with preeclampsia (Bdolah, 2006; Silasi, 2011).

Other Trisomies. Live births are extremely rare with any other

autosomal trisomy. risomy 22 has been described in case

reports and series (Heinrich, 2013; Kehinde, 2014). Aected

etuses have severe growth restriction, microcephaly with an

abnormal-shaped cranium, midace hypoplasia, and cardiac

and genitourinary abnormalities. risomy 16 is the most prevalent trisomy among rst-trimester losses, accounting or 16

percent. risomies 21 and 22 are also common among rst

trimester losses. risomy 1 has never been reported.

Monosomy

Nondisjunction creates an equal number o nullisomic and

disomic gametes. As a rule, missing chromosomal material is

more devastating than extra chromosomal material, and almost

all monosomic conceptuses are lost beore implantation. Te

one exception is monosomy or the X chromosome (45,X),

urner syndrome, which is discussed subsequently. Unlike

autosomal trisomies, which increase in incidence with maternal

age, monosomy and maternal age lack an association.

Polyploidy

Tis is dened as more than two complete haploid chromosomal sets. Polyploidy accounts or approximately 20 percent

o spontaneous abortions but is rarely ound later in gestation.

riploid pregnancies have three haploid sets, 69 chromosomes. I extra set o chromosomes is paternal, the result is

diandric triploidy, and i maternal, digynic triploidy, relevant

because the phenotype reects the parent o origin. Tis is an

example o imprinting (p. 322). Diandric or type I triploidy

occurs ater ertilization o one egg by two sperm or by one

abnormal diploid sperm. Diandric triploidy produces a partial

molar pregnancy, discussed in Chapter 13 (p. 237). With digynic or type II triploidy, the extra chromosomal set is maternal,

and the egg ails to undergo the rst or second meiotic division

beore ertilization. Digynic triploid placentas do not develop

molar changes. However, the etus usually develops asymmetrical growth restriction. Although diandric triploidy accounts or

the majority o triploid conceptions, the early loss rate is so

high that two thirds o triploid pregnancies identied beyond

the rst trimester are digynic ( Jauniaux, 1999).

riploidy is recognized in 1 per 5000 pregnancies (Zalel,

2016). It is considered lethal, and etuses with either the diandric or digynic orm typically have multiple structural anomalies. Te brain, heart, kidneys, and extremities are commonly

aected (Massalska, 2017; Zalel, 2016). Counseling, prenatal

diagnosis, and delivery management are similar to those or

trisomies 18 and 13. Te recurrence risk or a woman whose

triploid etus survived past the rst trimester is 1 to 1.5 percent,

and thus prenatal diagnosis is oered in uture pregnancies.

etraploid pregnancies have our haploid sets o chromosomes,

resulting in either 92,XXXX or 92,XXYY. Tis suggests a postzygotic ailure to complete an early cleavage division. Te conceptus

invariably succumbs, and the recurrence risk is minimal.

Sex Chromosome Abnormalities

45,X—Turner Syndrome. First described by urner (1938),

this syndrome later was ound to be caused by monosomy

X (Ford, 1959). Te birth prevalence o urner syndrome is

approximately 1 in 2500 girls (Cragan, 2009; Dolk, 2010). Te

missing X chromosome is paternally derived in 80 percent o

cases (Cockwell, 1991; Hassold, 1990).314 The Fetal Patient

Section 6

urner syndrome is the only monosomy compatible with

lie, but it is also the most common aneuploidy in rst-trimester

losses, accounting or 20 percent. Tis is explained by its wide

range in phenotype. Approximately 98 percent o aected conceptuses are so abnormal that they abort early in the rst trimester. O the remainder, many maniest large, septated cystic

hygromas in the late rst or early second trimester, oten in

the setting o edema that progresses to hydrops etalis. When

cystic hygromas are accompanied by hydrops, the prognosis is

extremely poor (Chap. 15, p. 286).

Fewer than 1 percent o pregnancies with urner syndrome

result in a liveborn neonate. O these, only hal are actually

monosomy X—approximately a ourth have mosaicism or

monosomy X, such as 45,X/46,XX or 45,X/46,XY, and another

15 percent have isochromosome X. It is thought that surviving

individuals with 45,X may have had the benet o “rescue” by

an additional cell line containing 46,XX during critical phases

o development that was subsequently lost (Hook, 2014).

Abnormalities associated with urner syndrome include

let-sided cardiac deects—such as coarctation o the aorta,

hypoplastic let heart syndrome, or bicuspid aortic valve—in 30

to 50 percent; renal anomalies, particularly horseshoe kidney;

and hypothyroidism. Other eatures are short stature, broad

chest with widely spaced nipples, congenital lymphedema—

pufness over the dorsum o hands and eet, and a “webbed”

posterior neck resulting rom cystic hygromas. Intelligence

scores are generally in the normal range, but there may be

impairments in visual-spatial organization, nonverbal problem

solving, and interpretation o social cues. A consensus guideline is available that addresses screening and treatment or the

range o health problems aected individuals ace (Graveholt,

2017). Growth hormone is typically administered in childhood

to ameliorate short stature. More than 90 percent have ovarian dysgenesis and require estrogen repletion at puberty. An

exception is mosaicism involving the Y chromosome, as this

coners risk or germ cell neoplasm—regardless o whether the

child is phenotypically male or emale. Accordingly, eventual

prophylactic bilateral gonadectomy is indicated (Cools, 2011;

Schorge, 2020).

47,XXX. Approximately 1 in 1000 emale newborns has an

additional X chromosome—47,XXX (Berglund, 2020). Te

extra X is maternally derived in more than 90 percent o cases.

Te prevalence o 47,XXX is weakly associated with maternal

age, and cell-ree DNA screening has resulted in increased diagnoses. No specic pattern o malormations has been described,

but genitourinary problems and seizure disorders are more

common (Wigby, 2016). Aected inants do not have a characteristic appearance. When children do come to attention, eatures may include tall stature, hypertelorism, epicanthal olds,

kyphoscoliosis, clinodactyly, and hypotonia (artaglia, 2010;

Wigby, 2016). More than one third are diagnosed with a learning disability, hal have attention decit disorder, and overall

cognitive scores are in the low-average range. Pubertal development is unaected. Primary ovarian insufciency has been

reported. In the absence o prenatal diagnosis, it is estimated

that 47,XXX is ascertained in only 10 percent o aected children (artalgia, 2010).

Females with two or more extra X chromosomes—48,XXXX

or 49,XXXXX—are likely to have physical abnormalities apparent at birth. Tese abnormal X complements are associated

with intellectual disability. For both males and emales, the IQ

score is lower with each additional X chromosome.

47,XXY—Klinefelter Syndrome. Tis is the most common

sex chromosome abnormality. It occurs in approximately 1 in

700 male inants (Radicioni, 2010). Te additional X chromosome is maternally or paternally derived with equal propensity

(Jacobs, 1995; Lowe, 2001). Tere is a weak association with

advanced maternal and paternal age.

Like 47,XXX, newborns with 47,XXY usually appear phenotypically normal and do not have a higher incidence o anomalies. As children, boys are typically taller than average and have

normal prepubertal development. However, they have gonadal

dysgenesis, do not undergo normal virilization, and require testosterone supplementation. Tey may develop gynecomastia.

IQ scores usually lie in the average to low-average range, with

increased rates o delayed language development (Boada, 2009;

Girardin, 2011). Initiation o hormone replacement was previously recommended to begin in adolescence. However, more

recent research suggests that therapy earlier in childhood results

in improved working memory and executive unctioning and

a decrease in anxiety disorders (Samango-Sprouse 2019; ran,

2019).

47,XYY. Tis aneuploidy occurs in approximately 1 in 1000

male newborns (Berglund, 2020). As with 47,XXX and XXY

individuals, aected boys tend to be tall. A third have macrocephaly, nearly two thirds demonstrate hypotonia, and tremors are common (Bardsley, 2013). Rates o major anomalies

are not elevated, although hypertelorism and clinodactyly may

be identied in more than hal. Pubertal development is normal, and ertility is unimpaired. Aected individuals do have

increased rates o oral and written language impairments, attention decit disorder, developmental delays, and autism spectrum disorder (Bardsley, 2013; Joseph, 2018).

Males with more than two Y chromosomes—48,XYYY—

or with both additional X and Y chromosomes—48,XXYY or

49,XXXYY—are more likely to have congenital abnormalities,

medical problems, and intellectual disability (artaglia, 2011).

■ Abnormalities of Chromosome Structure

Structural chromosomal abnormalities include deletions, duplications, translocations, isochromosomes, inversions, ring chromosomes, and mosaicism (see able 16-1). Identication o

a structural chromosomal abnormality in ospring raises two

primary questions. First, what phenotypic or later developmental abnormalities are associated with the nding? Second,

is parental karyotype evaluation indicated? In other words, are

the parents at increased risk o carrying this abnormality, and i

so, what is their risk o having uture aected ospring?

