InSphero’s breakthrough in generating 3D cell models of human organs at scale for use in toxicity screens attracted potential customers to invest in proving the technology. Now all the top ten pharma companies are users
Fail early, fail fast may not seem a very inspiring mantra. But that is exactly what pharmaceutical companies must do to avoid wasting money on compounds that flop in expensive clinical trials because toxicities come to light that were not evident in pre-clinical testing.
Of course, no company would advance a product that was likely to cause toxic side effects. But tools for detecting toxicity in pre-clinical laboratory screening are inadequate.
In particular, the industry currently relies heavily on 2D cellular screens, in which cells are spread one layer deep across a substrate. This is a poor substitute for the complexities of a human organ like the liver, as evidenced by the fact that liver toxicity is one of the most common reasons for drugs to fail when tested in humans.
Conventional pre-clinical toxicology testing fails to predict human liver toxicity in approximately 50 per cent of cases.
InSphero’s leap forward has been in translating microfluidics research carried out at ETH Zurich and the University of Zurich, into a robust technique for generating 3D models of human organs at a scale, speed and cost that makes them suitable for high throughput screening.
Self-assembly
Rather than a 2D monolayer in which individual cells interact with the substrate, InSphero’s surface-free hanging drop technology prompts cells to interact with each other. Suspended in tiny drops of culture media, cells behave as they are programmed to do in the three-dimensional human body, signalling to other cells and self-assembling into 3D tissue structures that better mimic human organs, creating more physiologically relevant models.
Although hanging drop culture has been in use for many years, it was a painstaking and slow manual process. “We have taken a very simple principle and made it work at scale,” said Jan Lichtenberg, co-founder and CEO of InSphero. “It takes 20 seconds to seed an industry-standard 384-well plate, and two-to-four days to incubate the cells while they self-assemble,” he told Science|Business.
InSphero won the 2014 Science|Business Academic Enterprise Award for Life Sciences announced at the Technischen Universität Berlin earlier this month.
Repeat dosing
Using the best 2D culture methods, liver cells lose their specificity within two to four days. In contrast, InSphero’s 3D InSight liver model retains its integrity for 60 days. “With 2D cell assays you can only really test for acute toxicity. But when a product is pulled from the market because of liver toxicity, it’s not because it was obvious at two days, but because the toxicity took two weeks to show up,” Lichtenberg said.
The 60-day life span of the 3D InSight liver model makes it possible to do repeat dose testing that mimics how humans take drugs. It also more closely mimics the high rate of metabolic turnover that occurs in the liver. This is significant because a toxic effect may not be caused by a compound per se, but by one of the metabolites to which it is broken down in the liver.
In one demonstration test, InSphero detected liver toxicity caused by Pfizer’s antibiotic Trovan (trovafloxacin) in a screen costing $5,000. Pfizer became one of InSphero’s biggest customers, and no wonder: Trovan made it all the way through pre-clinical and clinical testing before liver toxicity led to it being withdrawn after a year on the market, at an estimated overall cost of $8.5 billion.
InSphero has developed sand grain-size 3D models of other organs including the heart, skin and pancreas. It also markets the underlying technology platform, enabling customers to develop their own models.
A bigger vision
Lichtenberg was a post doc at ETH Zurich when he did the original research with InSphero’s chief scientific officer, Jens Kelm, then a PhD student. “The research was interesting but I left soon after because I wanted to get commercial experience,” Lichtenberg said.
His motivation came from having formed and worked with an earlier start-up, Advanced Micromachining Tools (AMT), from 1999 – 2004. AMT was based on technology Lichtenberg developed as an undergraduate at the University of Bremen. “The company is still there and I really enjoyed doing it and was keen to repeat the experience. However, for the next company, I wanted a bigger vision and saw the need to work in industry first,” said Lichtenberg.
Kelm continued to work on the hanging drop technology and by 2008 had succeeded in developing a fine-tuned, automated process, providing the spur to start thinking about commercialisation with the third co-founder, Wolfgang Moritz, InSphero’s head of R&D.
In 2009 and 2010, InSphero won business plan awards totalling €220,000, enabling it to in-license technology from ETH Zurich and from the University of Zurich, and become independent of its academic roots.
In parallel, the co-founders contacted potential customers, including Novartis, to assess their interest. “Novartis was impressed by the technology and agreed to invest in a collaboration if we agreed to work on the company full-time,” Lichtenberg said. Other potential customers, including the cosmetics firm L’Oreal and the Swiss medical biotechnology company Actelion Pharmaceuticals, made similar stipulations.
