Periodic Reporting for period 2 - SiMulTox (Simultaneous Multiparametric MEA based platform for in-vitro chronic cardiotoxicity assessment with live-cell fluorescence imaging and electrophysiology.)
Reporting period: 2023-04-01 to 2024-06-30
Drug related adverse effects (AEs) that affect cardiac function are the main reasons for drug attrition since these can result in proarrhythmia and life-threatening serious adverse events. Being able to predict these AEs early in drug development could speed up the time it takes to bring a drug to market saving vast amounts of resources. To be approved in the United States, FDA requires toxicity tests for each drug on one rodent species and one nonrodent species. Therefore, biotech and pharma companies use tens of thousands of animals for such tests each year. However, more than 9/10 drugs that enter human clinical trials fail because they are unsafe or ineffective, showing that animal experiments are not conclusive.
Due to this reason, FDA has an impressive track record of pushing the translation of new technologies like in vitro methods to improve safety predictions, and their Modernization Act is the most recent example of this.
In the context of improving in vitro methods for accurate cardiotoxicity prediction, Foresee Biosystems (FB) developed IntraCell, a novel product for the long-term analysis of Action Potentials (APs) from cardiac cells over many weeks, providing information over prolonged treatments with experimental drugs. Going beyond IntraCell, SiMulTox aims at developing a novel tool to simultaneously assess in vitro chronic functional and structural cardiotoxicity on the same biological sample. This new product will enable the multiparametric assessment of drug toxic effects very early in the drug development process, thereby reducing risk of unsuccessful clinical stages. It is very well-known that many chemicals can affect not only the electrical activity of the cardiac tissue, but also other biological features, such as, protein expression and localization, metabolism and subcellular organization. To address the need for more reliable platforms for pre-clinical assessment and safety pharmacology, SiMulTox aims at coupling live-cell fluorescence imaging with FB advanced electrophysiological measurement capabilities. This feature is pivotal in the emerging and continuosly growing market of multiplexed platforms for toxicology, because every medication intended for use in humans must be tested before the earliest clinical trial phase to reduce the risk of future heart damage.
Due to this reason, FDA has an impressive track record of pushing the translation of new technologies like in vitro methods to improve safety predictions, and their Modernization Act is the most recent example of this.
In the context of improving in vitro methods for accurate cardiotoxicity prediction, Foresee Biosystems (FB) developed IntraCell, a novel product for the long-term analysis of Action Potentials (APs) from cardiac cells over many weeks, providing information over prolonged treatments with experimental drugs. Going beyond IntraCell, SiMulTox aims at developing a novel tool to simultaneously assess in vitro chronic functional and structural cardiotoxicity on the same biological sample. This new product will enable the multiparametric assessment of drug toxic effects very early in the drug development process, thereby reducing risk of unsuccessful clinical stages. It is very well-known that many chemicals can affect not only the electrical activity of the cardiac tissue, but also other biological features, such as, protein expression and localization, metabolism and subcellular organization. To address the need for more reliable platforms for pre-clinical assessment and safety pharmacology, SiMulTox aims at coupling live-cell fluorescence imaging with FB advanced electrophysiological measurement capabilities. This feature is pivotal in the emerging and continuosly growing market of multiplexed platforms for toxicology, because every medication intended for use in humans must be tested before the earliest clinical trial phase to reduce the risk of future heart damage.
The first year of the project focused both on the technical and the business development of a new product that will approach the emerging market of multiplexed systems for in vitro evaluation of toxicological outcomes of xenobiotic compounds. The technical includes three different technological segments: optical design, mechanical design, software development. The main achievement of year 1 has been the realization of the core opto-mechanical apparatus for parallel laser stimulation and fluorescence detection from cardiac cells. Great efforts have been dedicated to the study and selection of all the optical components followed by a proper mechanical design of the optical support cage.
A set of LED sources with corresponding excitation filters have been included within the optical path for sample fluorescence excitation under 3 well defined wavelengths. A highly sensitive fluorescence detector has been aligned on the output light path in order to collect fluorescence signals. This structure has been conceived to works in parallel with the proprietary laser-based technology developed by Foresee Biosystems. Though a fully automated 3-steps procedure (laser focus, laser auto-alignment and laser scan over the MEA electrodes), cardiac cells are simultaneously analysed both functionally and structurally. Two different objective lenses are exploited to focus laser and RGB light over the sample to provide high quality AP recordings (useful for analysis of defects in ion channels functionality) and sensitive fluorescence detection (useful for structural changes of cells).
In parallel, we worked on a novel enclosure case, including all the necessary electronic and mechanical support parts, focusing on the exploitation of material and fabrication approaches suitable for biomedical labs equipment.
A software graphical interface is under development. It allows for the parallel management of both laser scan procedure and fluorescence data acquisition from the detectors: a scientific CMOS camera equipped with sensitive sensors for fluorescence imaging and an industrial camera suitable for the Infrared imaging of the sample.