Deletions and Duplications

A chromosome with a deletion has a stretch o DNA that is

missing, whereas one with a duplication has a region that is

included twice. Most deletions and duplications occur duringGenetics 315

CHAPTER 16

meiosis and result rom malalignment or mismatching during

the pairing o homologous chromosomes. Te misaligned segment may then be deleted, or i the mismatch remains when

the two chromosomes recombine, it may result in a deletion

in one chromosome and duplication in the other (Fig. 16-6).

When a deletion or duplication is identied in a etus or inant,

parental karyotyping should be oered, because i either parent carries a balanced translocation, the recurrence risk in

subsequent pregnancies is signicantly increased. Deletions

involving DNA segments large enough to be seen with standard cytogenetic karyotyping are identied in approximately

1 in 7000 births. Common deletions may be reerred to by

eponyms—or example, del 5p is called cri du chat syndrome.

Microdeletions and Microduplications. When a deletion or

duplication is smaller than 3 to 5 million base pairs, it is too

small to be detected with a standard karyotype analysis. CMA

permits identication o these microdeletions and microduplications (p. 325). When CMA is used, the region o DNA that is

missing or duplicated is termed a genomic copy number variant.

A microdeletion or duplication may involve a stretch o DNA

that contains multiple genes. Tis causes a contiguous gene syndrome, which can encompass serious but unrelated phenotypic

abnormalities (Schmickel, 1986). In some cases, a microduplication may involve the exact DNA region that causes a recognized microdeletion syndrome (Table 16-2). When a specic

microdeletion syndrome is suspected clinically, it is conrmed

using either CMA or FISH.

22q11.2 Microdeletion Syndrome. Tis syndrome is also

known as DiGeorge syndrome, Shprintzen syndrome, and

velocardioacial syndrome. It is the most common microdeletion, with a prevalence o 1 case in 3000 to 6000 births. Inheritance is autosomal dominant—ospring o aected individuals

have a 50-percent chance o inheriting the syndrome. However,

more than 90 percent o cases arise rom a de-novo mutation.

Te ull deletion contains 3 million base pairs, encompasses 40

genes, and includes 180 dierent eatures (Shprintzen, 2008).

Tis is emblematic o a contiguous gene syndrome and poses

counseling challenges because eatures can vary widely, even

among amily members. With prenatal diagnosis o an aected

etus, genetic testing is oered to the pregnant patient and her

partner.

Approximately 75 percent o aected individuals have an associated conotruncal cardiac anomaly, such as tetralogy o Fallot,

truncus arteriosus, interrupted aortic arch, or ventricular septal

deect (McDonald-McGinn, 2015). Immune deciency, such as

-cell lymphopenia, also develops in 75 percent. More than 70

percent have velopharyngeal insufciency or clet palate. Characteristic acial eatures include short palpebral ssures, bulbous

nasal tip, micrognathia, short philtrum, and small or posteriorly

rotated ears. Learning disabilities, autism spectrum disorder, and

intellectual disability also are common. Other maniestations

include hypocalcemia, renal anomalies, esophageal dysmotility,

hearing loss, behavioral disorders, and psychiatric illness, particularly schizophrenia.

Chromosomal Translocations

Tese are DNA rearrangements in which a DNA segment

breaks away rom one chromosome and attaches to another.

Te rearranged chromosomes are called derivative (der) chromosomes. Te two types are reciprocal and robertsonian translocations.

Reciprocal Translocations. A double-segment or reciprocal translocation results when breaks occur in two dierent

chromosomes and the broken ragments are exchanged. Each

aected chromosome receives a ragment o the other. I no

chromosomal material is gained or lost, the translocation is

considered balanced. Te prevalence o reciprocal translocations approximates 1 in 600 births.

Te carrier o a balanced translocation will usually have a

normal phenotype. However, repositioning o specic genes

within chromosomal segments may cause major structural or

developmental abnormalities in approximately 6 percent o

apparently balanced translocation carriers. Using CMA technology, up to 20 percent o individuals who appear to have

a balanced translocation are ound instead to have missing or

redundant DNA segments (Manning, 2010).

Balanced translocation carriers are at risk to produce unbalanced gametes, resulting in abnormal ospring. As shown in

Figure 16-7, i an oocyte or sperm contains a translocated chromosome, ertilization results in an unbalanced translocation—

monosomy or part o one aected chromosome and trisomy

or part o the other. In general, translocation carriers identied

ater the birth o an abnormal child have a 5- to 30-percent risk

o producing liveborn ospring with an unbalanced translocation. Carriers identied or other reasons, or example, during

an inertility evaluation, have only a 5-percent risk. Tis risk

is lower because gametes are so abnormal that conceptions are

nonviable.

Robertsonian Translocations. Tese involve only the acrocentric chromosomes—13, 14, 15, 21, and 22. Acrocentric

Meiosis I Meiosis II

Normal Del Dupl Normal

FIGURE 16-6 A mismatch during pairing of homologous chromosomes may lead to a deletion in one chromosome and a duplication in the other. Del = deletion; Dupl = duplication.316 The Fetal Patient

Section 6

chromosomes have extremely short p arms, and the p arms

contain redundant copies o genes coding or ribosomal RNA.

In a robertsonian translocation, the q arms o two acrocentric

chromosomes use at one centromere to orm a derivative chromosome, and the other centromere and both p arms are lost.

Te lost DNA is present in multiple copies on other acrocentric chromosomes, and thus the translocation carrier is usually

phenotypically normal. Because the number o centromeres

determines the chromosome count, a robertsonian translocation carrier has only 45 chromosomes.

Balanced robertsonian carriers have reproductive difculties.

During ertilization, i the derivative chromosome pairs with a

normal haploid chromosome, the resulting ospring will have

trisomy. I the used chromosomes are homologous, that is,

rom the same chromosome pair, the carrier can produce only

unbalanced gametes. Each egg or sperm contains either both

copies o the translocated chromosome, which would result in

trisomy i ertilized, or no copy, which would result in monosomy. I the used chromosomes are nonhomologous, our o

the six possible gametes would be abnormal.

Robertsonian translocations are ound in 1 in 1000 individuals. Te risk to have an abnormal ospring approximates

15 percent i a robertsonian translocation is carried by the

mother and 2 percent i carried by the ather. However, robertsonian translocations are not a major cause o miscarriage and

are identied in ewer than 5 percent o couples evaluated or

recurrent pregnancy loss. I a etus or child is ound to have a

translocation trisomy, both parents should be oered karyotype analysis. I neither parent is a carrier, the recurrence risk is

extremely low. Te most common robertsonian translocation

TABLE 16-2. Selected Microdeletion Syndromes

Syndrome Prevalence Location Features

Alagille 1:70,000 20p12.2 Cholestasis (paucity of intrahepatic bile ducts),

cardiac disease, skeletal disease, ocular

abnormalities, dysmorphic facies

Angelman 1:12,000 to 1:20,000 15q11.2–q13

(maternal genes)

Dysmorphic facies—“happy puppet” appearance,

intellectual disability, ataxia, hypotonia, seizures

Cri-du-chat 1:20,000 to 1:50,000 5p15.2–15.3 Abnormal laryngeal development with “cat-like”

cry, hypotonia, intellectual disability

Kallmann syndrome 1:30,000 males Xp22.3 Hypogonadotropic hypogonadism, anosmia

Langer-Giedion Rare 8q23.3 Trichorhinophalangeal syndrome, dysmorphic

facies, skeletal abnormalities, sparse hair

Miller-Dieker Rare 17p13.3 Neuronal migration abnormalities with

lissencephaly and microcephaly (profound

impairment), dysmorphic facies

Prader-Willi 1:10,000 to 1:30,000 15q11.2–q13

(paternal genes)

Obesity, hypotonia, hypogonadotropic

hypogonadism, small hands and feet,

intellectual disability

Retinoblastoma 1:280,000 13q14.2 Retinoblastoma, retinoma (benign neoplasm),

non-retinal (second primary) tumors

Rubenstein-Taybi 1:100,000 to 1:125,000 16p13.3 Dysmorphic facies, broad thumbs and toes,

intellectual disability, increased tumor risk

Smith-Magenis 1:15,000 to 1:25,000 17p11.2 Dysmorphic facies, speech delay, hearing loss,

sleep disturbances, self-destructive behaviors,

intellectual disability

Velocardiofacial

syndrome

1:3000 to 1:6000 22q11.2 Conotruncal cardiac defects, cleft palate,

velopharyngeal incompetence, thymic and

parathyroid abnormalities, intellectual disability

WAGR 1:500,000 11p13 Wilms tumor, aniridia, genitourinary anomalies.

intellectual disability (mental retardation)

Williams-Beuren 1:7500 to 1:10,000 7q11.23 Dysmorphic facies, dental malformation, aortic and

peripheral pulmonary artery stenosis, intellectual

disability

Wolf-Hirschhorn 1:20,000 to 1:50,000 4p16.3 Dysmorphic (“Greek helmet”) facies, delayed

growth and development, cleft lip/palate,

coloboma, cardiac septal defects

X-linked ichthyosis 1:6000 Xp22.3 Steroid sulfatase deficiency, corneal opacities

Prevalence reflects live births.

