Medical Genetics
19 基因治疗
Gene Therapy
Medical Genetics
1,What is gene therapy?
Genes,which are carried on
chromosomes,are the basic physical
and functional units of heredity,
Genes are specific sequences of
bases that encode instructions on
how to make proteins,
Medical Genetics
Although genes get a lot of
attention,it’s the proteins that
perform most life functions and even
make up the majority of cellular
structures,
Medical Genetics
When genes are altered so that
the encoded proteins are unable to
carry out their normal functions,
genetic disorders can result,
Medical Genetics
Gene therapy is a technique for
correcting defective genes
responsible for disease development,
Medical Genetics
Researchers may use one of several approaches
for correcting faulty genes,
1,A normal gene may be inserted into a nonspecific
location within the genome to replace a
nonfunctional gene,This approach is most common,
2,An abnormal gene could be swapped for a normal
gene through homologous recombination,
3,The abnormal gene could be repaired through
selective reverse mutation,which returns the gene
to its normal function,
4,The regulation (the degree to which a gene is turned
on or off) of a particular gene could be altered,
Medical Genetics
2,How does gene therapy work?
In most gene therapy studies,a
"normal" gene is inserted into the
genome to replace an "abnormal,"
disease-causing gene,
Medical Genetics
A carrier molecule called a
vector must be used to deliver the
therapeutic gene to the patient's
target cells,Currently,the most
common vector is a virus that has
been genetically altered to carry
normal human DNA,
Medical Genetics
Viruses have evolved a way of
encapsulating and delivering their
genes to human cells in a pathogenic
manner,
Medical Genetics
Scientists have tried to take
advantage of this capability and
manipulate the virus genome to
remove disease-causing genes and
insert therapeutic genes,
Medical Genetics
Target cells such as the patient's
liver or lung cells are infected with
the viral vector,The vector then
unloads its genetic material
containing the therapeutic human
gene into the target cell,
Medical Genetics
The generation of a functional
protein product from the therapeutic
gene restores the target cell to a
normal state,
Medical Genetics
To reverse disease caused by genetic damage,researchers isolate
normal DNA and package it into a vector,a molecular delivery truck
usually made from a disabled virus,Doctors then infect a target
cell — usually from a tissue affected by the illness,such as liver or
lung cells— with the vector,The vector unloads its DNA cargo,which
then begins producing the missing protein and restores the cell to
normal,
Medical Genetics
A,Some of the different types of
viruses used as gene therapy vectors
(1) Retroviruses
A class of viruses that can create
double-stranded DNA copies of their RNA
genomes,These copies of its genome can
be integrated into the chromosomes of
host cells,Human immunodeficiency virus
(HIV) is a retrovirus,
Medical Genetics
(2)Adenoviruses
A class of viruses with double-stranded DNA
genomes that cause respiratory,intestinal,and
eye infections in humans,The virus that causes
the common cold is an adenovirus,
(3)Adeno-associated viruses
A class of small,single-stranded DNA viruses
that can insert their genetic material at a specific
site on chromosome 19,
(4)Herpes simplex viruses
A class of double-stranded DNA viruses that
infect a particular cell type,neurons,Herpes
simplex virus type 1 is a common human pathogen
that causes cold sores,
Medical Genetics
Medical Genetics
Medical Genetics
Medical Genetics
Besides virus-mediated gene-
delivery systems,there are several
nonviral options for gene delivery,
The simplest method is the direct
introduction of therapeutic DNA into
target cells,This approach is limited
in its application because it can be
used only with certain tissues and
requires large amounts of DNA,
Medical Genetics
Another nonviral approach
involves the creation of an artificial
lipid sphere with an aqueous core,
This liposome,which carries the
therapeutic DNA,is capable of
passing the DNA through the target
cell's membrane,
Medical Genetics
Therapeutic DNA also can get
inside target cells by chemically
linking the DNA to a molecule that
will bind to special cell receptors,
Medical Genetics
Once bound to these receptors,
the therapeutic DNA constructs are
engulfed by the cell membrane and
passed into the interior of the target
cell,This delivery system tends to be
less effective than other options,
Medical Genetics
Researchers also are
experimenting with introducing a
47th artificial human chromosome
into target cells,This chromosome
would exist autonomously alongside
the standard 46 --not affecting their
workings or causing any mutations,
Medical Genetics
It would be a large vector capable of
carrying substantial amounts of genetic
code,and scientists anticipate that,
because of its construction and autonomy,
the body's immune systems would not
attack it,A problem with this potential
method is the difficulty in delivering such
a large molecule to the nucleus of a target
cell,
Medical Genetics
3,The current status of gene therapy research
Current gene therapy is experimental and
has not proven very successful in clinical trials,
Medical Genetics
Little progress has been made
since the first gene therapy clinical
trial began in 1990,In 1999,gene
therapy suffered a major setback
with the death of 18-year-old Jesse
Gelsinger,
Medical Genetics
Jesse was participating in a gene
therapy trial for ornithine
transcarboxylase deficiency (OTCD),
He died from multiple organ failures
4 days after starting the treatment,
His death is believed to have been
triggered by a severe immune
response to the adenovirus carrier,
Medical Genetics
Another major blow came in
January 2003,when the FDA (USA)
placed a temporary halt on all gene
therapy trials using retroviral vectors
in blood stem cells,
Medical Genetics
FDA (USA) took this action after
it learned that a second child treated
in a French gene therapy trial had
developed a leukemia-like condition,
Medical Genetics
Both this child and another who
had developed a similar condition in
August 2002 had been successfully
treated by gene therapy for X-linked
severe combined immunodeficiency
disease (X-SCID),also known as
"bubble baby syndrome."