The software interface will include sub-libraries for filters and objective movements during the normal use, allowing an easy switch from the functional to the structural analysis of the samples.
In addition, a deep communication activity has been carried out to speed up SiMulTox outreach to the stakeholders. The main activity included participation to international conferences, presenting the project through oral presentations and dissemination of advertising material. Furthermore, a plan of business development and a strategy for IP exploitation is under development, to boost the approaching of the project and its innovations toward the market.
A set of LED sources with corresponding excitation filters have been included within the optical path for sample fluorescence excitation under 3 well defined wavelengths. A highly sensitive fluorescence detector has been aligned on the output light path in order to collect fluorescence signals. This structure has been conceived to works in parallel with the proprietary laser-based technology developed by Foresee Biosystems. Though a fully automated 3-steps procedure (laser focus, laser auto-alignment and laser scan over the MEA electrodes), cardiac cells are simultaneously analysed both functionally and structurally. Two different objective lenses are exploited to focus laser and RGB light over the sample to provide high quality AP recordings (useful for analysis of defects in ion channels functionality) and sensitive fluorescence detection (useful for structural changes of cells).
In parallel, we worked on a novel enclosure case, including all the necessary electronic and mechanical support parts, focusing on the exploitation of material and fabrication approaches suitable for biomedical labs equipment.
A software graphical interface is under development. It allows for the parallel management of both laser scan procedure and fluorescence data acquisition from the detectors: a scientific CMOS camera equipped with sensitive sensors for fluorescence imaging and an industrial camera suitable for the Infrared imaging of the sample.
The software interface will include sub-libraries for filters and objective movements during the normal use, allowing an easy switch from the functional to the structural analysis of the samples.
In addition, a deep communication activity has been carried out to speed up SiMulTox outreach to the stakeholders. The main activity included participation to international conferences, presenting the project through oral presentations and dissemination of advertising material. Furthermore, a plan of business development and a strategy for IP exploitation is under development, to boost the approaching of the project and its innovations toward the market.
New tools and strategies in investigative toxicology are continuously improved in biotech companies to reduce safety-related attrition in drug development. Current toxicology assessment adopts a multi-level sequential approach, employing progressively in silico assays, biochemical and cellular in vitro assays and in vivo safety experiments, according to the progress of a drug candidate in the screening pipeline, to build a strong strategy of hazard identification and mitigation of potential risk on the humans. However, this pipeline still relies on the massive use of animal models which have different physiology from humans and, therefore, might hide alterations triggered by a drug treatment.
In the field of electro-active tissue model, such as cardiac or brain tissue, electrophysiological monitoring is a well-known and well accepted assay to investigate how a drug can affect the functionality these tissues.
Compared to current gold standard electrophysiology investigations, SiMulTox offers the unique advantage of performing simultaneously parallel functional and structural assays on the same sample. Due to its core technology, SiMulTox enables a new concept of multiparametric analysis of the very same sample, providing translationally relevant insights into drug effects. According to this new concept, long-term (chronic) toxicity on both function and morphology can be reliably assessed, providing more statistically relevant data with a lower number of samples. This leads to an unprecedent optimization of lab equipment, resources and efforts thus accelerating drug safety assessment and commercialization. All together, these capabilities follow the 3R rule (Replacement, Reduction, Refinement), which has been adopted recently to optimize the use of cellular methods instead of animal ones.
Starting from the next months, we will try to showcase the prototype of the platform for brief demonstration and first-hand experience to create awareness among the academic/industry researchers who might be interested in such a platform. In addition, further mechanical development will be implemented to finalize the first prototype, ready to a product validation in relevant environment (TLR5-6).
In the field of electro-active tissue model, such as cardiac or brain tissue, electrophysiological monitoring is a well-known and well accepted assay to investigate how a drug can affect the functionality these tissues.
Compared to current gold standard electrophysiology investigations, SiMulTox offers the unique advantage of performing simultaneously parallel functional and structural assays on the same sample. Due to its core technology, SiMulTox enables a new concept of multiparametric analysis of the very same sample, providing translationally relevant insights into drug effects. According to this new concept, long-term (chronic) toxicity on both function and morphology can be reliably assessed, providing more statistically relevant data with a lower number of samples. This leads to an unprecedent optimization of lab equipment, resources and efforts thus accelerating drug safety assessment and commercialization. All together, these capabilities follow the 3R rule (Replacement, Reduction, Refinement), which has been adopted recently to optimize the use of cellular methods instead of animal ones.
Starting from the next months, we will try to showcase the prototype of the platform for brief demonstration and first-hand experience to create awareness among the academic/industry researchers who might be interested in such a platform. In addition, further mechanical development will be implemented to finalize the first prototype, ready to a product validation in relevant environment (TLR5-6).