Data from National Library of Medicine, 2021; Johns Hopkins University, 2021.Genetics 317

CHAPTER 16

is der(13;14)(q10;q10), which accounts or up to 20 percent o

cases o Patau syndrome (p. 313).

Isochromosomes

When either two p arms or two q arms rom the same chromosome use together, an isochromosome results. Tis may occur

i the centromere breaks transversely instead o longitudinally

during meiosis II or mitosis. Alternately, an isochromosome can

result rom a meiotic error in a chromosome with a robertsonian

translocation. I the isochromosome is acrocentric, it is composed

o two q arms and will behave like a homologous robertsonian

translocation. Such a carrier is phenotypically normal but can

produce only abnormal, unbalanced gametes. I the isochromosome is nonacrocentric, it may be composed o either two p arms

or two q arms. Functionally this results in trisomy o the genes on

the arms that are present and monosomy o the genes on the arms

that are lost. Tus, the carrier is usually phenotypically abnormal

and produces abnormal gametes. Te most common isochromosome involves the long arm o the X chromosome, i(Xq), which is

the etiology o 15 percent o cases o urner syndrome.

Chromosomal Inversions

When the same chromosome breaks in two places, the intervening genetic material may invert beore the breaks are repaired.

Although no genetic material is lost or duplicated, an inversion

may alter gene unction. Tere are two types—pericentric and

paracentric inversions.

Pericentric Inversion. Tis results rom breaks in both the p and

q arms o a chromosome, such that the inverted material spans

the centromere (Fig. 16-8). A pericentric inversion causes problems in chromosomal alignment during meiosis and coners signicant risk or the carrier to produce abnormal gametes and

abnormal ospring. Te observed risk o abnormal ospring is

5 to 10 percent i ascertainment is made ater the birth o an

abnormal child. Te risk is only 1 to 3 percent i prompted by

another indication. An important exception is inv(9)(p11q12), a

pericentric inversion on chromosome 9 that is a normal variant

and present in approximately 1 percent o the population.

Paracentric Inversion. I one arm o a chromosome suers the

two breaks, the inverted material does not include the centromere, and the inversion is paracentric (see Fig. 16-8). Te carrier makes either normal balanced gametes or gametes that are

so abnormal as to preclude ertilization. Tus, although inertility may be a problem, the risk o having an abnormal ospring

is extremely low.

Ring Chromosome

I a deletion occurs at each end o the same chromosome, the

ends may come together to orm a ring chromosome. Te telomere regions at the ends o each chromosome contain redundant nucleoprotein complexes that stabilize the chromosome.

I only the telomeres are lost, all necessary genetic material is

retained, and the carrier is balanced. I a deletion extends more

Balanced translocation carrier

B Balanced

translocation

carrier

C Unbalanced

duplicationdeletion

D Unbalanced

duplicationdeletion

Normal

A Normal

Zygotes Gametes Parents

FIGURE 16-7 A carrier of a balanced translocation may produce offspring who are also carriers of the balanced rearrangement (B), offspring with unbalanced translocations (C, D), or offspring with normal chromosomal complements (A).318 The Fetal Patient

Section 6

should be oered. In a series o more than 1000 pregnancies with mosaicism at chorionic villus sampling, subsequent

amniocentesis identied true etal mosaicism in 13 percent.

Conned placental mosaicism accounted or 85 percent o

cases, and uniparental disomy, discussed later (p. 322), was

ound in 2 percent (Malvestiti, 2015). I mosaicism is detected

or a chromosome known to contain imprinted genes—such

as chromosomes 6, 7, 11, 14, or 15—testing or uniparental

disomy should be considered, as there may be etal consequences (Grati, 2014a).

Although outcomes with conned placental mosaicism are

generally good, risks o etal-growth restriction and stillbirth are

higher (Reddy, 2009). Fetal-growth restriction may stem rom

impaired unctioning o the aneuploid placental cells (Baero,

2012). In a recent series o 5500 pregnancies undergoing chorionic villus sampling, the preterm birth rate in pregnancies with

conned placental mosaicism was 45 percent, and 50 percent

o aected newborns were small or gestational age (outain,

2018). Placental mosaicism or trisomy 16 coners a particularly poor prognosis, with as ew as 1 in 3 pregnancies resulting

in normal outcome (Grau Madsen, 2018).

Gonadal Mosaicism

Also called germline mosaicism, this reers to having one cell

line in gametes and another in somatic cells. A genetic abnormality that is conned to gamete cells will aect all cells in the

ospring. Tus, gonadal mosaicism can account or apparently

de-novo diseases in the ospring o normal parents. Because

spermatogonia and oogonia divide throughout etal lie, and

spermatogonia continue to divide throughout adulthood, a

meiotic error can occur in germ cells that were previously normal. New mutations identied in an ospring whose ather is

older than 40 may arise through this mechanism, as discussed

later (p. 319) (Wilkie, 2017). Gonadal mosaicism also explains

the 6-percent recurrence risk ater the birth o a child with a

disease caused by a “new” mutation.

MODES OF INHERITANCE

A monogenic or mendelian disorder is caused by a mutation

or alteration in a single locus or gene in one or both members

o a gene pair. ypes o mendelian inheritance include autosomal dominant, autosomal recessive, X-linked, and Y-linked

(Table 16-3). Other monogenic inheritance patterns include

mitochondrial inheritance, uniparental disomy, imprinting,

and trinucleotide repeat expansion.

■ Relationship between Phenotype

and Genotype

When considering inheritance, it is the phenotype that is dominant or recessive, not the genotype. I a disease is dominant, a

normal gene directs the production o normal protein, but the

phenotype will be abnormal because o protein produced by

the abnormal gene. A heterozygous carrier o a recessive disease

may produce detectable levels o an abnormal gene product but

have no eatures o the condition, because the phenotype is

directed by the product o the normal co-gene. For example,

FIGURE 16-8 Pericentric inversion involves the centromere,

whereas paracentric inversion does not. Individuals with pericentric

inversions are at risk to have offspring with a duplication/deletion.

Those with paracentric inversions are at increased risk for early

pregnancy loss.

proximally than the telomere, the carrier is likely to be phenotypically abnormal. An example o this is the ring X chromosome, which may result in urner syndrome.

■ Mosaicism

A mosaic individual has two or more cytogenetically distinct

cell lines that are derived rom a single zygote. Phenotypic

expression o mosaicism depends on several actors, including

whether the cytogenetically abnormal cells involve the etus,

part o the etus, just the placenta, or some combination. Mosaicism is ound in approximately 0.3 percent o amnionic uid

cultures (Carey, 2014). When abnormal cells are present in

only a single ask o amnionic uid, the nding is likely pseudomosaicism, caused by cell-culture artiact (Bui, 1984; Hsu,

1984). When abnormal cells involve multiple cultures, however, true mosaicism is more likely. Further testing veries a

second cell line in 60 to 70 percent o these etuses (Hsu, 1984;

Worton, 1984).

Confined Placental Mosaicism

Mosaicism involving a gene abnormality is detected in 1 to 2

percent o chorionic villus sampling specimens. AmniocentesisGenetics 319

CHAPTER 16

erythrocytes rom carriers o sickle-cell anemia contain approximately 30 percent hemoglobin S, but because the other 70 percent is hemoglobin A, these cells do not usually sickle in vivo.

Heterogeneity

Genetic heterogeneity explains how dierent genetic mechanisms

may result in the same phenotype. Locus heterogeneity indicates

that a specic disease phenotype can be caused by mutations in

dierent genetic loci. It also explains why some diseases appear

to ollow more than one type o inheritance. For example, retinitis pigmentosa may develop ollowing mutations in at least 35

dierent genes or loci and may result in autosomal dominant,

autosomal recessive, or X-linked orms. Allelic heterogeneity

describes how dierent mutations o the same gene may aect

presentation o a particular disease. For example, although only

one gene has been associated with cystic brosis—the cystic

brosis conductance transmembrane regulator gene—more than

2000 mutations in this gene have been described and result in

variable disease severity (Chaps. 17, p. 342 and 54, p. 968).