Do quit the day job
“We saw no industrial customers would work with us unless we quit our jobs. Everyone said we would be crazy to do so because this was 2009 and the beginning of the financial downturn,” Lichtenberg recalled. But having the promise of backing from potential users gave the founders confidence to resign and by 1 January 2010, the trio was working full-time at InSphero.
The support from industry also provided the validation needed to raise a Series A venture capital and business angel round of €1.5 million in 2010. In 2012/13 InSphero completed a follow-on financing in equity and credit, to support expansion of the business in Europe and the US. It has also been awarded three European Union and two Swiss R&D grants totalling €1.2 million.
While the start of a global financial crisis may seem a mad moment to found a business, the timing was auspicious for several reasons. The pharmaceutical industry was becoming increasingly concerned about declining R&D productivity and putting increasing effort into confronting the problem. In parallel, the FDA and the European Medicines Agency were looking to reduce the risks for patients and calling for the development of more biologically relevant cellular models.
This impetus to increase the predictive power of cell-based screening was heightened by pressure to reduce animal testing. The EU banned all animal testing of cosmetics in member states in 2009, following this up in 2013 with a complete ban on the sale of any cosmetics tested in animals, regardless of where in the world the tests were conducted.
Patent expiries
At the same time, the potential to generate better cell-based models was enhanced by advances in stem cell technology and the invention in 2006 of induced pluripotent stem cells, increasing the supply and diversity of cell types that are available as the basis of models.
“In 2003 – 04 when the initial work on our technology was carried out, the pharma industry was still in the era of blockbuster drugs,” Lichtenberg noted. “They had great products and wouldn’t have been interested.” But subsequent patent expiries were not matched by new launches, forcing the industry to focus on ways to improve productivity.
Timing apart, the other important element in InSphero’s success has been its knowledge of the industry’s requirements. “We made a lot of mistakes, but the one thing we did really well was to start selling products at the earliest stages of development. We would go and talk to customers, explain the capabilities and see if they fitted their needs and workflow,” Lichtenberg said.
Proof of the value of the technology comes from InSphero’s roster of customers. “I’m particularly proud of the industry pick-up. All of the top ten pharma companies are using our technology, or evaluating it at a practical level,” Lichtenberg concluded.
Of course, no company would advance a product that was likely to cause toxic side effects. But tools for detecting toxicity in pre-clinical laboratory screening are inadequate.
In particular, the industry currently relies heavily on 2D cellular screens, in which cells are spread one layer deep across a substrate. This is a poor substitute for the complexities of a human organ like the liver, as evidenced by the fact that liver toxicity is one of the most common reasons for drugs to fail when tested in humans.
Conventional pre-clinical toxicology testing fails to predict human liver toxicity in approximately 50 per cent of cases.
InSphero’s leap forward has been in translating microfluidics research carried out at ETH Zurich and the University of Zurich, into a robust technique for generating 3D models of human organs at a scale, speed and cost that makes them suitable for high throughput screening.
Self-assembly
Rather than a 2D monolayer in which individual cells interact with the substrate, InSphero’s surface-free hanging drop technology prompts cells to interact with each other. Suspended in tiny drops of culture media, cells behave as they are programmed to do in the three-dimensional human body, signalling to other cells and self-assembling into 3D tissue structures that better mimic human organs, creating more physiologically relevant models.
Although hanging drop culture has been in use for many years, it was a painstaking and slow manual process. “We have taken a very simple principle and made it work at scale,” said Jan Lichtenberg, co-founder and CEO of InSphero. “It takes 20 seconds to seed an industry-standard 384-well plate, and two-to-four days to incubate the cells while they self-assemble,” he told Science|Business.
InSphero won the 2014 Science|Business Academic Enterprise Award for Life Sciences announced at the Technischen Universität Berlin earlier this month.
Repeat dosing
Using the best 2D culture methods, liver cells lose their specificity within two to four days. In contrast, InSphero’s 3D InSight liver model retains its integrity for 60 days. “With 2D cell assays you can only really test for acute toxicity. But when a product is pulled from the market because of liver toxicity, it’s not because it was obvious at two days, but because the toxicity took two weeks to show up,” Lichtenberg said.
