mRNA Therapeutics

Messenger RNAs (mRNAs) are a fast-emerging class of biotherapeutics. mRNA therapies offer a new opportunity for targeted treatment of challenging diseases and flexible manufacturing, as demonstrated by the rapid development of mRNA vaccines against COVID-19. They are non-infectious, non-integrating, and cell-free, offering both rapid and readily scalable production with high productivity. Our team at TIGS has begun working towards improving the purification of synthesized mRNA and developing alternative lipid formulations for improved encapsulation and stability, using specialized devices for encapsulation and high throughput assessment of lipid formulations.

Monogenic rare genetic disorders are a result of loss of protein function. In a few such diseases, intravenous biweekly administration of therapeutic protein has been found to rescue disease symptoms and improve the patient’s life. Though effective to a certain degree, these therapies are costly. This cost is primarily due to the cost intensive purification methods to produce therapeutic proteins.

We reasoned that, as an alternative to protein therapy, mRNA encoding therapeutic proteins can be utilized to produce the therapeutic proteins in vivo. mRNA production, owing to its synthetic nature, is highly scalable with a relatively smaller footprint which ultimately leads to affordable therapeutic solution for many diseases. We will begin with the mRNA-LNP particle production and test it on relevant disease models available at TIGS. We have already standardized the mRNA synthesis, purification of mRNA using different kits, capping and polyA tailing.

This will be followed by setting up a complete mRNA therapy lab including nanoparticle synthesis equipment and particle analyzer as well as the cell culture, mRNA transfections and expression analysis systems needed.

Rajesh V Iyer

We look to generate encapsulated complexes of mRNA encoding a therapeutic protein and to validate mRNA complexes in disease-specific cell lines/patient derived cell lines, wild type murine inbred lines and in disease mouse models.

Based on this we have started work on mRNA- based therapeutic candidates for lysosomal storage disorders viz. Pompe Disease (GAA deficiency), Fabry Disease (GLA deficiency) and Gaucher Disease (GBA deficiency)

Investigator: Rajesh V Iyer

Ashoka University, Sonipat
National Institute of Mental Health and Neurosciences
ICAR – National Institute of Veterinary Epidemiology and Disease Informatics (NIVEDI)
CSIR-Institute of Genomics and Integrative Biology (IGIB)
Indian Institutes of Science Education and Research, Tirupati
DBT-Institute for Stem Cell Science and Regenerative Medicine (DBT-InStem)

The aim of the project is to develop stably expressing SMN-mRNA based neuron-targeting lipid nanoparticles. The following is being done to achieve it.

• Set-up cell culture for evaluating expression from different mRNA constructs.
• Test different UTRs to identify most stable ones.
• Generate neuron-targeting lipid nanoparticles.

The idea of the project is to develop Cas9 and Cas12 mRNA-LNP for genome editing in mammalian cell lines.

• Design Cas9 and Cas12 mRNA with UTRs and NLS for optimum expression
• Test the expression of the IVT mRNA using HEK293 cell lines.
• Validate genome editing activity using pre-tested gRNA.
• Test the Cas12 mRNA with candidate gRNAs for reversing Rare disease mutations.