Phenotypic heterogeneity explains how dierent disease states can

arise rom dierent mutations in the same gene. As an example, mutations in the broblast growth actor receptor 3 (FGFR3)

gene may result in several dierent skeletal disorders, including

achondroplasia and thanatophoric dysplasia, both o which are

discussed in Chapter 15 (p. 302).

■ Autosomal Dominant Inheritance

I only one copy o a gene pair determines the phenotype, that

gene is considered to be dominant. Carriers have a 50-percent

chance o passing on the aected gene with each conception.

A gene with a dominant mutation generally species the phenotype in preerence to the normal gene. Tat said, not all

individuals will necessarily maniest an autosomal dominant

condition the same way. Factors that aect the phenotype o

an autosomal dominant condition include penetrance, expressivity, and presence o codominant genes.

Penetrance

Tis characteristic describes whether a dominant gene is

expressed. A gene with recognizable phenotypic expression in

all individuals has complete penetrance. Penetrance is incomplete i some carriers express the gene but others do not. A gene

that is expressed in 80 percent o individuals is 80-percent penetrant. Incomplete penetrance explains why some autosomal

dominant diseases may appear to “skip” generations.

Expressivity

Individuals with the same autosomal dominant trait may maniest the condition dierently, even within the same amily.

Genes with variable expressivity can produce disease maniestations that range rom mild to severe. Examples include neurobromatosis, tuberous sclerosis, and adult polycystic kidney

disease.

Codominant Genes

I two dierent alleles in a gene pair are both expressed in the

phenotype, they are considered to be codominant. Blood type,

or example, is determined by expression o dominant A and B

red-cell antigens that can be expressed simultaneously. Another

example o codominance is the group o genes responsible or

hemoglobin production. I one gene directs production o

hemoglobin S and the other directs production o hemoglobin C, that individual will produce both S and C hemoglobin

(Chap. 59, p. 1053).

Advanced Paternal Age

Paternal age older than 40 is associated with increased risk

or spontaneous genetic mutations. Spermatogonia undergo

TABLE 16-3. Selected Monogenic (Mendelian) Disorders

Autosomal Dominant

Achondroplasia

Acute intermittent porphyria

Adult polycystic kidney disease

Antithrombin III deficiency

BRCA1 and BRCA2 breast and/or ovarian cancer

Ehlers-Danlos syndrome

Familial adenomatous polyposis

Familial hypercholesterolemia

Hereditary hemorrhagic telangiectasia

Hereditary spherocytosis

Huntington disease

Hypertrophic obstructive cardiomyopathy

Long QT syndrome

Marfan syndrome

Myotonic dystrophy

Neurofibromatosis type 1 and 2

Tuberous sclerosis

von Willebrand disease

Autosomal Recessive

α1-Antitrypsin deficiency

Congenital adrenal hyperplasia

Cystic fibrosis

Gaucher disease

Hemochromatosis

Homocystinuria

Phenylketonuria

Sickle-cell anemia

Tay-Sachs disease

Thalassemia syndromes

Wilson disease

X-Linked

Androgen insensitivity syndrome

Chronic granulomatous disease

Color blindness

Fabry disease

Fragile X syndrome

Glucose-6-phosphate deficiency

Hemophilia A and B

Hypophosphatemic rickets

Muscular dystrophy—Duchenne and Becker

Ocular albinism type 1 and 2320 The Fetal Patient

Section 6

mitotic division every 16 days, and the many replications raise

the risk or single base-pair mutations. As a result, ospring

o older athers are at risk or new autosomal dominant disorders and X-linked carrier states. Several in particular have

been termed paternal age efect disorders (Goriely 2012). Tese

include mutations in the broblast growth actor receptor 2

(FGFR2) gene, which may cause craniosynostosis syndromes

such as Apert, Crouzon, and Peier syndromes; mutations in

the FGFR3 gene, which may result in achondroplasia and thanatophoric dysplasia; and mutations in the RET proto- oncogene,

which may cause multiple endocrine neoplasia syndromes

(oriello, 2008).

Using whole genome sequencing to study SNPs among o-

spring o older athers, Kong and associates (2012) ound an

increase o approximately two mutations or each year o paternal age past 40. Te absolute risk or any specic condition

is low, because individual autosomal dominant disorders are

uncommon. Tus, no screening or testing is specically recommended.

■ Autosomal Recessive Inheritance

Recessive diseases develop only when both gene copies are

abnormal. Unless carriers are screened or a specic disease,

they usually are recognized only ater the birth o an aected

child or the diagnosis o an aected amily member. I a couple

has a child with an autosomal recessive disease, the recurrence

risk is 25 percent or each subsequent pregnancy. Tus, 1/4 o

ospring will be homozygous normal, 2/4 will be heterozygous

carriers, and 1/4 will be homozygous abnormal. In other words,

three o our children will be phenotypically normal, and 2/3 o

phenotypically normal siblings are actually carriers.

A heterozygous carrier o a recessive condition is only at risk

to have aected children i her or his partner is heterozygous

or homozygous or the disease. Genes or rare autosomal recessive conditions have low prevalence in the general population.

Tus, the likelihood that a partner will be a gene carrier is

small, unless there is consanguinity or the partner is a member

o an at-risk group. Carrier screening is discussed in Chapter 17

(p. 342). Preimplantation genetic testing allows a blastocyst to

be genetically evaluated prior to intrauterine transer during in

vitro ertilization and is described also in Chapter 17 (p. 348).

Inborn Errors of Metabolism

Most o these autosomal recessive diseases result rom absence

o a crucial enzyme, leading to incomplete metabolism o proteins, lipids, or carbohydrates. Te metabolic intermediates that

build up are toxic to various tissues and may result in intellectual disability or other abnormalities.

Phenylketonuria. Also known as phenylalanine hydroxylase

(PAH) deciency, this autosomal recessive disease is caused by

mutations in the PAH gene. PAH metabolizes phenylalanine to

tyrosine, and homozygotes have diminished or absent enzyme

activity. Tis leads to abnormally high levels o phenylalanine,

which results in progressive intellectual impairment, autism,

seizures, motor decits, and neuropsychological abnormalities (Blau, 2010). Because phenylalanine competitively inhibits

tyrosine hydroxylase, which is essential or melanin production,

aected individuals also have hair, eye, and skin hypopigmentation. More than 600 PAH gene mutations have been characterized, and the carrier requency varies by ethnicity. For those o

Northern European origin, carrier requency is 1 in 50, such

that the disease aects up to 1 in 10,000 newborns (American

College o Obstetricians and Gynecologists, 2020b). Prompt

diagnosis and restriction o dietary phenylalanine beginning

early in inancy are essential to prevent neurological damage.

All states mandate newborn screening or phenylketonuria

(PKU).

Phenylalanine restriction alone would result in inadequate

protein consumption, and phenylalanine-ree amino acid–

based supplementation is required. Maintenance o phenylalanine concentrations in the range o 2 to 6 mg/dL (120 to

360 μmol/L) is recommended rom at least 3 months prior

to pregnancy and continuing throughout pregnancy (American

College o Obstetricians and Gynecologists, 2020b).

Unortunately, phenylalanine is actively transported to the

etus. Women with PKU whose phenylalanine levels remain

above the recommended range during pregnancy are at risk to

have otherwise normal (heterozygous) ospring who sustain

signicant in utero damage as a result o being exposed to toxic

phenylalanine concentrations. Hyperphenylalaninemia raises

the risk or miscarriage and or PKU embryopathy, which is

characterized by intellectual disability, microcephaly, seizures,

growth impairment, and cardiac anomalies. Among women

on unrestricted diets, the risk to have a child with intellectual

disability exceeds 90 percent, microcephaly develops in more

than 70 percent, and as many as 1 in 6 children have cardiac

deects. Te Maternal Phenylketonuria Collaborative Study,

which included 572 pregnancies ollowed more than 18 years,

reported that maintenance o serum phenylalanine levels in the

recommended range signicantly reduced the etal abnormality

risk and resulted in normal childhood IQ scores (Koch, 2003;

Platt, 2000).

Approximately 25 to 50 percent o individuals with PKU

experience a signicant decline in phenylalanine levels when

treated with the synthetic PAH coactor tetrahydrobiopterin

(sapropterin) (Vockley, 2014). Based on registry data, sapropterin is considered a treatment option or those in whom

phenylalanine levels remain elevated despite dietary therapy

(Grange, 2014). Preconceptional counseling and consultation

with providers rom experienced PKU centers is recommended.

