Fraunhofer Institute for Toxicology and Experimental Medicine
Fraunhofer Institute for Toxicology and Experimental Medicine

RNA technologies

Developing RNA-based compounds to the stage of clinical use

RNA for targeted therapies

Um die molekularen Signalwege in gesunden und kranken Herzen besser zu verstehen, analysieren Forschende nicht-codierende Mikro-RNAs mithilfe der Real-time-PCR. Mikro-RNAs, die im Zusammenhang mit der Krankheit stehen, wie etwa die Mikro-RNA-132, können somit identifiziert werden.
© Fraunhofer ITEM, Ralf Mohr
In order to better understand the molecular signaling pathways in healthy and diseased hearts, researchers analyze non-coding microRNA molecules by real-time PCR.

RNA-based therapeutics are a drug class with huge potential for medicine — and scientists have barely scratched the surface of their possible use. Fraunhofer ITEM researchers develop novel drugs and methods for RNA-based therapeutic concepts.

The targeted use of RNA compounds based on coding or non-coding RNA sequences enables a tailored response of the respective target cells under certain pathological conditions. Fraunhofer researchers use a wide range of RNA-based compounds, such as small interfering RNA, nucleoside-modified messenger RNA or RNA blockers — from target discovery through to preclinical development programs. Bioinformatic models play a key role when it comes to selecting disease-associated RNAs, studying their interaction with other genes, and optimizing drug design.

Special viral and non-viral administration technologies, both at the molecular and equipment level, are being developed for the targeted use of RNA compounds. Local delivery via the airways is a promising solution in particular for RNA therapeutics used to combat lung diseases.

Safety and efficacy testing of RNA compounds

For safety and efficacy testing of therapeutics, researchers use proven in-vitro and in-vivo animal models and are continuously developing new model systems that are based on human tissues as well as mini organs and cells derived from stem cells. 

RNA molecules as biomarkers

RNA molecules are attracting increasing interest for diagnostic purposes as well. The expression profiles of RNA molecules are altered in many pathologies, such as lung diseases and cancer. RNA can be obtained from various liquid biopsies, such as blood, urine or cerebrospinal fluid. Taking a liquid biopsy is much less invasive for patients than a tissue biopsy. Next generation sequencing technologies allow the RNA composition of a sample to be determined. RNA expression profiles can thus be used as biomarkers to characterize diseases, predict the response to a specific therapy, and monitor treatment success. Fraunhofer ITEM researchers have developed technologies that enable the analysis of even very small quantities of RNA as well as determination of the expression profiles of single cells or cell-free, circulating RNA.

RNA molecules as biomarkers
© Fraunhofer ITEM, Ralf Mohr
So far, mainly mRNA and miRNA expression profiles have been used as biomarkers, although circular RNA (circRNA) is also emerging more and more as a biomarker with great potential.

RNA research: recent projects and highlights

 

© duyina1990 KI, adobe.stock.com

Cardiac fibrosis: safety assessment of a novel therapeutic approach

ITEM researchers evaluated the tolerability of a drug candidate for heart failure in safety studies and determined a NOAEL.

READ MORE
 

© Bundesagentur für Sprunginnovationen

Project iGUARD wins the SPRIN-D Challenge "Broad-Spectrum Antivirals"

Researchers in the iGUARD project are developing RNAi inhalation therapies that can be quickly adapted to different respiratory viruses.

READ MORE
 

© Ekaterina_1525 - stock.adobe.com / Fraunhofer IZI / Alernon77 - stock.adobe.com / CoreDESIGN - stock.adobe.com / Fraunhofer IZI

Project RNAuto

Fraunhofer develops automated production technologies for mRNA-based drugs.

Read more

Circular RNA: an innovative RNA therapy agent?

Read More

Project archive

Here you can find more projects sorted by our research and development competences. 

Our technologies and methods

Expand all Close all

Identification of target structures

© Fraunhofer ITEM, Ralf Mohr

Computer-based models can provide clues to molecular RNA targets in different human organ pathologies.

Based on these structures, RNA drug candidates can be synthesized and subsequently validated in translational model systems.

Lead optimization

Researcher in the laboratory.
© Fraunhofer ITEM, Ralf Mohr

Analytical methods are essential for the optimization of RNA-modulating agents. Oligonucleotide-based binding assays are used to validate the structure of the active ingredient in advance and adjust it accordingly.

The ensuing efficacy testing and pharmacokinetics studies are also performed with the help of bioanalytical methods. Microscale thermophoresis (MST) and HPLC platforms are available as well.

