Tag Archives: microfluidic systems

Human-on-a-chip predicts in vivo results based on in vitro model … for the first time

If successful the hope is that ‘human-on-a-chip’ will replace most, if not all, animal testing. This July 3, 2019 Hesperos news release (also on EurekAlert) suggests scientists are making serious gains in the drive to replace animal testing (Note: For anyone having difficulty with the terms, pharmacokinetics and pharmacodynamics, there are definitions towards the end of this posting, which may prove helpful),

Hesperos Inc., pioneers* of the “human-on-a-chip” in vitro system has announced the use of its innovative multi-organ model to successfully measure the concentration and metabolism of two known cardiotoxic small molecules over time, to accurately describe the drug behavior and toxic effects in vivo. The findings further support the potential of body-on-a-chip systems to transform the drug discovery process.

In a study published in Nature Scientific Reports, in collaboration with AstraZeneca, Hesperos described how they used a pumpless heart model and a heart:liver system to evaluate the temporal pharmacokinetic/pharmacodynamic (PKPD) relationship for terfenadine, an antihistamine that was banned due to toxic cardiac effects, as well as determine its mechanism of toxicity.

The study found there was a time-dependent, drug-induced response in the heart model. Further experiments were conducted, adding a metabolically competent liver module to the Hesperos Human-on-a-Chip® system to observe what happened when terfenadine was converted to fexofenadine. By doing so, the researchers were able to determine the driver of the pharmacodynamic (PD) effect and develop a mathematical model to predict the effect of terfenadine in preclinical species. This is the first time an in vitro human-on-a-chip system has been shown to predict in vivo outcomes, which could be used to predict clinical trial outcomes in the future.

“The ability to examine PKPD relationships in vitro would enable us to understand compound behavior prior to in vivo testing, offering significant cost and time savings,” said Dr. Shuler, President and CEO, Hesperos, Inc and Professor Emeritus, Cornell University. “We are excited about the potential of this technology to help us ensure that potential new drug candidates have a higher probability of success during the clinical trial process.”

Understanding the inter-relationship between pharmacokinetics (PK), the drug’s time course for absorption, distribution, metabolism and excretion, and PD, the biological effect of a drug, is crucial in drug discovery and development. Scientists have learned that the maximum drug effect is not always driven by the peak drug concentration. In some cases, time is a critical factor influencing drug effect, but often this concentration-effect-time relationship only comes to light during the advanced stages of the preclinical program. In addition, often the data cannot be reliably extrapolated to humans.

“It is costly and time consuming to discover that potential drug candidates may have poor therapeutic qualities preventing their onward progression,” said James Hickman, Chief Scientist at Hesperos and Professor at the University of Central Florida. “Being able to define this during early drug discovery will be a valuable contribution to the optimization of potential new drug candidates.”

As demonstrated with the terfenadine experiment, the PKPD modelling approach was critical for understanding both the flux of compound between compartments as well as the resulting PD response in the context of dynamic exposure profiles of both parent and metabolite, as indicated by Dr. Shuler.

In order to test the viability of their system in a real-world drug discovery setting, the Hesperos team collaborated with scientists at AstraZeneca, to test one of their failed small molecules, known to have a CV [cardiovscular?] risk.

One of the main measurements used to assess the electrical properties of the heart is the QT interval, which approximates the time taken from when the cardiac ventricles start to contract to when they finish relaxing. Prolongation of the QT interval on the electrocardiogram can lead to a fatal arrhythmia known as Torsade de Pointes. Consequently, it is a mandatory requirement prior to first-in-human administration of potential new drug candidates that their ability to inhibit the hERG channel (a biomarker for QT prolongation) is investigated.

In the case of the AstraZeneca molecule, the molecule was assessed for hERG inhibition early on, and it was concluded to have a low potential to cause in vivo QT prolongation up to 100 μM. In later pre-clinical testing, the QT interval increased by 22% at a concentration of just 3 μM. Subsequent investigations found that a major metabolite was responsible. Hesperos was able to detect a clear PD effect at concentrations above 3 μM and worked to determine the mechanism of toxicity of the molecule.

The ability of these systems to assess cardiac function non-invasively in the presence of both parent molecule and metabolite over time, using multiplexed and repeat drug dosing regimes, provides an opportunity to run long-term studies for chronic administration of drugs to study their potential toxic effects.

Hesperos, Inc. is the first company spun out from the Tissue Chip Program at NCATS (National Center for Advancing Translational Sciences), which was established in 2011 to address the long timelines, steep costs and high failure rates associated with the drug development process. Hesperos currently is funded through NCATS’ Small Business Innovation Research program to undertake these studies and make tissue chips technology available as a service based company.