Consanguinity

In medical genetics, a union is consanguineous i between second cousins or closer relatives. Although uncommon in Western countries, the global estimate o consanguineous parentage

approximates 10 percent o the population (Oniya, 2019). More

than 1 billion people are estimated to live in countries in which

20 to 50 percent o marriages are consanguineous (Romeo,

2014). First-degree relatives share hal o their genes, seconddegree relatives share a ourth, and third-degree relatives—

rst cousins—share one eighth. Because o the potential or

shared deleterious genes, consanguinity coners an increased

risk to have ospring with otherwise rare autosomal recessive diseases or multiactorial disorders. In population-basedGenetics 321

CHAPTER 16

series, rst cousins have twice the risk or ospring with congenital abnormalities (Sheridan, 2013; Stoltenberg, 1997).

Consanguinity is also associated with a greater rate o stillbirth

(Kapurubandara, 2016). Because CMA perormed using a SNP

platorm may identiy consanguinity, it is important that preprocedural counseling include this possibility.

Incest is dened as a sexual relationship between rst-degree

relatives such as parent-child or brother-sister and is universally

illegal. Progeny o such unions carry the highest risk o abnormal outcomes. Older studies reported that up to 40 percent o

ospring were abnormal as a result o recessive and multiactorial disorders (Baird, 1982; Freire-Maia, 1984).

■ XLinked and YLinked Inheritance

Most X-linked diseases are recessive. Common examples

include color blindness, hemophilia A and B, and Duchenne

and Becker muscular dystrophy. A male with an X-linked

recessive gene is usually aected by the disease it causes,

because he lacks a second X chromosome to express the normal dominant gene. However, a male with an X-linked disease cannot have aected sons because they do not receive

his X chromosome. When a woman carries a gene causing an

X-linked recessive condition, each son has a 50-percent risk

o being aected, and each daughter has a 50-percent chance

o being a carrier.

Women with an X-linked recessive gene are generally una-

ected by the disease it causes. In some cases, however, the random inactivation o one X chromosome in each cell—termed

lyonization—is skewed, and emale carriers may have eatures

o the condition. For example, approximately 10 percent o

emale carriers o hemophilia A will have actor VIII levels less

than 30 percent o normal, and a similar proportion o emale

hemophilia B carriers have actor IX levels less than 30 percent.

Levels below these thresholds coner a greater risk or abnormal

bleeding when aected women give birth (Plug, 2006). Indeed,

even with higher levels, carriers are reported to be at increased

risk or bleeding complications (Olsson, 2014). Female carriers

o Duchenne or Becker muscular dystrophy carry an elevated

risk or cardiomyopathy, and periodic evaluation or cardiac

dysunction and neuromuscular disorders is recommended

(American Academy o Pediatrics, 2008).

X-linked dominant disorders mainly aect emales, because

they tend to be lethal in males. wo examples are vitamin D–

resistant rickets and incontinentia pigmenti. An important

exception is ragile X syndrome, discussed subsequently.

Te prevalence o Y-linked disorders is low. Te Y-chromosome carries genes important or sex determination and various

cellular unctions related to spermatogenesis and bone development. Deletion o genes on the long arm o Y results in severe

spermatogenic deects, whereas genes at the tip o the short

arm are critical or chromosomal pairing during meiosis and

or ertility.

■ Mitochondrial Inheritance

Human cells contain hundreds o mitochondria, each with

its own genome and associated replication system. Oocytes

have approximately 100,000 mitochondria. Sperm have only

about 100, and the latter are destroyed ater ertilization. Each

mitochondrion has multiple copies o a 16.5-kb circular DNA

molecule that contains 37 genes. Mitochondrial DNA encodes

peptides required or oxidative phosphorylation and encodes

ribosomal and transer RNAs.

Mitochondria are inherited exclusively rom the mother.

Tus, although males and emales both can be aected by a

mitochondrial disorder, a male cannot transmit the condition to his ospring. When a cell replicates, mitochondrial

DNA sorts randomly into each o the resulting cells, a process

termed replicative segregation. As a consequence o replicative

segregation, any mitochondrial mutation will be propagated

randomly into the newly ormed cells. Because each cell holds

multiple copies o mitochondrial DNA, the mitochondrion

may contain only normal or only abnormal DNA, termed

homoplasmy. Alternatively, it may contain both normal and

mutated DNA, namely heteroplasmy. I a heteroplasmic oocyte

is ertilized, the relative proportion o mutated DNA may

aect whether the individual maniests a given mitochondrial

disease. It is not possible to predict the potential degree o

heteroplasmy among ospring, and this poses challenges or

genetic counseling.

Tere are 33 mitochondrial diseases or conditions with

known molecular bases (Johns Hopkins University, 2020).

Examples include myoclonic epilepsy with ragged red bers

(MERRF), Leber optic atrophy, Kearns-Sayre syndrome, Leigh

syndrome, and several orms o mitochondrial myopathy and

cardiomyopathy.

■ DNA Triplet Repeat Expansion

Mendel's rst law is that genes are passed unchanged rom parent

to progeny, and barring new mutations, this is oten true. Certain

genes, however, contain a region o DNA trinucleotide repeats

that can expand during parent-to-child transmission. Te expansion can occur during male meiosis, as in Huntington disease,

or during emale meiosis, as in ragile X syndrome. Clinically,

DNA triplet repeat expansion is maniested by anticipation—a

phenomenon in which a disease may maniest at an earlier age or

become more severe with each successive generation. Examples

o DNA triplet repeat diseases are shown in Table 16-4.

Fragile X Syndrome

Tis is the most common inherited intellectual disability. Fragile X syndrome aects approximately 1 in 3600 males and 1 in

TABLE 16-4. Some Disorders Caused by DNA Triplet

Repeat Expansion

Dentatorubral-pallidoluysian atrophy

Fragile X syndrome

Friedreich ataxia

Huntington disease

Spinal and bulbar muscular atrophy

Myotonic dystrophies

Spinocerebellar ataxias322 The Fetal Patient

Section 6

4000 to 6000 emales (American College o Obstetricians and

Gynecologists, 2020a). It is caused by expansion o a repeated trinucleotide DNA segment—cytosine-guanine-guanine (CGG)—

at chromosome Xq27.3. When the CGG repeat number reaches

a critical size—the ull mutation—the ragile X mental retardation 1 (FMR1) gene becomes methylated and inactivated. Inactivation halts expression o the FMR1 protein, which is abundant

in nerve cells and necessary or normal cognitive development.

Individuals with ragile X syndrome may have speech and

language problems, attention-decit/hyperactivity disorder,

and autism or autistic-like behaviors. Intellectual disability is

generally more severe in males, in whom average IQ scores

are 35 to 45 (Nelson, 1995). Te syndrome has characteristic

phenotypic eatures—a narrow ace with large jaw, prominent

ears, connective tissue abnormalities, and macroorchidism in

postpubertal males.

Both the sex o the aected individual and the number o

CGG repeats determine the degree o clinical impairment.

Clinically, our groups have been described (American College

o Obstetricians and Gynecologists, 2020a):

• Full mutation—more than 200 repeats

• Premutation—55 to 200 repeats

• Intermediate—45 to 54 repeats

• Unaected—fewer than 45 repeats.

Full mutations are expressed in all males and many emales.

Males with ull mutations typically have signicant cognitive and behavioral abnormalities and phenotypic eatures. In

emales, random X chromosome inactivation results in variable

expression, and the disability may be much less severe. With

rare exception, the parent o origin o repeat expansion that

leads to a ull mutation is emale (Monaghan, 2013).

Counseling or a premutation is more complex. Te woman

with a ragile X premutation may have ospring with the ull

mutation, and the risk ranges rom ≤5 percent with ewer than

70 CGG repeats to >95 percent with 100 to 200 CGG repeats

(Nolin, 2003). Expansion is extremely unlikely in a male premutation carrier, but all o his daughters will carry the premutation. Among women with no risk actors, approximately 1 in

250 carries a ragile X premutation, and in those with a amily

history o intellectual disability, the risk approximates 1 in 90

(Cronister, 2008).

Fragile X premutation carriers may themselves experience

signicant health consequences. Males with a premutation

are at risk or the ragile X tremor ataxia syndrome (FXAS),

which is characterized by memory loss, executive unction de-

cits, anxiety, and dementia (Monaghan, 2013). Females are at

decreased risk or FXAS, but they have a 20-percent risk or

ragile X-associated primary ovarian insufciency (POI).

Carrier screening is recommended or women with a amily history o ragile X syndrome, or those with unexplained

intellectual disability, developmental delay, or autism, and

or women with POI (American College o Obstetricians

and Gynecologists, 2020a). Prenatal diagnosis can be accomplished by amniocentesis or chorionic villus sampling. Either

type o specimen can be used to evaluate the CGG repeat number, but chorionic villus sampling may not accurately determine

FMR1 gene methylation status.

