Applications of mRNA Therapeutics

To reach the full potential of the disease-agnostic RNA platform, we are developing a set of synergistic technologies consisting of continuous manufacturing processes (continuous enzymatic RNA synthesis, continuous downstream purification, and continuous lipid nanoparticle formulation), analytical technologies, computer models, and bespoke software. These technologies are co-developed under a patient-centric Quality by Digital Design (QbDD) framework to obtain a multi-product design space with CQAs as a function of CPPs, and aid process automation. By combining the QbDD framework with the RNA platform, vaccines and therapeutics can be developed and mass-produced faster against a wide range of diseases at low cost.

Our work is focusing on ensuring the equitable access to RNA based vaccines by everyone, everywhere. This vision is underpinned by the creation of flexible manufacturing networks comprising autonomous and multipurpose RNA manufacturing and discovery nodes in any geography. In this talk, a brief overview of the BiaB technology will be given, focusing then on our purification methods and our Quality-By-Design framework with examples.

mRNA-based vaccines use lipid nanoparticles (LNPs) to ensure mRNA delivery for effective protein translation. However, the large size and unique behavior of the LNPs create significant challenges in the design and implementation of the sterile filtration step. Filter capacities can be significantly smaller than those for traditional biotherapeutics, and the fouling behavior does not follow traditional mechanisms. The results provide important insights into the mechanisms controlling sterile filtration of LNPs.

A rapid, flexible, and affordable mRNA manufacturing workflow is necessary to unlock the full potential of mRNA therapeutics. mRNA transcription relies on the rigid sequence design of constructs obtained by laborious bacterial cloning and time-consuming plasmid preparation. Here, we introduce a synthetic DNA template (SDT)-based platform, allowing cost, and time-efficient production of mRNA molecules encoding different protein candidates for therapeutic evaluation, enabling applications such as T cell receptor (TCR) screening, and neoantigen immunogenicity validation directly from tumor patients. The presentation will cover therapeutic evaluation, immunogenicity investigation, and mRNA molecular characterisation.

Immunotherapy, particularly mRNA therapeutics, is redefining cancer treatment by harnessing the body's immune system to target cancer cells. These therapies, delivering tumor-specific antigens or immune modulators via mRNA, are enhancing immune recognition and attack on cancer cells. However, challenges like limited patient response due to inadequate T cell activation in the tumor microenvironment persist. Addressing this, we present STX-001, a novel synthetic self-replicating mRNA technology for intratumoral delivery. It provides sustained IL-12 cytokine expression, specifically within the tumor microenvironment, improving T cell recruitment and activity. STX-001 exemplifies the next step in personalized immunotherapy, combining the precision and adaptability of mRNA therapeutics to overcome existing immunotherapy limitations.

The National Center for Therapeutics Manufacturing (NCTM) partnered with the Biomedical Advanced Research and Development Authority (BARDA), Pfizer, the US Pharmacopeia (USP), and other solution providers to develop the first commercial-scale, hands-on training course for mRNA vaccine production in the US. This presentation will give an overview of the company-agnostic, in-house platform process that was developed for the production of mRNA-LNPs, beginning with pDNA production and continuing through bulk filling/freezing.

This presentation focuses on the strategy we take at Korro Bio on building a platform that harnesses the body’s natural RNA editing machinery, ADAR (Adenosine deaminases acting on RNA) mediated RNA editing, to treat previously untreatable diseases and the potential of this platform in broad applications.

A/H2N3 influenza has posed a particular pandemic threat. Self-amplifying mRNA (sa-mRNA) vaccine emerges as the counter measurement. We evaluated sa-mRNA A/H2N3 vaccine designs, including codon optimization and co-expression of neuraminidase (NA), in addition to hemagglutinin (HA). Mouse studies showed robust neutralizing antibody and CMI responses to both HA and NA, and strong cross-reactivity to another A/H2N3 virus, demonstrating the sa-mRNA pandemic vaccine as the effective approach for pandemic preparation.

A multiclade env+gag+pro mRNA vaccine capable of producing virus-like particles (VLP) in vivo induced broad-spectrum neutralization, albeit at low titers, and partial protection in nonhuman primates. Is an HIV-1 vaccine feasible? Why is it so challenging to develop an HIV-1 vaccine? Why is mRNA especially suited as a vehicle for an HIV-1 vaccine? What are the requirements for the induction of broadly neutralizing antibodies? Is protection from HIV-1 possible without the induction of broadly neutralizing antibodies?

Since the discovery of circular RNA, a new class of single-stranded RNA, their biogenesis, regulation, and function have been rapidly characterized, allowing for better understanding and their adoption as new tools for therapeutic applications. With the development of biotechnology and molecular medicine, circRNAs have been engineered as a novel class of RNA therapeutics. In the field of vaccines, compared to linear mRNA vaccines, mRNA vaccines offer an improved approach to RNA-based vaccination with increased stability, simplicity of manufacture, and scalability.

In order to exploit the potential of mRNA technology for pharmaceutical application, tailored delivery systems are required. NeoVac has a pioneering 2.0 LNP Platform for superior delivery of RNA payloads for vaccines and therapeutic intervention, such as treatment for metabolic diseases, oncology treatments, genetic diseases, and others, based on novel, proprietary lipids. The technology allows to one specifically target different organs and to modulate immunogenicity according to the intended intervention. Here we present first data to underline the potential of our new technology.

Modified mRNA (modRNA) technology, lauded for its triumphs in COVID-19 vaccine development, is emerging as a promising strategy against cardiovascular diseases (CVD). With 19.1 million global deaths in 2020 and a prevalence of 620 million, CVD demands innovative solutions. Despite medical strides, the lack of a cure intensifies public health concerns. My presentation spotlights our work on modRNA therapies fostering cardiac regeneration and combating cardiac fibrosis, hypertrophy, and cell death in CVD animal models. We also showcase our cell-specific modRNA expression platforms, contributing to evolving modRNA therapeutic landscapes for CVD.

In vivo cellular reprogramming via targeted delivery of RNA-based therapeutics to selective cells could be highly valuable. In this talk, I will discuss some of our recent efforts in developing targeted mRNA therapeutics for non-oncology indications.