“The application of tissue chip technology in drug testing can lead to advances in predicting the potential effects of candidate medicines in people,” said Danilo Tagle, Ph.D., associate director for special initiatives at NCATS.


About Hesperos
Hesperos, Inc. is a leader in efforts to characterize an individual’s biology with human-on-a-chip microfluidic systems. Founders Michael L. Shuler and James J. Hickman have been at the forefront of every major scientific discovery in this realm, from individual organ-on-a-chip constructs to fully functional, interconnected multi-organ systems. With a mission to revolutionize toxicology testing as well as efficacy evaluation for drug discovery, the company has created pumpless platforms with serum-free cellular mediums that allow multi-organ system communication and integrated computational PKPD modeling of live physiological responses utilizing functional readouts from neurons, cardiac, muscle, barrier tissues and neuromuscular junctions as well as responses from liver, pancreas and barrier tissues. Created from human stem cells, the fully human systems are the first in vitro solutions that accurately utilize in vitro systems to predict in vivo functions without the use of animal models, as featured in Science. More information is available at http://www.

Years ago I went to a congress focused on alternatives to animal testing (August 22, 2014 posting) and saw a video of heart cells in a petri dish (in vitro) beating in a heartlike rhythm. It was something like this,

Published on Oct 17, 2010 https://www.youtube.com/watch?v=BqzW9Jq-OVA

I found it amazing as did the scientist who drew my attention to it. After, it’s just a collection of heart cells. How do they start beating and keep time with each other?

Getting back to the latest research, here’s a link and a citation for the paper,

On the potential of in vitro organ-chip models to define temporal pharmacokinetic-pharmacodynamic relationships by Christopher W. McAleer, Amy Pointon, Christopher J. Long, Rocky L. Brighton, Benjamin D. Wilkin, L. Richard Bridges, Narasimham Narasimhan Sriram, Kristin Fabre, Robin McDougall, Victorine P. Muse, Jerome T. Mettetal, Abhishek Srivastava, Dominic Williams, Mark T. Schnepper, Jeff L. Roles, Michael L. Shuler, James J. Hickman & Lorna Ewart. Scientific Reports volume 9, Article number: 9619 (2019) DOI: https://doi.org/10.1038/s41598-019-45656-4 Published: 03 July 2019

This paper is open access.

I happened to look at the paper and found good definitions of pharmacokinetics and pharmacodynamics. I know it’s not for everyone but if you’ve ever been curious about the difference (from the Introduction of On the potential of in vitro organ-chip models to define temporal pharmacokinetic-pharmacodynamic relationships),

Integrative pharmacology is a discipline that builds an understanding of the inter-relationship between pharmacokinetics (PK), the drug’s time course for absorption, distribution, metabolism and excretion and pharmacodynamics (PD), the biological effect of a drug. In drug discovery, this multi-variate approach guides medicinal chemists to modify structural properties of a drug molecule to improve its chance of becoming a medicine in a process known as “lead optimization”.

*More than one person and more than one company and more than one country claims pioneer status where ‘human-on-a-chip’ is concerned.

Controlling water with ‘stick-slip surfaces’

Controlling water could lead to better designed microfluidic devices such as ‘lab-on-a-chip’. A July 27, 2015 news item on Nanowerk announces a new technique for controlling water,

Coating the inside of glass microtubes with a polymer hydrogel material dramatically alters the way capillary forces draw water into the tiny structures, researchers have found. The discovery could provide a new way to control microfluidic systems, including popular lab-on-a-chip devices.

Capillary action draws water and other liquids into confined spaces such as tubes, straws, wicks and paper towels, and the flow rate can be predicted using a simple hydrodynamic analysis. But a chance observation by researchers at the Georgia Institute of Technology [US] will cause a recalculation of those predictions for conditions in which hydrogel films line the tubes carrying water-based liquids.

“Rather than moving according to conventional expectations, water-based liquids slip to a new location in the tube, get stuck, then slip again – and the process repeats over and over again,” explained Andrei Fedorov, a professor in the George W. Woodruff School of Mechanical Engineering at Georgia Tech. “Instead of filling the tube with a rate of liquid penetration that slows with time, the water propagates at a nearly constant speed into the hydrogel-coated capillary. This was very different from what we had expected.”