■ Imprinting

Usually, one copy o each gene is inherited rom each parent,

and both copies are expressed. Imprinting reers to the situation in which gene expression varies according to the parent

o origin. With maternal imprinting, a gene inherited rom the

mother is transcriptionally silent and the gene inherited rom

the ather is active. Paternal imprinting is the opposite. Tis

phenomenon explains the phenotypic dierences that occur in

triploid pregnancies according to whether the extra chromosomal set is maternal or paternal (p. 313).

Imprinting aects gene expression by epigenetic control,

that is, it modies the phenotype without altering the underlying genetic structure or genotype. Importantly, the eect may

be reversed in a subsequent generation, because a emale who

inherits an imprinted gene rom her ather will pass it in her

oocytes with a maternal—rather than paternal—imprint, and

vice versa.

An example o imprinting can be ound in two very dierent diseases associated with the same region o DNA. PraderWilli syndrome is characterized by pronounced hyperphagia

with obesity, along with short stature, small hands and eet,

and mild mental retardation. In more than 70 percent o cases,

Prader-Willi is caused by microdeletion or disruption o a

DNA segment on the paternal chromosome 15, 15q11.2-q13.

Te remaining cases are due to maternal uniparental disomy

or due to maternal gene imprinting with the paternal gene

inactivated.

In contrast, individuals with Angelman syndrome have normal stature and weight but severe intellectual disability, absent

speech, seizures, ataxia, jerky arm movements, and paroxysms

o inappropriate laughter. In approximately 70 percent o

cases, Angelman syndrome is caused by microdeletion or the

15q11.2-q13 DNA segment on the mother's chromosome 15.

Tis segment contains a gene that codes or ubiquitin protein

ligase E3A, an important neural protein. Paternal gene imprinting deects—with maternal genes inactivated—account or 2

to 3 percent, and 2 percent result rom paternal uniparental

disomy. Table 16-5 lists selected diseases that can involve

imprinting.

■ Uniparental Disomy

Tis occurs when both members o a chromosome pair are

inherited rom the same parent. Uniparental disomy usually

TABLE 16-5. Some Disorders That Can Involve Imprinting

Disorder

Chromosomal

Region

Parental

Origin

Angelman 15q11.2–q13 Maternal

Beckwith-Wiedemann 11p15.5 Paternal

Myoclonus-dystonia 7q21 Maternal

Prader-Willi 15q11.2–q13 Paternal

Pseudohypoparathyroidism 20q13.2 Variable

Russell-Silver syndrome 7p11.2 Maternal

Data from Johns Hopkins University, 2021.Genetics 323

CHAPTER 16

has no clinical consequences, because gene expression is not

generally aected by the parent o origin and because both copies o a chromosome pair are not usually identical. However,

i chromosomes 6, 7, 11, 14, or 15 are involved, ospring are

at increased risk or an abnormality because o parent-o-origin

dierences in gene expression, that is, because o imprinting

(Shaer, 2001). Several genetic mechanisms may cause uniparental disomy, the most common o which is trisomic rescue,

shown in Figure 16-9. Ater a nondisjunction event produces

a trisomic conceptus, one o the three homologues may be

lost, resulting in uniparental disomy or that chromosome in

approximately one third o cases.

Uniparental isodisomy occurs when an individual receives

two identical copies o one chromosome rom the same parent.

Tis mechanism has explained cases o cystic brosis in which

only one parent is a carrier but the etus inherits two copies o

the same gene mutation (Spence, 1988; Spotila, 1992).

■ Multifactorial Inheritance

raits or diseases that are determined by the combination o

multiple genes and environmental actors are considered to

have multiactorial inheritance (Table 16-6). Polygenic traits

are determined by the combined eects o more than one gene.

Many congenital and acquired conditions and common traits

display multiactorial inheritance. Examples include malormations such as neural-tube deects, cardiac deects, and acial

clets; diseases such as diabetes and atherosclerosis; and traits

such as head size or height. Multiactorial abnormalities tend

to recur in amilies, but not according to a mendelian pattern.

I a couple has had 1 child with a multiactorial birth deect,

the risk that another child will be aected is empirically 3 to 5

percent. Tis risk declines exponentially with successively more

distant amily relationships.

Multiactorial traits that have a normal distribution in the

population are termed continuously variable. A measurement that

is more than two standard deviations above or below the population mean may be considered abnormal. Continuously variable

traits tend to be less extreme in the ospring o aected individuals, because o the statistical principle o regression to the mean.

TABLE 16-6. Characteristics of Multifactorial Diseases

There is a genetic contribution

No mendelian pattern of inheritance

No evidence of single-gene disorder

Nongenetic factors are also involved in disease

causation

Lack of penetrance despite predisposing genotype

Monozygotic twins may be discordant

Familial aggregation may occur

Relatives are more likely to have disease-predisposing

alleles

Expression more common among close relatives

Greater concordance in monozygotic than dizygotic

twins

Becomes less common in less closely related relatives—

fewer predisposing alleles

Adapted from Nussbaum, 2016.

Normal Uniparental

disomy

Normal

A B

FIGURE 16-9 Mechanism of uniparental disomy arising from trisomic “rescue.” A. In normal meiosis, one member of each pair of homologous chromosomes is inherited from each parent. B. If nondisjunction results in a trisomic conceptus, one homologue is sometimes lost. In

a third of cases, loss of one homologue leads to uniparental disomy.324 The Fetal Patient

Section 6

Threshold Traits

Some multiactorial traits do not appear until a threshold is

exceeded. Genetic and environmental actors that create propensity or liability or the trait are themselves normally distributed, and only individuals at extremes o the distribution

exceed the threshold and exhibit the trait or deect. Te phenotypic abnormality is thus an all-or-none phenomenon.

Certain threshold traits have a clear male or emale predominance. I an individual o the less aected gender has the

characteristic or deect, the risk is greater in his or her ospring

(Fig. 16-10). An example is pyloric stenosis, which is approximately our times more common in males (Krogh, 2012). A

emale with pyloric stenosis has likely inherited more predisposing genetic actors than are necessary to produce the deect

in a male, and the recurrence risk or her children or siblings is

thus higher than the expected 3 to 5 percent. Her brothers and

sons would have the highest liability because they not only will

inherit more than the usual number o predisposing genes but

also are the more susceptible gender.

Te recurrence risk or threshold traits is also greater i the

deect is severe. For example, the recurrence risk ater the birth

o a child with bilateral clet lip and palate approximates 8 percent, but it is only about 4 percent ollowing a child with unilateral clet lip alone.

Cardiac Defects

Structural cardiac anomalies are the most common birth

deects, with a birth prevalence o 8 cases per 1000. Te risk o

having a child with a cardiac anomaly approximates 5 to 6 percent i the mother has the deect and 2 to 3 percent i the ather

is aected (Burn, 1998). Selected let-sided lesions, including

hypoplastic let heart syndrome, coarctation o the aorta, and

bicuspid aortic valve, may have recurrence risks our- to six-

old higher (Lin, 1988; Lupton, 2002; Nora, 1988). Observed

recurrence risks or specic cardiac malormations are listed in

able 52-4 (p. 920).

Neural-tube Defects

Risk or developing a neural-tube deect (ND) is inuenced by

amily history, hyperthermia, hyperglycemia, teratogen exposure, obesity, ethnicity, and etal gender. More than 50 years

ago, Hibbard and Smithells (1965) postulated that abnormal

olate metabolism was responsible or many NDs. However,

most ND cases do not occur in the setting o maternal olic

acid deciency, and it has become clear that the gene-nutrient

interactions underlying olate-responsive NDs are complex.

For a woman with a prior aected child, the 3- to 5-percent

recurrence risk declines by at least 70 percent with 4 mg/d

o periconceptional oral olic acid supplementation (Grosse,

2007; MRC Vitamin Study Research Group, 1991). Periconceptional olic acid supplementation also appears to ameliorate

the etal ND risk in women with pregestational diabetes and

in those with exposure to antiepileptic medications, ever during embryogenesis, and obesity (Kerr, 2017; Petersen, 2019).

Sonographic eatures o NDs are described in Chapter 15

(p. 276), their prevention with olic acid is discussed in Chapter

9 (p. 168), and etal therapy or myelomeningocele is reviewed

in Chapter 19 (p. 372).

GENETIC TESTS

All pregnant women should have the option o prenatal aneuploidy screening and prenatal genetic diagnosis (American College o Obstetricians and Gynecologists, 2018). Screening or

etal aneuploidy may be either serum analyte-based or cell-ree

DNA-based. Prenatal genetic carrier screening tests or cystic brosis and spinal muscular atrophy carrier status also are

routinely oered. Additional carrier screening tests are oered

to at-risk individuals. Expanded genetic carrier screening panels also are available. Te American College o Obstetricians

and Gynecologists considers ethnic-specic, panethnic, and

expanded carrier screening acceptable strategies (American College o Obstetricians and Gynecologists, 2020a). Tese screening tests are discussed in Chapter 17 (p. 342).

