Biologists at UC San Diego have discovered a chemical that offers a
completely new and promising direction for the development of drugs to
treat metabolic disorders such as type 2 diabetes -- a major public
health concern in the United States due to the current obesity epidemic.
(Credit: Image by Peter Allen, UC Santa Barbara
ScienceDaily (July 12, 2012) —
Biologists at UC San Diego have discovered a chemical that offers a
completely new and promising direction for the development of drugs to
treat metabolic disorders such as type 2 diabetes -- a major public
health concern in the United States due to the current obesity epidemic.
Their discovery, detailed in a paper published July 13,2012 in an advance online issue of the journal
Science,
initially came as a surprise because the chemical they isolated does
not directly control glucose production in the liver, but instead
affects the activity of a key protein that regulates the internal
mechanisms of our daily night and day activities, which scientists call
our circadian rhythm or biological clock.
Scientists had long suspected that diabetes and obesity could be
linked to problems in the biological clock. Laboratory mice with altered
biological clocks, for example, often become obese and develop
diabetes. Two years ago, a team headed by Steve Kay, dean of the
Division of Biological Sciences at UC San Diego, discovered the first
biochemical link between the biological clock and diabetes. It found
that a key protein, cryptochrome, that regulates the biological clocks
of plants, insects and mammals also regulates glucose production in the
liver and that altering the levels of this protein could improve the
health of diabetic mice.
Now Kay and his team have discovered a small molecule -- one that can
be easily developed into a drug -- that controls the intricate
molecular cogs or timekeeping mechanisms of cryptochrome in such a
manner that it can repress the production of glucose by the liver. Like
mice and other animals, humans have evolved biochemical mechanisms to
keep a steady supply of glucose flowing to the brain at night, when
we're not eating or otherwise active.
"At the end of the night, our hormones signal that we're in a fasting
state," said Kay. "And during the day, when we're active, our
biological clock shuts down those fasting signals that tell our liver to
make more glucose because that's when we're eating."
Diabetes is caused by an accumulation of glucose in the blood, which
can lead to heart disease, strokes, kidney failure and blindness. In
type 1 diabetes, destruction of insulin producing cells in the pancreas
results in the high blood sugar. In type 2 diabetes, which makes up 90
percent of the cases, gradual resistance to insulin because of obesity
or other problems, leads to high blood sugar.
Kay and his collaborators discovered in 2010 that cryptochrome plays a
critical role in regulating the internal timing of our cyclical eating
patterns, timing our fasting at night with our eating during the day to
maintain a steady supply of glucose in our bloodstream. Other
researchers have recently discovered that cryptochrome also has the
potential to reduce high blood sugar from asthma medication by adjusting
the time of day a patient takes their medication. "We found that if we
increased cryptochrome levels genetically in the liver we could inhibit
the production of glucose by the liver," said Kay.
What he and his team found in their most recent discovery was that a
much smaller molecule, dubbed "KL001" (for the first such compound from
the Kay Lab), can regulate that activity as well. It slowed down the
biological clock by stabilizing the cryptochrome protein -- that is, it
essentially prevented crypotochrome from being sent to the cellular
garbage can, the proteasomes.
The discovery of KL001 was serendipitous, a complete surprise to the
scientists that came about from a parallel effort in Kay's laboratory to
identify molecules that lengthen the biological clock. Two years ago,
Tsuyoshi Hirota, a postdoctoral fellow in Kay's laboratory found a
compound that had the greatest effect ever seen on circadian rhythm, a
chemical the biologists dubbed "longdaysin" because it lengthened the
daily biological clocks of human cells by more than 10 hours.
Continuing his search, Hirota resumed his efforts to find more
chemicals that lengthened or slowed down circadian rhythms, enabling the
scientists to understand more about the intricate chemical and genetic
machinery of the biological clock. He and his colleagues in Kay's lab
did this by screening thousands of compounds from a chemical library
with human cells in individual micro-titer wells in which a luciferase
gene from fireflies is attached to the biological clock machinery,
enabling the scientists to detect a glow whenever the biological clock
is activated. Their molecular fishing expedition came up with a number
of other compounds, one of which was KL001.
"We found other compounds that like longdaysin slowed down the
biological clock," said Kay. "But unlike longdaysin, these compounds did
not inhibit the protein kinases that longdaysin inhibits so we knew
this compound must be working differently. What we needed to know was
what is this compound interacting with? And we were absolutely stunned
when we discovered that what was binding most specifically to our
compound, KL001, was the clock protein cryptochrome that our lab has
worked on in plants, flies and mammals for the last 20 years."
Kay's team turned to biological chemists in Peter Schultz's
laboratory at The Scripps Research Institute to characterize the
compound and understand better chemically how it affected cryptochrome
to lengthen the biological clock.
"Those biochemical studies showed us that KL001 prevents cryptochrome
from being degraded by the proteasome system, which was another big
surprise," said Kay. "It essentially interferes with the signal to send
cryptochrome to the garbage can."
To understand how KL001 worked mechanistically with cryptochrome to
control the biological clock, the team initiated a collaboration with
Frank Doyle and his group at UC Santa Barbara. "They constructed a
beautiful mathematical model of cryptochrome's role in the clock," said
Kay. "That model was essential in allowing us to understand the action
of the compound because the biological clock is very complicated. It's
like opening the back of a Rolex and seeing the hundreds of tiny little
cogs that are tightly integrated."
Based on that mathematical model, the scientists predicted that
adding KL001 to mouse liver cells should stabilize cryptochrome and that
the increased level of cryptochrome would inhibit the production of
enzymes in the liver that stimulate the process of gluconeogenesis --
the generation of glucose during fasting. Experiments done together with
the laboratory of David Brenner, dean of the UC San Diego School of
Medicine and Vice Chancellor for Health Sciences, confirmed that
prediction to be true.
"In mouse liver cells," said Kay, "we showed that KL001 inhibited
gene expression for gluconeogenesis that is induced when exposed to the
hormone glucagon, which promotes glucose production by the liver. It's a
hormone we all produce in fasting states. And our compound, in a dose
dependent way, inhibits hepatic gluconeogenesis, the actual production
of glucose by those liver cells."
Kay said the next step for the research group is to understand how
KL001 and similar molecules that affect cryptochrome function in living
systems, such has laboratory mice. The scientists also plan to probe how
such compounds affect other processes besides the liver that may tie
the biological clock to metabolic diseases. "As with any surprise
discovery," he notes, "this opens the door to more opportunities for
novel therapeutics than we can currently imagine."
Besides Kay, Hirota, Schultz, Doyle and Brenner, other scientists
involved in the discovery were Mariko Sawa, Pagkapol Y. Pongsawakul and
Tim Sonntag of UC San Diego's Division of Biological Sciences; Jae Wook
Lee of TSRI; Peter St. John of UC Santa Barbara; Keiko Iwaisako, Takako
Noguchi and David Welsh of UC San Diego's School of Medicine.
The study was supported by grants from the National Institutes of Health .