The 60-day life span of the 3D InSight liver model makes it possible to do repeat dose testing that mimics how humans take drugs. It also more closely mimics the high rate of metabolic turnover that occurs in the liver. This is significant because a toxic effect may not be caused by a compound per se, but by one of the metabolites to which it is broken down in the liver.
In one demonstration test, InSphero detected liver toxicity caused by Pfizer’s antibiotic Trovan (trovafloxacin) in a screen costing $5,000. Pfizer became one of InSphero’s biggest customers, and no wonder: Trovan made it all the way through pre-clinical and clinical testing before liver toxicity led to it being withdrawn after a year on the market, at an estimated overall cost of $8.5 billion.
InSphero has developed sand grain-size 3D models of other organs including the heart, skin and pancreas. It also markets the underlying technology platform, enabling customers to develop their own models.
A bigger vision
Lichtenberg was a post doc at ETH Zurich when he did the original research with InSphero’s chief scientific officer, Jens Kelm, then a PhD student. “The research was interesting but I left soon after because I wanted to get commercial experience,” Lichtenberg said.
His motivation came from having formed and worked with an earlier start-up, Advanced Micromachining Tools (AMT), from 1999 – 2004. AMT was based on technology Lichtenberg developed as an undergraduate at the University of Bremen. “The company is still there and I really enjoyed doing it and was keen to repeat the experience. However, for the next company, I wanted a bigger vision and saw the need to work in industry first,” said Lichtenberg.
Kelm continued to work on the hanging drop technology and by 2008 had succeeded in developing a fine-tuned, automated process, providing the spur to start thinking about commercialisation with the third co-founder, Wolfgang Moritz, InSphero’s head of R&D.
In 2009 and 2010, InSphero won business plan awards totalling €220,000, enabling it to in-license technology from ETH Zurich and from the University of Zurich, and become independent of its academic roots.
In parallel, the co-founders contacted potential customers, including Novartis, to assess their interest. “Novartis was impressed by the technology and agreed to invest in a collaboration if we agreed to work on the company full-time,” Lichtenberg said. Other potential customers, including the cosmetics firm L’Oreal and the Swiss medical biotechnology company Actelion Pharmaceuticals, made similar stipulations.
Do quit the day job
“We saw no industrial customers would work with us unless we quit our jobs. Everyone said we would be crazy to do so because this was 2009 and the beginning of the financial downturn,” Lichtenberg recalled. But having the promise of backing from potential users gave the founders confidence to resign and by 1 January 2010, the trio was working full-time at InSphero.
The support from industry also provided the validation needed to raise a Series A venture capital and business angel round of €1.5 million in 2010. In 2012/13 InSphero completed a follow-on financing in equity and credit, to support expansion of the business in Europe and the US. It has also been awarded three European Union and two Swiss R&D grants totalling €1.2 million.
While the start of a global financial crisis may seem a mad moment to found a business, the timing was auspicious for several reasons. The pharmaceutical industry was becoming increasingly concerned about declining R&D productivity and putting increasing effort into confronting the problem. In parallel, the FDA and the European Medicines Agency were looking to reduce the risks for patients and calling for the development of more biologically relevant cellular models.
This impetus to increase the predictive power of cell-based screening was heightened by pressure to reduce animal testing. The EU banned all animal testing of cosmetics in member states in 2009, following this up in 2013 with a complete ban on the sale of any cosmetics tested in animals, regardless of where in the world the tests were conducted.
Patent expiries
At the same time, the potential to generate better cell-based models was enhanced by advances in stem cell technology and the invention in 2006 of induced pluripotent stem cells, increasing the supply and diversity of cell types that are available as the basis of models.
“In 2003 – 04 when the initial work on our technology was carried out, the pharma industry was still in the era of blockbuster drugs,” Lichtenberg noted. “They had great products and wouldn’t have been interested.” But subsequent patent expiries were not matched by new launches, forcing the industry to focus on ways to improve productivity.
Timing apart, the other important element in InSphero’s success has been its knowledge of the industry’s requirements. “We made a lot of mistakes, but the one thing we did really well was to start selling products at the earliest stages of development. We would go and talk to customers, explain the capabilities and see if they fitted their needs and workflow,” Lichtenberg said.
Proof of the value of the technology comes from InSphero’s roster of customers. “I’m particularly proud of the industry pick-up. All of the top ten pharma companies are using our technology, or evaluating it at a practical level,” Lichtenberg concluded.
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