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.
Team
Rajesh V Iyer
Lysosomal storage disorders (LSDs) are a group of monogenic rare genetic disorders that occur owing to a loss of lysosomal enzyme function. In LSD patients, intravenous administration of the functional enzyme i.e., enzyme replacement therapy (ERT) has been found to rescue disease symptoms and improve the patient’s life. Though effective to a certain degree, ERT-based therapies are very costly. Globally, LSD prevalence is taken to be 1 in 7000-8000 individuals and if extrapolated, there could be more than 1 lakh LSD patients in India. More than 95% of the Indian population cannot afford these drugs. This cost is primarily due to the small market size, cost-intensive manufacturing, and purification methods to produce therapeutic proteins. The LSD patients require ERT for their entire life and owing to the absence of any indigenous drugs, very few Indian LSD patients can access these drugs via humanitarian funds. Many succumb to the disease, often due to the unavailability of these drugs.
To address this emergency, 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 solutions for many diseases
Investigator: Rajesh V Iyer
GNE myopathy is an adult-onset progressive neuromuscular disorder that generally leads to extreme disability in a few years. The disease is caused by mutations in the bifunctional enzyme Glucosamine (UDP-N-Acetyl)-2-Epimerase/N-Acetyl-Mannosamine Kinase involved in sialic acid biosynthesis. There is no approved treatment for this disease.
TIGS is collaborating with World Without GNE Myopathy (WWGM), a patient advocacy group, to develop mRNA therapy for GNE Myopath. Currently, the mRNA team at TIGS has generated two GNE-encoding mRNA candidates and tested them using cell culture methods. The GNE-mRNA candidates can express in cell lines for more than 4 days and efforts are now on developing muscle targeting nanoparticle formulations.
Investigator: Rajesh V Iyer
We have generated mRNA-lipid nanoparticles (mRNA-LNPs) using a customizable microfluidic device. The size of mRNA-LNPs was found to be below 100 nanometres with a net negative surface charge. These mRNA-LNPs were highly efficient in transfecting mRNA encoding the green fluorescent protein into HEK 293 cells. Upon microscopic examination, we found that under in vitro conditions, more than 95% efficiency of HEK 293 transfection can be achieved by using 250 nanograms of mRNA-LNPs.
Investigator: Rajesh V Iyer