Medical Genetics
FDA's Biological Response Modifiers
Advisory Committee (BRMAC) met at the
end of February 2003 to discuss possible
measures that could allow a number of
retroviral gene therapy trials for treatment
of life-threatening diseases to proceed
with appropriate safeguards,FDA has yet
to make a decision based on the
discussions and advice of the BRMAC
meeting,
Medical Genetics
4,Factors kept gene therapy effective
A,Short-lived nature of gene therapy
Before gene therapy can become a
permanent cure for any condition,the
therapeutic DNA introduced into target
cells must remain functional and the
cells containing the therapeutic DNA
must be long-lived and stable,
Medical Genetics
Problems with integrating
therapeutic DNA into the genome and
the rapidly dividing nature of many
cells prevent gene therapy from
achieving any long-term benefits,
Patients will have to undergo multiple
rounds of gene therapy,
Medical Genetics
B,Immune response
Anytime a foreign object is
introduced into human tissues,the
immune system is designed to attack
the invader,
Medical Genetics
The risk of stimulating the
immune system in a way that
reduces gene therapy effectiveness
is always a potential risk,
Medical Genetics
Furthermore,the immune
system's enhanced response to
invaders it has seen before makes it
difficult for gene therapy to be
repeated in patients,
Medical Genetics
C,Problems with viral vectors
Viruses,while the carrier of
choice in most gene therapy studies,
present a variety of potential
problems to the patient --toxicity,
immune and inflammatory responses,
and gene control and targeting
issues,
Medical Genetics
In addition,there is always the
fear that the viral vector,once inside
the patient,may recover its ability to
cause disease,
Medical Genetics
D,Multigene disorders
Conditions or disorders that arise
from mutations in a single gene are
the best candidates for gene therapy,
Medical Genetics
Unfortunately,some the most
commonly occurring disorders,such
as heart disease,high blood pressure,
Alzheimer's disease,arthritis,and
diabetes,are caused by the
combined effects of variations in
many genes,
Medical Genetics
Multigene or multifactorial
disorders such as these would be
especially difficult to treat effectively
using gene therapy,
Medical Genetics
5,Some recent developments
University of California,Los Angeles,
research team gets genes into the brain
using liposomes coated in a polymer call
polyethylene glycol (PEG),The transfer of
genes into the brain is a significant
achievement because viral vectors are too
big to get across the "blood-brain barrier."
This method has potential for treating
Parkinson's disease,
Medical Genetics
RNA interference or gene
silencing may be a new way to treat
Huntington's,Short pieces of double-
stranded RNA are used by cells to
degrade RNA of a particular
sequence,If a siRNA is designed to
match the RNA copied from a faulty
gene,then the abnormal protein
product of that gene will not be
produced,
Medical Genetics
Medical Genetics
New gene therapy approach
repairs errors in messenger RNA
derived from defective genes,
Technique has potential to treat the
blood disorder thalassaemia,cystic
fibrosis,and some cancers,
Medical Genetics
Gene therapy for treating
children with X-SCID (sever
combined immunodeficiency) or the
"bubble boy" disease is stopped in
France when the treatment causes
leukemia in one of the patients,
Medical Genetics
Researchers at Case Western
Reserve University and Copernicus
Therapeutics are able to create tiny
liposomes 25 nanometers across that
can carry therapeutic DNA through
pores in the nuclear membrane,
Medical Genetics
6,Some of the ethical considerations
? What is normal and what is a
disability or disorder,and who decides?
? Are disabilities diseases? Do they
need to be cured or prevented?
? Does searching for a cure demean the
lives of individuals presently affected
by disabilities?
Medical Genetics
? Who will have access to these
therapies? Who will pay for their use?
? Is somatic gene therapy more or less
ethical than germline gene therapy
(which is done in egg and sperm
cells and prevents the trait from
being passed on to further
generations)?
