2008 Progress Report – Executive Summary
CCC is dedicated to first use system level control to direct
the behavior of cells toward a desired fate for therapeutic
purposes and then to study the responses of nano-cellular
components in the signaling network using a smart Petri dish
CCC will focus its effort on developing rapid search platforms
for potent drug cocktails. This approach places CCC in a
strategic position to efficiently move along the pathway
toward medicine and to understanding the system response
of nano-cellular molecules under multiple drug stimulations
to explore new combinatory drugs.
Based on the strategy, CCC will realize its vision by setting
the plan as follows:
A-3-a: Rapid Search for Potent Drug Cocktails
Bacteria and viruses evolve rapidly and can become resistant
to a particular drug. If we apply multiple drugs against
infectious agent, it can alleviate the drug resistance problem.
The best known use of drug cocktails has been in the fight
against HIV, the virus that causes AIDS. During two years,
1995-1997, the AIDS death rate dropped 60%. However, not
many diseases have been treated using drug cocktails.
It has been a difficult challenge for researchers to determine
the optimal dose of individual drugs to use in combination.
For instance, a researcher testing 10 different concentrations
of six drugs in every possible arrangement would be faced
with 1 million potential combinations.
The feedback control scheme is to first start with an arbitrary
combination of drug dosages. Quantitative output marker(s)
are chosen and to indicate the outcome of the system, cell
or patent, under drug cocktail stimulation. The output will
be fed into a search algorithm for deciding the next combinatory
drug dosages for possible better system response toward desired
With a feedback control scheme, we have demonstrated the ability
to home in to the most potent drug combinations in tens of
searches instead of performing all one million trials. In
an inhibiting viral infection experiment, the CCC team has
found that total inhibition of the virus occurred at much
lower drug doses than would be necessary if the drugs were
used alone; in fact, the concentrations of the drugs were
about 10 percent of that required when used individually
(Wong et al, PNAS 2008).
A-3-b: Pathway to Medicine
We will use FDA approved drugs for applying drug cocktail
studies to cellular and animal experiments. Once the optimized
drug cocktail is identified, we can move quickly towards
clinical tests. Our approach is a universal methodology that
can be applied to many disease treatments. We have chosen
infection, cancer and regenerative medicine as our targets.
Infectious diseases: The rapid search for potent
combinatory drugs can be applied to diseases infected by
either viruses or bacteria. Our current focus is to study
infection of Herpes Simplex Virus 1 (HSV-1). HSV-1 has strong
clinical relevance due to its high degree of prevalence in
human diseases. Available therapeutics against this pathogen
is only marginally effective and the suppression of disease
reactivation remains elusive. Furthermore, since all available
pharmaceutical agents target one enzyme of the virus, strains
resistant to a particular drug have emerged for various infections.
Cancer: Signal transduction defects originate within
the genome feedback onto the genome to assist in the generation
and/or permissivity of complementary lesions that are required
to drive the carcinogenic process forward. Collection of
cellular components in the signaling pathway network participates
in the process. The process may be interrupted by identifying
key participants and purposeful introduction of offsetting
signals or blockade of the signaling pathway. Drug combinations
can be designed to modify a group of selected pathways that
offset these signals or blockades. We are studying alterations
in key signal transduction pathways, including Jak and Ras
mutations, Pten deficiency, and dysregulated expression of
TCL1. They all function as primary lesions in the development
of a wide array of cancers, including leukemia/lymphoma and
Stem cell studies for regenerative medicine: Among
many important challenges of advancing stem cell for regenerative
medicine, optimizing EB mediated ES cell differentiation
is chosen as our first task. We have demonstrated that hydrophobic
surfaces do not require fetal bovine serum, a potential source
of pathogens, to promote the formation of desired-size EBs.
In addition, our studies hold true for the human EB model.
These two observations were key requirements to obtain high
quality ES. The next step is to develop feedback control
scheme for directing the differentiation of ES amenable for
A-3-c: Smart Petri Dish for Studying Network Responses
of Nano Cellular Components
Exploring and monitoring the behavior of nano-cellular molecules
are important for understanding the overall network response
subjected to multiple external stimulations and for providing
opportunities to develop new drugs. The challenge is to fully
reveal the molecular behavior without missing critical information.
Expression level measurements quantify the numbers of molecules
in cells and provide useful information, but lose critical
information of their interactions in the network of pathways.
We are developing microfluidics-based phosphorylation arrays
which monitor the activities and interactions of cellular
components on a predetermined time course. In addition, we
will be able to directly measure phosphorylation activities
in living cell, once the tunable nano-plasmonic resonator
(TNPR) is developed. Furthermore, the cells under investigation
will be arranged in arrays by opto-electronic tweezers (OET)
which allow us to perform single cell based studies.
Microfluidic circuitry: In the late 1980s, micro-electro-mechanical-system
(MEMS) technologies emerged and enabled us to manufacture
micron scale mechanical devices. A micron size device matches
the size of a cell and facilitates the direct handling of
cells. Fluid handling is necessary in all areas of biotechnology.
Cells are cultured in fluid. Reagents are transported by
fluid. By using microfluidic circuitries, it greatly facilitates
the advancements of medical diagnostic and therapeutic studies.
TNPR: During the past decade, many functional nano-devices
have been developed by the advancement of nanotechnologies.
A nanometer thick sandwich structure, tunable nanoplasmonic
resonator can achieve high level of surface-enhanced Raman
scattering (SERS) signal. With the size of about 50 nm, it
can be inserted into living cell and provide real-time monitoring
of a large number of the cellular functions.
OET based cell processor: The invention of optoelectronic
tweezers (OET) technology allows us to optically-control
cells in a very flexible way. Essentially, we can manipulate
the positions of a large array of cells in a time varying
fashion as we need. Currently, we are integrating OET with
electroporation techniques that have the potential to dramatically
increase the throughput of single-cell-level electroporation.
The integration of these modalities forms a smart Petri dish
which will enable us to perform real time sensing, processing
information and applying stimulations.
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