For prenatal genetic diagnosis, the most commonly used

tests are chromosomal microarray analysis (CMA), cytogenetic

analysis (karyotyping), and uorescence in situ hybridization

(FISH). esting may be perormed on amnionic uid or chorionic villi. In selected circumstances, whole genome or whole

exome sequencing may be considered, but these are not recommended or routine use. o diagnose a specic disease whose

genetic basis is known, DNA-based tests are oten employed,

typically using polymerase chain reaction (PCR) or rapid

amplication o DNA sequences.

■ Cytogenetic Analysis

Also known as karyotyping, cytogenetic analysis tests or

numerical chromosomal abnormalities—aneuploidy or polyploidy. It can also identiy balanced or unbalanced structural

rearrangements o at least 5 to 10 megabases in size. Karyotyping has diagnostic accuracy exceeding 99 percent.

Any tissue containing dividing cells or cells that can be stimulated to divide is suitable or cytogenetic analysis. Te dividing cells are arrested in metaphase, and their chromosomes are

stained to reveal light and dark bands. Te most commonly

used technique is Giemsa staining, which yields the G-bands

shown in Figure 16-3. Each chromosome has a unique banding pattern that permits its identication and detection o

Unaffected Affected

THRESHOLD

Males

Females

Population

Trait Liability

FIGURE 16-10 Schematic example of a threshold trait, such as

pyloric stenosis, which has a predilection for males. Each gender is

normally distributed, but at the same threshold, more males than

females will develop the condition.Genetics 325

CHAPTER 16

deleted, duplicated, or rearranged segments. Te accuracy o

cytogenetic analysis rises with the number o bands produced.

Metaphase banding routinely yields 450 to 550 visible bands

per haploid chromosome set, whereas prophase banding generally yields 850 bands.

Only dividing cells can be evaluated, so the growth o cultured cells aects the rapidity with which results are obtained.

Amnionic uid, which contains amniocytes, epithelial cells,

and gastrointestinal mucosal cells, will usually yield results in

7 to 10 days (Chap. 17, p. 344). Fetal blood cells may provide results in 36 to 48 hours but are rarely needed. I etal

skin broblasts are evaluated postmortem, stimulation o cell

growth can be more difcult. Cytogenetic analysis may take 2

to 3 weeks in such cases and has largely been replaced by CMA

(Chap. 35, p. 627).

■ Fluorescence In Situ Hybridization

Tis technique is most commonly used or rapid identication

o a specic chromosomal abnormality, such as trisomy 21, 18,

or 13. Because o its 1- to 2-day turnaround time, FISH is typically selected or cases in which ndings may alter pregnancy

management. It may also be used to veriy a suspected deletion syndrome, such as the 22q11.2 microdeletion (p. 315),

although CMA is preerred. Te American College o Obstetricians and Gynecologists (2018) recommends that decisionmaking based on FISH should incorporate clinical inormation

consistent with the suspected diagnosis. Supportive data may

be an abnormal aneuploidy screening test result, a sonographic

nding, or a conrmatory diagnostic test such as karyotyping

or CMA.

o perorm FISH, cells are xed onto a glass slide, and

uorescent-labeled probes are hybridized to the xed chromosomes (Fig. 16-11). Each probe is a DNA sequence that is complementary to a chromosome region or gene being investigated.

I the DNA sequence is present, hybridization is detected as a

bright signal visible by microscopy. Te number o signals indicates the number o chromosomes or genes o that type in the

cell. FISH does not provide inormation on the entire chromosomal complement but merely the chromosomal or gene region

o interest. Figure 16-12 shows an example o interphase FISH

using α-satellite probes or chromosomes 18, X, and Y to con-

rm trisomy 18.

■ Chromosomal Microarray Analysis

Tis test is 100 times more sensitive than karyotyping and

detects microduplications and microdeletions as small as 50

to 100 kilobases. Direct CMA, which is perormed on uncultured cells, can yield results in 3 to 7 days. I cultured cells are

required, results may take 10 to 14 days (American College o

Obstetricians and Gynecologists, 2018).

DNA

probe

DNA probe

labeled with

fluorescent dye

DNA denatured

and separated

Probe creation

Probe hybridized with

patient chromosomes

Fluorescent probe

illuminates chromosome

region of interest

FIGURE 16-11 Steps in fluorescence in situ hybridization (FISH).

Interphase FISH

Y Chromosome = Red

18 Chromosome = Light blue

X Chromosome = Green

FIGURE 16-12 Interphase fluorescence in situ hybridization (FISH)

using α-satellite probes for chromosomes 18, X, and Y. In this case,

the three light blue signals, two green signals, and absence of red

signals indicate that this is a female fetus with trisomy 18. (Reproduced with permission from Dr. Frederick Elder.)326 The Fetal Patient

Section 6

Microarrays use either a comparative genomic hybridization (CGH) platorm, a SNP platorm, or a combination o

the two. Te CGH microarray platorm compares specimen

DNA with a normal control sample. Shown in Figure 16-13,

the CGH chip contains reerence DNA ragments o known

sequence—oligonucleotides. Fetal DNA rom the amniocentesis or chorionic villus sampling specimen is labeled with a

uorescent dye and then hybridized to the DNA on the chip.

Normal control DNA is labeled with a dierent probe and also

hybridized to the same chip. Te intensity o the uorescent

signals rom the two samples is compared. With a SNP array,

the chip contains known DNA sequence variants, that is, singlenucleotide polymorphisms. When etal DNA is labeled and

hybridized to the chip, the uorescent signal intensity indicates

copy number variation.

Both types o platorms detect aneuploidy, unbalanced

translocations, and microdeletions and microduplications. In

addition, SNP arrays are able to identiy triploidy and can

detect absence o heterozygosity. Te latter can occur with uniparental disomy, in which both copies o a chromosome are

inherited rom one parent. Absence o heterozygosity may

also occur with consanguinity, and counseling prior to perormance o an SNP array should include this possibility (p. 321).

Importantly, neither type o array platorm currently detects

balanced chromosomal rearrangements. For this reason, couples

with recurrent pregnancy loss should be oered karyotyping as

the rst-line test (Society or Maternal-Fetal Medicine, 2016).

In addition to identiying pathogenic copy number variants, CMA may detect variants that are unable to be classied as

benign or pathologic and thus are considered to be o uncertain

clinical signicance. In recent series, these variants o uncertain

signicance are identied in approximately 1 percent o prenatal

specimens (Brady, 2014; Chong, 2019; Wang, 2018). Not unexpectedly, variants o uncertain signicance may be a source o

distress to amilies, even with comprehensive pretest counseling.

Clinical Applications

In pregnancies at increased risk or autosomal trisomy based

on aneuploidy screening results, karyotyping or FISH plus

karyotyping should be oered, and CMA should be made

available (American College o Obstetricians and Gynecologists, 2018). Clinicians are increasingly oering CMA in

this setting. I the karyotype is normal, CMA has identied

clinically relevant copy number variants in approximately 6.5

percent o pregnancies with etal abnormalities and in 1 to 2

percent o those without obvious etal abnormality (Callaway,

2013; Chong, 2019). Te American College o Obstetricians

and Gynecologists (2018) and the Society or Maternal-Fetal

Medicine (2016) recommend CMA as the rst-tier test when

etal structural abnormalities are identied. I a particular

1.28 cm

1.28 cm

Actual size of chip

500,000 cells on each chip

One cell on chip

Nonhybridized

DNA

Hybridized DNA

A

B

C

D E

F

Thousands of identical nucleotide

strands on one cell

Labeled fetal DNA is presented

to the cells

FIGURE 16-13 Chromosomal microarray analysis. A. Actual microarray chip size. B. Each chip contains thousands of cells (squares).

C & D. Each cell contains thousands of identical oligonucleotides on its surface and is unique in its nucleotide content. E. During genetic

analysis, a mixture containing tagged fetal DNA is presented to the chip. Complementary DNA sequences bind. F. If a laser is shined on the

chip, DNA sequences that have bound will glow thus identifying a matching sequence. (Modified with permission from Doody KJ: Treatment

of the infertile couple. In Hoffman BL, Schorge JO, Schaffer JI, et al (eds): Williams Gynecology, 2nd ed. New York, NY: McGraw Hill; 2012.)Genetics 327

CHAPTER 16

anomaly that strongly suggests a specic aneuploidy is identied, such as an endocardial cushion deect (trisomy 21)

or alobar holoprosencephaly (trisomy 13), then karyotyping

or FISH may be oered as the initial test. Genetic counseling should include inormation regarding the benets and

limitations o both CMA and karyotyping. Each should be

made available to women who elect prenatal diagnosis (Society or Maternal-Fetal Medicine, 2016). CMA may identiy

instances o autosomal dominant genetic disorders that have

not yet maniested in an aected parent, and it may also

identiy instances o nonpaternity.