19 基因治疗
Gene Therapy
Medical Genetics
1,What is gene therapy?
Genes,which are carried on
chromosomes,are the basic physical
and functional units of heredity,
Genes are specific sequences of
bases that encode instructions on
how to make proteins,
Medical Genetics
Although genes get a lot of
attention,it’s the proteins that
perform most life functions and even
make up the majority of cellular
structures,
Medical Genetics
When genes are altered so that
the encoded proteins are unable to
carry out their normal functions,
genetic disorders can result,
Medical Genetics
Gene therapy is a technique for
correcting defective genes
responsible for disease development,
Medical Genetics
Researchers may use one of several approaches
for correcting faulty genes,
1,A normal gene may be inserted into a nonspecific
location within the genome to replace a
nonfunctional gene,This approach is most common,
2,An abnormal gene could be swapped for a normal
gene through homologous recombination,
3,The abnormal gene could be repaired through
selective reverse mutation,which returns the gene
to its normal function,
4,The regulation (the degree to which a gene is turned
on or off) of a particular gene could be altered,
Medical Genetics
2,How does gene therapy work?
In most gene therapy studies,a
"normal" gene is inserted into the
genome to replace an "abnormal,"
disease-causing gene,
Medical Genetics
A carrier molecule called a
vector must be used to deliver the
therapeutic gene to the patient's
target cells,Currently,the most
common vector is a virus that has
been genetically altered to carry
normal human DNA,
Medical Genetics
Viruses have evolved a way of
encapsulating and delivering their
genes to human cells in a pathogenic
manner,
Medical Genetics
Scientists have tried to take
advantage of this capability and
manipulate the virus genome to
remove disease-causing genes and
insert therapeutic genes,
Medical Genetics
Target cells such as the patient's
liver or lung cells are infected with
the viral vector,The vector then
unloads its genetic material
containing the therapeutic human
gene into the target cell,
Medical Genetics
The generation of a functional
protein product from the therapeutic
gene restores the target cell to a
normal state,
Medical Genetics
To reverse disease caused by genetic damage,researchers isolate
normal DNA and package it into a vector,a molecular delivery truck
usually made from a disabled virus,Doctors then infect a target
cell — usually from a tissue affected by the illness,such as liver or
lung cells— with the vector,The vector unloads its DNA cargo,which
then begins producing the missing protein and restores the cell to
normal,
Medical Genetics
A,Some of the different types of
viruses used as gene therapy vectors
(1) Retroviruses
A class of viruses that can create
double-stranded DNA copies of their RNA
genomes,These copies of its genome can
be integrated into the chromosomes of
host cells,Human immunodeficiency virus
(HIV) is a retrovirus,
Medical Genetics
(2)Adenoviruses
A class of viruses with double-stranded DNA
genomes that cause respiratory,intestinal,and
eye infections in humans,The virus that causes
the common cold is an adenovirus,
(3)Adeno-associated viruses
A class of small,single-stranded DNA viruses
that can insert their genetic material at a specific
site on chromosome 19,
(4)Herpes simplex viruses
A class of double-stranded DNA viruses that
infect a particular cell type,neurons,Herpes
simplex virus type 1 is a common human pathogen
that causes cold sores,
Medical Genetics
Medical Genetics
Medical Genetics
Medical Genetics
Besides virus-mediated gene-
delivery systems,there are several
nonviral options for gene delivery,
The simplest method is the direct
introduction of therapeutic DNA into
target cells,This approach is limited
in its application because it can be
used only with certain tissues and
requires large amounts of DNA,
Medical Genetics
Another nonviral approach
involves the creation of an artificial
lipid sphere with an aqueous core,
This liposome,which carries the
therapeutic DNA,is capable of
passing the DNA through the target
cell's membrane,
Medical Genetics
Therapeutic DNA also can get
inside target cells by chemically
linking the DNA to a molecule that
will bind to special cell receptors,
Medical Genetics
Once bound to these receptors,
the therapeutic DNA constructs are
engulfed by the cell membrane and
passed into the interior of the target
cell,This delivery system tends to be
less effective than other options,
Medical Genetics
Researchers also are
experimenting with introducing a
47th artificial human chromosome
into target cells,This chromosome
would exist autonomously alongside
the standard 46 --not affecting their
workings or causing any mutations,
Medical Genetics
It would be a large vector capable of
carrying substantial amounts of genetic
code,and scientists anticipate that,
because of its construction and autonomy,
the body's immune systems would not
attack it,A problem with this potential
method is the difficulty in delivering such
a large molecule to the nucleus of a target
cell,
Medical Genetics
3,The current status of gene therapy research
Current gene therapy is experimental and
has not proven very successful in clinical trials,
Medical Genetics
Little progress has been made
since the first gene therapy clinical
trial began in 1990,In 1999,gene
therapy suffered a major setback
with the death of 18-year-old Jesse
Gelsinger,
Medical Genetics
Jesse was participating in a gene
therapy trial for ornithine
transcarboxylase deficiency (OTCD),
He died from multiple organ failures
4 days after starting the treatment,
His death is believed to have been
triggered by a severe immune
response to the adenovirus carrier,
Medical Genetics
Another major blow came in
January 2003,when the FDA (USA)
placed a temporary halt on all gene
therapy trials using retroviral vectors
in blood stem cells,
Medical Genetics
FDA (USA) took this action after
it learned that a second child treated
in a French gene therapy trial had
developed a leukemia-like condition,
Medical Genetics
Both this child and another who
had developed a similar condition in
August 2002 had been successfully
treated by gene therapy for X-linked
severe combined immunodeficiency
disease (X-SCID),also known as
"bubble baby syndrome."