A July 27, 2015 Georgia Institute of Technology (Georgia Tech) news release (also on EurekAlert) by John Toon, which originated the news item, describes the work in more detail,

When the opening of a thin glass tube is exposed to a droplet of water, the liquid begins to flow into the tube, pulled by a combination of surface tension in the liquid and adhesion between the liquid and the walls of the tube. Leading the way is a meniscus, a curved surface of the water at the leading edge of the water column. An ordinary borosilicate glass tube fills by capillary action at a gradually decreasing rate with the speed of meniscus propagation slowing as a square root of time.

But when the inside of a tube is coated with a very thin layer of poly(N-isopropylacrylamide), a so-called “smart” polymer (PNIPAM), everything changes. Water entering a tube coated on the inside with a dry hydrogel film must first wet the film and allow it to swell before it can proceed farther into the tube. The wetting and swelling take place not continuously, but with discrete steps in which the water meniscus first sticks and its motion remains arrested while the polymer layer locally deforms. The meniscus then rapidly slides for a short distance before the process repeats. This “stick-slip” process forces the water to move into the tube in a step-by-step motion.

The flow rate measured by the researchers in the coated tube is three orders of magnitude less than the flow rate in an uncoated tube. A linear equation describes the time dependence of the filling process instead of a classical quadratic equation which describes filling of an uncoated tube.

“Instead of filling the capillary in a hundredth of a second, it might take tens of seconds to fill the same capillary,” said Fedorov. “Though there is some swelling of the hydrogel upon contact with water, the change in the tube diameter is negligible due to the small thickness of the hydrogel layer. This is why we were so surprised when we first observed such a dramatic slow-down of the filing process in our experiments.”

The researchers – who included graduate students James Silva, Drew Loney and Ren Geryak and senior research engineer Peter Kottke – tried the experiment again using glycerol, a liquid that is not absorbed by the hydrogel. With glycerol, the capillary action proceeded through the hydrogel-coated microtube as with an uncoated tube in agreement with conventional theory. After using high-resolution optical visualization to study the meniscus propagation while the polymer swelled, the researchers realized they could put this previously-unknown behavior to good use.

Water absorption by the hydrogels occurs only when the materials remain below a specific transition temperature. When heated above that temperature, the materials no longer absorb water, eliminating the “stick-slip” phenomenon in the microtubes and allowing them to behave like ordinary tubes.

This ability to turn the stick-slip behavior on and off with temperature could provide a new way to control the flow of water-based liquid in microfluidic devices, including labs-on-a-chip. The transition temperature can be controlled by varying the chemical composition of the hydrogel.

“By locally heating or cooling the polymer inside a microfluidic chamber, you can either speed up the filling process or slow it down,” Fedorov said. “The time it takes for the liquid to travel the same distance can be varied up to three orders of magnitude. That would allow precise control of fluid flow on demand using external stimuli to change polymer film behavior.”

The heating or cooling could be done locally with lasers, tiny heaters, or thermoelectric devices placed at specific locations in the microfluidic devices.

That could allow precise timing of reactions in microfluidic devices by controlling the rate of reactant delivery and product removal, or allow a sequence of fast and slow reactions to occur. Another important application could be controlled drug release in which the desired rate of molecule delivery could be dynamically tuned over time to achieve the optimal therapeutic outcome.

In future work, Fedorov and his team hope to learn more about the physics of the hydrogel-modified capillaries and study capillary flow using partially-transparent microtubes. They also want to explore other “smart” polymers which change the flow rate in response to different stimuli, including the changing pH of the liquid, exposure to electromagnetic radiation, or the induction of mechanical stress – all of which can change the properties of a particular hydrogel designed to be responsive to those triggers.

“These experimental and theoretical results provide a new conceptual framework for liquid motion confined by soft, dynamically evolving polymer interfaces in which the system creates an energy barrier to further motion through elasto-capillary deformation, and then lowers the barrier through diffusive softening,” the paper’s authors wrote. “This insight has implications for optimal design of microfluidic and lab-on-a-chip devices based on stimuli-responsive smart polymers.”

In addition to those already mentioned, the research team included Professor Vladimir Tsukruk from the Georgia Tech School of Materials Science and Engineering and Rajesh Naik, Biotechnology Lead and Tech Advisor of the Nanostructured and Biological Materials Branch of the Air Force Research Laboratory (AFRL).

Here’s a link to and a citation for the paper,

Stick–slip water penetration into capillaries coated with swelling hydrogel by J. E. Silva, R. Geryak, D. A. Loney, P. A. Kottke, R. R. Naik, V. V. Tsukruk, and A. G. Fedorov. Soft Matter, 2015,11, 5933-5939 DOI: 10.1039/C5SM00660K First published online 23 Jun 2015

This paper is behind a paywall.