 

Efficacy testing in human model systems and RNA-based analytical methods (PK/PD studies)

Hand holding a cultivation chamber with a cardiac tissue section using tweezers.
© Fraunhofer ITEM, Ralf Mohr

Human and animal tissue culture models

Heart

  • Cell cultures
  • Cardiac tissue slices
  • 3D organoid models

Lung

  • Cell cultures
  • Isolated perfused lung
  • Precision-cut lung slices (PCLS)
  • 3D organoid models

Liver

  • Cell cultures
  • Precision-cut liver slices(PCLiS)

Toxicology (in vitro/in vivo)

  • Preclinical testing in vivo
  • Preclinical tests using different in-vitro and ex-vivo lung and heart models
  • Safety pharmacology
  • Toxicological risk assessment
    • Identification and use of predictive biomarkers for toxicological endpoints
    • Generation of mechanistic data to support next generation risk assessment
    • Assessment of genotoxic impurities according to ICH M7, including review of literature, QSAR analysis, and PDE derivation

 

Female researcher working at a sterile workbench.
© Fraunhofer ITEM, Ralf Mohr

RNA targeted delivery

Different technologies and cardiopulmonary 2D and 3D model systems are used to develop the targeted, organ-specific delivery of RNA therapeutics.

  • Bioanalytics for therapeutic RNA interaction with the target sequence
  • Manufacturing and validation of nanoparticles as carriers of RNA therapeutics
  • Biological validation of RNA-nanoparticle complexes
  • Development of cell-specific nanoparticles
  • Nebulization of RNA-nanoparticle complexes
Index finger of a hand in a laboratory glove taps on a laboratory device.
© Fraunhofer ITEM, Ralf Mohr

Clinical trials phases I/II

The participant is examined and informed by the study physician.
© Fraunhofer ITEM, Ralf Mohr
Proband schüttelt Studienärztin die Hand vor einer Untersuchung.
  • Phase-I unit and infrastructure for clinical research
  • Broad range of diagnostic methods for safety and efficacy assessment
    • MRI to visualize lung ventilation and blood flow
    • ECG, echocardiography, and telemetry
  • Controlled aerosol inhalation
  • Broad range of challenge models for early proof of concept

Types of RNA

There are many different types of RNA. The most widely known is messenger RNA (mRNA). It carries the genetic information needed to produce proteins, such as for the SARS-CoV-2 spike protein in coronavirus vaccines. But in addition to mRNA, a large number of other RNA molecules are present in the organism. These are referred to as non-coding RNAs, including, for example, microRNA (miRNA), long non-coding RNA (lncRNA), and circular RNA (circRNA), and they play essential regulatory roles inside cells. Their dysregulation can result in a broad range of diseases, which is why they are crucial therapeutic targets.

© CROCOTHERY, adobe.stock.com

Messenger RNA

 

Messenger-RNA (mRNA) is a single-stranded ribonucleic acid that carries and transmits genetic information for building a certain protein inside a cell.

mRNA-based drugs introduce the blueprint for a particular protein into cells and the cells will then synthesize this protein. In the case of mRNA vaccines, the synthesized protein serves as antigen, triggering an immune response to build protection.

© CROCOTHERY, adobe.stock.com

MicroRNA

 

MicroRNA is a class of highly conserved, short, non-coding RNAs that play a crucial role in the complex network of gene regulation.

The various miRNA molecules regulate gene expression with a high degree of specificity at the post-transcriptional level and can thus serve as therapeutic targets.

© CROCOTHERY, adobe.stock.com

Small interfering RNA

 

The term small interfering RNA (siRNA) refers to synthetically produced RNA molecules that are used in research, but also as therapeutic agents. siRNA molecules with any desired sequence can be produced by chemical synthesis. 

To introduce siRNA into cells, a method known as liposomal transfection or lipofection is mainly used. It couples RNA with liposomes that fuse with the target cell membrane, thereby delivering the siRNA into the cell.

© CROCOTHERY, adobe.stock.com

Circular RNA

 

Circular RNA (circRNA)  is a type of single-stranded ribonucleic acid which, unlike linear RNA, forms a covalently closed continuous loop. Because circular RNA does not have 5' or 3' ends, it is resistant to exonuclease-mediated degradation and is presumably more stable than most linear RNA in cells.

circRNA is involved in gene regulation and has been linked to some diseases such as cancer. The biological function of most circRNA molecules is as yet unclear.

© CROCOTHERY, adobe.stock.com

IncRNA

 

Long non-coding RNAs (lncRNA) are generally defined as transcripts with more than 200 nucleotides that are not translated into protein.

They are involved in gene regulation at the transcriptional, post-transcriptional and epigenetic levels and have an impact on DNA replication timing and chromosome stability.

© CROCOTHERY, adobe.stock.com

modRNA

 

Nucleoside-modified messenger RNA (modRNA) is a synthetic, chemically modified mRNA, in which some nucleosides have been replaced by other naturally modified nucleosides or by synthetic nucleoside analogues.

Such RNA molecules are used in experimental or therapeutic contexts to induce the production of a desired protein in certain cells.