NEWS / 01.6.

21

Base editing successfully

treats progeria in mice

Credit : Aditya Raguram, Liu lab

Cryo-EM structure of an adenine base editor bound to a guide RNA and a target piece of
DNA.
By Karen Zusi

Correcting the mutation that causes progeria

with base editing leads to strong symptom
reduction and longer lifespan in an animal model
A team led by researchers from the Broad Institute of MIT and
Harvard, the National Institutes of Health, and Vanderbilt
University Medical Center has used base editing, a recently
developed form of gene editing, to rescue disease symptoms and
lifespan in mice that have Hutchinson-Gilford progeria syndrome.

Progeria is a progressive genetic disorder in humans that leads to

death, typically from cardiovascular failure, by the time most
patients reach their mid-teens. The majority of children with this
disorder have a single-letter change in their DNA that causes the
production of a toxic protein.
In this study, mice engineered to carry the progeria mutation, and
demonstrating nearly all features of the human disease, received
a single injection with a base editor programmed to correct the
disease-causing mutation. These mice retained healthy tissues
and lived two and a half times as long as control mice with
progeria that were injected with saline. Indeed, the treated mice
reached an age that corresponds to the beginnings of old age in
healthy mice. 

The study is one of the first examples of using base editing — a

gene editing technique that directly converts an individual DNA
base pair into a different base pair — inside an animal to rescue a
serious genetic disease.

“To our knowledge, this work resulted in the strongest rescue of

the symptoms of progeria by multiple measures,” said co-
corresponding author David Liu, Richard Merkin Professor and
director of the Merkin Institute of Transformative Technologies in
Healthcare at the Broad Institute, professor at Harvard University,
and Howard Hughes Medical Institute investigator. “Five years
ago we were still finishing the development of the very first base
editor. If you had told me then that within five years, a single dose
of a base editor could address progeria in an animal at the DNA,
RNA, protein, vascular pathology, and lifespan levels, I would
have said ‘there’s no way.’ It's a real testament to the dedication
of the team that made this work possible.”

The work paves the way for further preclinical studies developing
a therapy for progeria and provides a blueprint for addressing
other genetic diseases with base editing.

The study appears in Nature with lead author Luke Koblan, a

graduate student in Liu’s lab. Co-senior authors are Francis
Collins, director of the National Institutes of Health and senior
investigator at the National Human Genome Research Institute,
and Jonathan Brown, assistant professor of medicine in the
Division of Cardiovascular Medicine at Vanderbilt University
Medical Center.

Francis Collins, Jonathan Brown, and David Liu describe the

impact of this new study.

“REMARKABLE” RESULTS IN
MICE
The genetic mutation at the root of progeria — a single C-to-T
change in the gene LMNA — was discovered by Collins’s lab in
2003. This single-letter error causes the formation of a toxic
protein called progerin, which damages cells by disrupting the
structure of the nucleus.

To reverse this mutation, the researchers turned to base editing.

The molecular tool, developed in Liu’s lab, uses a programmable
DNA-binding protein such as CRISPR-Cas9 or TALE arrays to
zero in on a specific target gene sequence and another enzyme to
chemically change one nucleotide into another.

The team programmed the base editor ABE 7.10max-VRQR to

target the LMNA gene and convert the mutated T•A base pair
back to the normal C•G pair. When the researchers tested the
base editor in cells taken from children with progeria, 90 percent
of cells were successfully edited, which in turn lowered the levels
of progerin mRNA and protein.
Encouraged by these data, the team began a series of long-term
animal studies, collaborating closely with Jonathan Brown,
Francis Collins, and Mike Erdos, an NIH colleague of Collins who
serves as co-first author of this publication. At the NIH, the
researchers tested the base editor in a mouse model of human
progeria that contained two copies of the mutated
human LMNA gene. The model mice exhibit many hallmarks of
the disease, including progressive deterioration of the arterial
system, skeletal changes, thinning fur, and early death.

Cohorts of mice were given a single injection three days or 14

days after birth with adeno-associated viruses that encoded the
base editor. By six weeks of age, the gene correction was found
in multiple organs at levels ranging from about 10 to 60 percent.

By six months, key organs including the heart of animals injected

with base editor contained much lower levels of progerin than the
saline-injected control mice with progeria. In addition, while the
aorta of control mice showed extensive loss of vascular smooth
muscle cells and high levels of adventitial fibrosis — the
accumulation of fibrotic cells around the aorta — the aorta of mice
injected with the base editor appeared similar to samples from
healthy mice.

“We saw this unexpected, nearly complete, rescue of pathology in

the aorta,” said Liu. “We did a number of follow-up experiments to
make sure we weren't being misled. The aorta samples were
nearly indistinguishable from normal mouse samples, which was
stunning. Cells from the base-edited mice were making the
corrected human lamin A protein instead of progerin.”

The base editing injection also greatly extended the lifespan of the
treated mice and improved their vitality. A healthy mouse lives for
about two years. The treated mice survived to a median of 510
days, which corresponds to the beginning of old age — and is
more than double the length of time that the untreated mice lived.

The dramatic extension in lifespan in the treated mice “was a

really profound result,” Brown said, “because kids with progeria
die early from vascular disease. They suffer from heart attacks
and strokes.”

”When my research lab identified the genetic cause of progeria in

2003, we hoped that someday this might lead to a way to help
these children,” said Collins. “Along the way, we’ve made some
progress with drug therapy, but the potential of actually correcting
the fundamental cause at the DNA level is beyond anything we
could have imagined back then. To see this dramatic response in
our progeria mouse model is one of the most exciting therapeutic
developments I have been part of in 40 years as a physician-
scientist.”

GOING FORWARD
“As a physician-scientist, it’s incredibly exciting to think that an
idea you’ve been working on in the laboratory might actually have
a therapeutic impact,” said Brown. “Ultimately, our goal will be to
try to develop this approach for humans, although there are
remaining questions that we need to address first in these model
systems.”

While the team did not observe significant off-target edits from the
base editor, a number of the longest-living treated mice developed
liver tumors — a known long-term complication when using
adeno-associated viruses to deliver genes into mice.
Additional safety and efficacy studies are underway to further
examine these results and investigate potential ways to mitigate
the risks. The results of the work are being taken forward into
further preclinical studies, with the eventual goal of launching a
clinical trial.

“This work also provides a blueprint for the potential treatment of

other genetic diseases that can be addressed with base editing,”
says Liu. “It amplifies not only our excitement, but also our sense
of responsibility to continue to do careful science while offering
patients the best possible chance to benefit.”

This research was supported in part by the Progeria Research

Foundation, National Institutes of Health (U01AI142756,
UG3AI150551, UG3TR002636, RM1HG009490, R01EB022376,
R35GM118062, Z01HG200305, R01HL146654, R01HL126784),
intramural support from project ZIA HG 200-305-18, and the
Howard Hughes Medical Institute.
Paper(s) cited:
Koblan LW, Erdos MR, et al. In vivo adenine base editing rescues Hutchinson-
Gilford progeria syndrome. Nature. Online January 6, 2021. DOI:
10.1038/s41586-020-03086-7
IN BROAD VIDEO
Base editing successfully treats progeria in mice
 

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