Inappropriate growth and survival signaling, which leads to the aberrant growth of cancer cells, is a driving force behind tumors. Much of current cancer research focuses on the kinase enzymes whose mutations are responsible for such disregulated signaling, and many successful molecularly targeted anti-cancer therapeutics are directed at inhibiting kinase activity.
Now, UCLA researchers from the Crump Institute for Molecular Imaging, the Institute for Molecular Medicine, the California NanoSystems Institute, the Jonsson Comprehensive Cancer Center and the department of molecular and medical pharmacology have developed an in vitro method for assessing kinase activity in minute tissue samples from patients. The method involves an integrated microfluidics and imaging platform that can reproducibly measure kinase enzymatic activity from as few as 3,000 cells.
In a paper published Nov. 1 in the journal Cancer Research, the UCLA researchers describe several new technological advances in microfluidics and imaging detection they co-developed to measure kinase activity in small-input samples. The team applied their microfluidic kinase assay to human leukemia patient samples.
"Because the device requires only a very small tissue sample to give results, this method creates new potential for direct kinase experimentation and diagnostics on patient blood, bone marrow and needle biopsy samples," said lead investigator Thomas Graeber, a UCLA professor of molecular and medical pharmacology. "For example, the stem cell properties of leukemia can be directly studied from patient samples."
To improve radio-signal detection, the team used a novel imaging detector, in the form of a solid-state beta camera, which can sensitively detect and spatially resolve radioactive signal directly from a microfluidic chip. The beta camera provides a picture of the activity on the chip, allowing real-time monitoring of the assay performance and outcome. It is highly sensitive and quantitative.
In their first application of the device, the team measured the activity of the mutated kinase responsible for chronic myelogenous leukemia. This mutation is targeted by the clinically successful kinase inhibitor Gleevec.
"We are not aware of other work demonstrating solid-state integrated radioactive imaging from a microfluidic platform," said co-investigator Arion Chatziioannou, a UCLA professor of molecular and medical pharmacology.
The resulting microfluidic in vitro kinase radioassay improves reaction efficiency, compared with standard assays, and can be processed in much less time. This greater efficiency, coupled with the high sensitivity of the beta camera, reduces the amount of sample cell input by two to three orders of magnitude, compared with conventional and 96-well assays. The assay includes a kinase immunocapture step to increase specificity towards the kinase of interest.
"To get the kinase assay to work in a microfluidic environment, we needed to develop new protocols and reagents for efficiently manipulating solid-support kinase capture beads using microfluidic trap-and-release valves," said co-investigator Hsian-Rong Tseng , a UCLA professor of molecular and medical pharmacology.
"Integration of the solid-state beta camera allows researchers to monitor the assay in real time, which proved useful during our protocol development and testing," said Cong Fang, the leading graduate student on the project. "The integrated microfluidic and imaging platform opens new possibilities and makes miniaturization of many common radioactivity-based bioassays to the microfluidic realm possible."
"With the integration of the compact camera, the microfluidic format assay has the potential to be developed into inexpensive bench-top, stand-alone units," said UCLA postdoctoral fellow Nam Vu, who led the imaging development.
"Taken together, the reduced sample input required, the decreased assay time, and the digitally controlled reproducibility of the team's microfluidic kinase radioassay facilitates direct experimentation on clinical samples that are either precious or perishable," said UCLA postdoctoral fellow Yanju Wang, who led the design of the network of microfluidic components that run the assay.
Future experiments will develop reproducible sample collection and measurement conditions for primary patient samples.
Other applications could include profiling of patient and animal model samples for their kinase-inhibitor drug sensitivity, or measurement of kinase activity from stem cells, cancer stem cells and other rare immune cells.
The research team included collaborators from Children's Hospital Los Angeles' division of hematology and oncology and the University of Southern California.
The California NanoSystems Institute at UCLA is an integrated research facility located at UCLA and UC Santa Barbara. Its mission is to foster interdisciplinary collaborations in nanoscience and nanotechnology; to train a new generation of scientists, educators and technology leaders; to generate partnerships with industry; and to contribute to the economic development and the social well-being of California, the United States and the world. The CNSI was established in 2000 with $100 million from the state of California. An additional $850 million of support has come from federal research grants and industry funding. CNSI members are drawn from UCLA's College of Letters and Science, the David Geffen School of Medicine, the School of Dentistry, the School of Public Health and the Henry Samueli School of Engineering and Applied Science. They are engaged in measuring, modifying and manipulating atoms and molecules — the building blocks of our world. Their work is carried out in an integrated laboratory environment. This dynamic research setting has enhanced understanding of phenomena at the nanoscale and promises to produce important discoveries in health, energy, the environment and information technology.
For more news, visit the UCLA Newsroom and follow us on Twitter.
Jennifer Marcus | EurekAlert!
International team discovers novel Alzheimer's disease risk gene among Icelanders
24.10.2016 | Baylor College of Medicine
New bacteria groups, and stunning diversity, discovered underground
24.10.2016 | DOE/Lawrence Berkeley National Laboratory
Researchers from the Institute for Quantum Computing (IQC) at the University of Waterloo led the development of a new extensible wiring technique capable of controlling superconducting quantum bits, representing a significant step towards to the realization of a scalable quantum computer.
"The quantum socket is a wiring method that uses three-dimensional wires based on spring-loaded pins to address individual qubits," said Jeremy Béjanin, a PhD...
In a paper in Scientific Reports, a research team at Worcester Polytechnic Institute describes a novel light-activated phenomenon that could become the basis for applications as diverse as microscopic robotic grippers and more efficient solar cells.
A research team at Worcester Polytechnic Institute (WPI) has developed a revolutionary, light-activated semiconductor nanocomposite material that can be used...
By forcefully embedding two silicon atoms in a diamond matrix, Sandia researchers have demonstrated for the first time on a single chip all the components needed to create a quantum bridge to link quantum computers together.
"People have already built small quantum computers," says Sandia researcher Ryan Camacho. "Maybe the first useful one won't be a single giant quantum computer...
COMPAMED has become the leading international marketplace for suppliers of medical manufacturing. The trade fair, which takes place every November and is co-located to MEDICA in Dusseldorf, has been steadily growing over the past years and shows that medical technology remains a rapidly growing market.
In 2016, the joint pavilion by the IVAM Microtechnology Network, the Product Market “High-tech for Medical Devices”, will be located in Hall 8a again and will...
'Ferroelectric' materials can switch between different states of electrical polarization in response to an external electric field. This flexibility means they show promise for many applications, for example in electronic devices and computer memory. Current ferroelectric materials are highly valued for their thermal and chemical stability and rapid electro-mechanical responses, but creating a material that is scalable down to the tiny sizes needed for technologies like silicon-based semiconductors (Si-based CMOS) has proven challenging.
Now, Hiroshi Funakubo and co-workers at the Tokyo Institute of Technology, in collaboration with researchers across Japan, have conducted experiments to...
14.10.2016 | Event News
14.10.2016 | Event News
12.10.2016 | Event News
24.10.2016 | Power and Electrical Engineering
24.10.2016 | Life Sciences
24.10.2016 | Life Sciences