Medical Genetics
FDA's Biological Response Modifiers
Advisory Committee (BRMAC) met at the
end of February 2003 to discuss possible
measures that could allow a number of
retroviral gene therapy trials for treatment
of life-threatening diseases to proceed
with appropriate safeguards,FDA has yet
to make a decision based on the
discussions and advice of the BRMAC
meeting,
Medical Genetics
4,Factors kept gene therapy effective
A,Short-lived nature of gene therapy
Before gene therapy can become a
permanent cure for any condition,the
therapeutic DNA introduced into target
cells must remain functional and the
cells containing the therapeutic DNA
must be long-lived and stable,
Medical Genetics
Problems with integrating
therapeutic DNA into the genome and
the rapidly dividing nature of many
cells prevent gene therapy from
achieving any long-term benefits,
Patients will have to undergo multiple
rounds of gene therapy,
Medical Genetics
B,Immune response
Anytime a foreign object is
introduced into human tissues,the
immune system is designed to attack
the invader,
Medical Genetics
The risk of stimulating the
immune system in a way that
reduces gene therapy effectiveness
is always a potential risk,
Medical Genetics
Furthermore,the immune
system's enhanced response to
invaders it has seen before makes it
difficult for gene therapy to be
repeated in patients,
Medical Genetics
C,Problems with viral vectors
Viruses,while the carrier of
choice in most gene therapy studies,
present a variety of potential
problems to the patient --toxicity,
immune and inflammatory responses,
and gene control and targeting
issues,
Medical Genetics
In addition,there is always the
fear that the viral vector,once inside
the patient,may recover its ability to
cause disease,
Medical Genetics
D,Multigene disorders
Conditions or disorders that arise
from mutations in a single gene are
the best candidates for gene therapy,
Medical Genetics
Unfortunately,some the most
commonly occurring disorders,such
as heart disease,high blood pressure,
Alzheimer's disease,arthritis,and
diabetes,are caused by the
combined effects of variations in
many genes,
Medical Genetics
Multigene or multifactorial
disorders such as these would be
especially difficult to treat effectively
using gene therapy,
Medical Genetics
5,Some recent developments
University of California,Los Angeles,
research team gets genes into the brain
using liposomes coated in a polymer call
polyethylene glycol (PEG),The transfer of
genes into the brain is a significant
achievement because viral vectors are too
big to get across the "blood-brain barrier."
This method has potential for treating
Parkinson's disease,
Medical Genetics
RNA interference or gene
silencing may be a new way to treat
Huntington's,Short pieces of double-
stranded RNA are used by cells to
degrade RNA of a particular
sequence,If a siRNA is designed to
match the RNA copied from a faulty
gene,then the abnormal protein
product of that gene will not be
produced,
Medical Genetics
Medical Genetics
New gene therapy approach
repairs errors in messenger RNA
derived from defective genes,
Technique has potential to treat the
blood disorder thalassaemia,cystic
fibrosis,and some cancers,
Medical Genetics
Gene therapy for treating
children with X-SCID (sever
combined immunodeficiency) or the
"bubble boy" disease is stopped in
France when the treatment causes
leukemia in one of the patients,
Medical Genetics
Researchers at Case Western
Reserve University and Copernicus
Therapeutics are able to create tiny
liposomes 25 nanometers across that
can carry therapeutic DNA through
pores in the nuclear membrane,
Medical Genetics
6,Some of the ethical considerations
? What is normal and what is a
disability or disorder,and who decides?
? Are disabilities diseases? Do they
need to be cured or prevented?
? Does searching for a cure demean the
lives of individuals presently affected
by disabilities?
Medical Genetics
? Who will have access to these
therapies? Who will pay for their use?
? Is somatic gene therapy more or less
ethical than germline gene therapy
(which is done in egg and sperm
cells and prevents the trait from
being passed on to further
generations)?