For stillbirth evaluation, CMA is signicantly more likely

than standard karyotyping to provide a genetic diagnosis, in

part because it does not require dividing cells. Te Stillbirth

Collaborative Research Network ound that when karyotyping

was uninormative, approximately 6 percent o cases had either

aneuploidy or a pathogenic copy number variant identied

with CMA (Reddy, 2012). In a recent metaanalysis o more

than 900 stillbirths with normal karyotype, CMA identied

pathogenic copy number variants in 6 percent with structural

abnormalities and 3 percent with no abnormalities evident

(Martinez-Portilla, 2019). Overall, CMA yielded results 15

percent more oten than karyotyping alone.

■ Whole Genome and Whole Exome

Sequencing

Most etuses with structural abnormalities have a normal

karyotype and a normal CMA result. Whole genome sequencing (WGS) is a technique or analyzing the entire genome.

Whole exome sequencing (WES) analyzes only the DNA coding regions, which account or approximately 1.5 percent o the

genome. Tese next-generation sequencing tools are increasingly used postnatally to evaluate suspected genetic syndromes

and intellectual disability.

Several prospective, multicenter series have evaluated WES

or pregnancies in which etuses have structural abnormalities,

no karyotypic evidence o aneuploidy, and normal CMA. WES

identied genetic abnormalities in approximately 10 percent

o such cases (Lord, 2019; Petrovski, 2019). Utility was higher

in cases with cardiac, skeletal, or multiorgan system anomalies.

Importantly, WGS and WES have signicant limitations in

their current orm. Tese include prohibitively long turnaround

times, high costs, and increased rates o alse-positive and alsenegative results and variants o uncertain signicance (American

College o Medical Genetics, 2012; Atwal, 2014). In addition,

because sequencing is generally perormed on the etus and parents simultaneously, one parent may be identied or suspected

to have an unrelated but medically actionable nding, which

urther complicates counseling (American College o Obstetricians and Gynecologists, 2020c). As a result, the clinical utility

o this promising technology or prenatal cases is currently limited, and it is not recommended or routine clinical use.

■ Fetal DNA in the Maternal Circulation

Fetal cells are present in maternal blood at a very low concentration, only 2 to 6 cells per milliliter (Bianchi, 2006). Intact

etal cells sometimes persist in the maternal circulation or

decades ollowing delivery. Persistent etal cells may engrat in

the mother, causing microchimerism (Chap. 62, p. 1109). Tis

has been implicated in maternal autoimmune diseases such as

scleroderma, systemic lupus erythematosus, and Hashimoto

thyroiditis (Kinder, 2017). From the standpoint o prenatal

diagnosis, use o intact etal cells rom maternal blood is limited by low cell concentration, cell persistence into successive

pregnancies, and difculties in distinguishing etal and maternal

cells. Cell-ree DNA overcomes these limitations.

Cell-free DNA

Also known as cell-ree etal DNA or noninvasive prenatal

screening (NIPS), this screening test uses DNA ragments

derived rom maternal cells and rom apoptotic placental trophoblast cells. “Fetal” is thus a misnomer. Cell-ree DNA is

reliably detected in maternal blood ater 9 to 10 weeks’ gestation (Bianchi, 2018). Te proportion o total cell-ree DNA

that is placental is called the etal raction, and it accounts or

approximately 10 percent o the total circulating cell-ree DNA

in maternal plasma. Unlike intact etal cells, cell-ree DNA is

cleared within minutes rom maternal blood.

Current clinical applications o cell-ree DNA are aneuploidy screening and Rh D genotyping (Fig. 16-14). Commercial laboratories have also oered two other applications

o cell-ree DNA: (1) screening or selected microdeletion and

microduplication syndromes and (2) genome-wide screening

or deletions and duplications larger than 7 megabase pairs.

Currently, prospective data on these promising applications

are limited, and neither is recommended or routine screening

(American College o Obstetricians and Gynecologists, 2020c).

Additional applications or cell-ree DNA are on the horizon. In research settings, numerous single-gene disorders have

been detected and include skeletal dysplasias, hemoglobinopathies, cystic brosis, congenital adrenal hyperplasia, hemophilia,

muscular dystrophy, myotonic dystrophy, and spinal muscular

atrophy (Camunas-Soler, 2018; Jenkins, 2018; Zhang, 2019).

Aneuploidy Screening. Several dierent assay platorms are

used to screen or etal autosomal trisomies and sex chromosomal aneuploidies. Tese include massively parallel sequencing,

which is a WGS technique; chromosome-selective or targeted

sequencing; and analysis o SNPs (American College o Obstetricians and Gynecologists, 2018, 2020c). By simultaneously

sequencing millions o chromosome-specic DNA ragments,

investigators can identiy whether the proportion or ratio o

ragments rom one chromosome is higher than expected. Tus,

samples rom women with a Down syndrome etus will have

a larger proportion o DNA sequences rom chromosome 21.

Te screening perormance o cell-ree DNA is excellent. In

a metaanalysis o 35 studies o largely high-risk pregnancies,

the pooled sensitivity to detect Down syndrome was 99.7 percent, and to identiy trisomies 18 and 13, 98 and 99 percent,

respectively (Gil, 2017). For each, the specicity was at least

99.9 percent, such that the cumulative alse-positive rate was

below 1 percent. Aneuploidy screening reports commonly include

inormation about etal sex, which may be clinically useul i

the etus is at risk or an X-linked disorder or or congenital

adrenal hyperplasia (Chap. 17, p. 335). In a metaanalysis, the328 The Fetal Patient

Section 6

sensitivity o cell-ree DNA testing or etal sex determination

was reported to be 95 percent between 7 and 12 weeks’ gestation and 99 percent ater 20 weeks (Devaney, 2011).

Causes o alse-positive and alse-negative screening results are

listed in Table 16-7. As with any other screening test, diagnostic conrmation should be perormed beore irreversible medical

intervention. Cell-ree DNA screens do not yield a result in 2 to

4 percent o cases. Tis may be due to assay ailure, high assay

variance, or low etal raction (Norton, 2012; Pergament, 2014;

Quezada, 2015). Importantly, such pregnancies carry a greater

risk or etal aneuploidy. In addition, results may not reect the

etal DNA complement but rather may indicate conned placental mosaicism, early demise o an aneuploid co-twin, maternal mosaicism, or rarely, occult maternal malignancy (Bianchi,

2015; Curnow, 2015; Grati, 2014b; Wang, 2014). Recommendations or counseling are discussed in Chapter 17 (p. 336).

Rh D Genotype Evaluation. Many etuses o Rh D-negative

women are also Rh D-negative. In a predominantly white

population, this prevalence approaches 40 percent. Fetal Rh D

genotype assessment rom maternal blood can eliminate administration o anti-D immune globulin in such cases, thereby

reducing costs and potential risk. When Rh D alloimmunization is suspected, early identication o an Rh D-negative etus

might avoid unnecessary middle cerebral artery Doppler assessment or amniocentesis. Evaluation using cell-ree DNA is done

using real-time PCR to target several exons o the RHD gene,

typically exons 4, 5, 7, and 10.

Rh D-genotyping is perormed routinely with cell-ree

DNA in Denmark, Finland, and the Netherlands (Clausen,

2012; de Haas, 2016; Haimila, 2017). In population-based

studies o more than 35,000 Rh D-negative women screened

at 24 to 27 weeks’ gestation, the alse-negative rate—in which

Rh D-negative status was missed—was only 0.03 percent.

Te alse-positive rate—in which Rh immune globulin would

be given unnecessarily—was less than 1 percent (de Haas,

2016; Haimila, 2017). Similar results were reported rom the

United Kingdom, although the alse-negative rate was higher

in the rst trimester. Investigators concluded that alse-negative screening results might increase the alloimmunization

risk but by less than 1 case per million births (Chitty, 2014).

Rh D alloimmunization is discussed in Chapter 18 (p. 353).

TABLE 16-7. Selected Etiologies Creating Inaccurate

Cell-free DNA Results

Abnormality not detected—false negative

Low fetal fraction

Fetal chromosomal abnormality

Multifetal gestation

Maternal obesity

Low-molecular-weight heparin

Confined placental mosaicism (normal placenta,

aneuploid fetus)

Normal fetus but abnormal screen—false positive

Confined placental mosaicism (normal fetus, aneuploid

placenta)

Multifetal gestation with loss of aneuploid fetus

Maternal aneuploidy (e.g. 47,XXX)

Maternal mosaicism (e.g. 46,XX/45,X) or fetal mosaicism

Maternal cancer (lymphoma, breast, colon, leukemia,

others)

Vitamin B12 deficiency

Data from Bianchi, 2018.

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