In collaboration with Uniklinik RWTH Aachen’s neurology department, a Precision²Treat project has amassed a clinical cohort of nearly 300 neuropathic pain patients. These individuals, afflicted with conditions like small fiber neuropathy (SFN) or erythromelalgia (IEM), offer valuable insights into the complexities of chronic pain.

To purchase a high-throughput screening system and fund Precision²Treat, Professor Angelika Lampert from the Institute for Neurophysiology and Professor Ralf Hausmann from the Institute for Clinical Pharmacology received a grant worth €1.7 million from the German Federal Ministry of Education and Research (BMBF).

As reactions to treatment vary widely among people suffering from neuropathic pain, clinical studies have mostly been unsuccessful. Finding a therapy that works often means years of “trial and error” for patients. “Not only is this difficult for the patients, but it’s also frustrating for the relationship between the patients and the doctors treating them,” explains Ralf Hausmann. “So, we would like to know how drugs work on ion channels and whether there are differences in the unmutated or mutated channels.”

Angelika Lampert and Ralf Hausmann’s team work on specific ion channels, including one of the voltage-gated sodium channels. They have seen multiple neuropathic pain patients with a mutation in this ion channel family that alters the way the channel opens and closes. This channel plays an important role in initiating action potential in the nerves.

“We’re looking for mutation changes in the voltage-gated sodium channels Na_v1.7 and Na_v1.8, so we measure their very small electrical current using highly sensitive amplifiers,” explains Angelika Lampert. “We normally do this one cell at a time by manual patch clamp electrophysiology with a tiny little glass pipette and then we check if the current flows in the cell and how the current is characterized.”

It’s a process that works, but it’s also incredibly time consuming. “If you really want to understand the function of a mutation, you need to look at many cells and do many measurements,” says Angelika Lampert.

With Precision²Treat, the team is also expanding their investigation beyond single channel mutations or variants. “We have an enormously high number of channels in our nerve fibers and, ideally, we would like to investigate the genetic variants of the patient by including all these ion channels,” says Angelika Lampert. “It’s almost impossible to do this one channel at a time.”

By taking a patient’s own cells to derive sensory neurons, which carry the complete genetic code with all the variants of that patient, the team will use the Qube 384 high throughput patch clamp system to see if they can modify the cell’s electrophysiological properties by using drugs already on the market. “On the one hand, we are looking for the individual channel when it is expressed in another cell system,” says Ralf Hausmann. “And on the other hand, we have the same channel variant expressed in the so-called natural environment of the channel in the patient cells, which includes all other ion channels and maybe additional mutations in these sodium channels.”

With experience from a previous project funded by the German Federal Ministry for Education and Research (BMBF) that did not include a high throughput automated patch clamp instrument, the team knew this technology would be essential for Precision²Treat. “It also worked in our favor that a funding opportunity came up with BMBF,” highlights Ralf Hausmann. “To get funding for large research equipment is a challenge and a big difference between Industry and Academia.”

“Having a translational project may make it easier to tap into a larger pool of funding,” explains Angelika Lampert. “With a project like Precision²Treat, a wider pool of funding might be applicable because it’s something that the Industry could also be interested in,” continues Angelika Lampert. “Our project has a basic science foundation and a very strong link to clinics with a direct, relatively concrete vision of how this could lead to a treatment for patients.”

In addition to neurologists, the Presicion²Treat research consortium also consists of geneticists, biophysicists, and computer modelers. In contrast to the pharmaceutical industry, the team has detailed phenotyping of all patients along with permission to use their biomaterials for research purposes. “Coordinating our efforts has really helped us make this project a reality – we needed all these specialties under one umbrella,” highlights Ralf Hausmann.

The ambition for Precision²Treat project is to show proof of concept. By using iPSC derived sensory neurons on the Qube to screen licensed drugs, the team hopes to identify compounds that are likely to reduce the excitability of the cells.

“We intend to understand how the biophysical properties could cause the pain symptoms of the patient and then ultimately come to a precision treatment for individualized therapy,” continues Ralf Hausmann.

Now with the Qube 384 in place, the team is adapting iPS cells to the high throughput automated patch clamp system. “This is challenging because it takes about 50 days of culturing for neuronal differentiation and all the sodium channels to be expressed on the cell surface,” explains Angelika Lambert. “And if you do this with neurons, long neurites begin to spread and they must be removed before putting them on the Qube. This can damage the cells, but we have received a lot of help from Sophion, and now we have healthy cells that are working well on our patch clamp system.”

Today the team continues to use manual patch clamp to address specific questions, but the Qube is now giving them more possibilities and enabling them to do the kind of high throughput drug screening that would have been impossible with their previous setup. “Before, we would see a variant of a patient or a certain mutation in the sodium channel that is localized at the voltage sensor, and then we would analyze this specific property with manual patch clamp,” explains Ralf Hausmann. “But now we can run a pool of protocols analyzing most of the typical channel properties.”

Supporting an optimized research workflow, the Qube enables the team to analyze hundreds of compounds and have the results in just a day or two. “It’s much faster – you look at the data and decide what your next step will be,” says Ralf Hausmann.

The Aachen team tested the different automated patch clamp systems available, keeping the highly specific needs of the Precision2Treat project top of mind. “The sensory neurons are very large cells and therefore have special requirements such as full Rs compensation for a precise biophysical analysis, and this is available in the Qube,” adds Angelika Lampert.

The team was also pleased with the installation and the on-boarding provided by Sophion. “A team of field engineers and application scientists came to Aachen to set up the Qube,” says Ralf Hausmann. “We got an introduction to the system as well as an understanding of how to troubleshoot and get assistance from their subject matter experts. You get the right answer from the right specialist for any question. Plus, they have extensive experience with different cell types and channels, so sometimes you can speed up development just by asking the Sophion team.”

The experience has also been collaborative, with the Aachen team excited to share ideas for features and future developments with the Sophion team. “The experience has been highly professional and very friendly,” says Angelika Lampert. “When we have new ideas in the project, the Sophion team is always interested in seeing how we can work together to improve the system even more. So, this is a fun aspect of our work together.”

Founded at Uniklinik RWTH Aachen in 2023, the Scientific Centre for Neuropathic Pain Aachen (SCNa) brings together experts in the field.