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.

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.

Naturally occurring post-transcriptional chemical modifications serve critical roles in impacting RNA structure and function. The combination of effects caused by modifications are ultimately linked to gene expression regulation at a genome-wide scale. Although examples of the importance of RNA modifications in translation are accumulating rapidly, still what these contribute to the function of complex physiological systems such as muscle is only recently emerging and addressed by our work.

This talk will explore effective strategies for the development and validation of qRT-PCR assays for PK studies of mRNA drug products. The presentation will cover key considerations in assay design, optimization, and validation processes to ensure accurate and reliable measurement of mRNA levels in clinical samples. Attendees can expect insights into methodologies aimed at enhancing the robustness and reproducibility of qRT-PCR assays in the evaluation of mRNA-based therapeutic products within clinical settings.

mRNA-based therapeutics offer treatments ranging from infectious disease to oncology to rare disease. Due to the rapid emergence of this technology, bioanalysts have needed to adapt bioanalytical approaches for monitoring pharmacokinetics, pharmacodynamics, and immunogenicity from the protein-based and gene-based therapies. In this presentation, we will discuss end-to-end bioanalytical support for mRNA-based therapeutics based on indication, as well as important life-cycle management considerations such as critical reagents.

Lipid nanoparticles have been used for the efficient delivery of different therapeutics. However, the LNP composition may be recognized by the immune system as foreign materials and activate the unintended immune response. In this talk, we will explore the immunogenicity risk of LNP composition, summarize the regulatory guidance and requirements for immunogenicity risk identification and evaluation, and provide immunogenicity assessment strategy for the novel mRNA/LNP therapeutics during drug development stage.

We demonstrate a fully synthetic method to prepare DNA and mRNA that is directly used as template for CFPS synthesis reactions using NEBExpress and PURExpress. Sequence fragments were assembled, amplified, and fed into a CFPS reaction. From the same DNA, mRNA template can also be prepared. We demonstrate the synthesis of several proteins, including toxic enzymes. These candidates could neither be expressed nor cloned in vivo. Our simple workflow allows the screening of multiple candidates, as well as large scale production of difficult proteins.

SRI International has developed an unbiased screening platform to identify peptides that mediate delivery throughout the CNS without disrupting the blood-brain barrier or destroying the biological cargo. This approach, named DiaCyt (Dia: Through and Cyt: Cell), utilizes phage-displayed libraries to identify ligands called molecular transport systems (MTS). Upon intravenous injection into a rat, the MTSs are transported into the CNS and is distributed throughout the ventricular system and within the surrounding parenchyma of the brain.

• Challenge issues in the bioanalysis of mRNA therapeutics with different technologies: CE, qRT-PCR, LC-MS et al.
• Impurity characterization for risk Assessment: impurity levels, criteria, and justification.
• Fit-for-purpose bioanalytical approaches to meet the needs of pharmacokinetics, pharmacodynamics, and immunogenicity assessment with mRNA therapeutics
• Product-related analysis to evaluate the immunological effects with mRNA therapeutics: protein expression from mRNA, infectious disease mRNA vaccine, and cancer mRNA vaccine.​

The traditional process of LNP development remains labor-intensive and cost-inefficient, relying heavily on trial and error. In this study, we present the AI-Guided Ionizable Lipid Engineering (AGILE) platform that streamlines the iterative development of ionizable lipids, crucial components for LNP-mediated mRNA delivery. This approach brings forth three significant features: efficient design and synthesis of combinatorial lipid libraries, comprehensive in silico lipid screening employing deep neural networks, and adaptability to diverse cell lines. This could significantly contribute to addressing the complex needs of mRNA delivery in clinical practice, thereby broadening the scope and efficacy of mRNA therapies.

This presentation will cover mRNA delivery approaches and technologies involved in transporting the nucleic acid to its target site to achieve the desired therapeutic effect, current challenges and opportunities in mRNA/LNP formulation development, and future directions for targeted mRNA delivery via LNP design and optimization.

This session on model-informed drug-development (MIDD) for mRNA therapeutics and vaccines explores the emerging field of utilizing mathematical modeling and data-driven approaches to optimize the development and dosing of innovative mRNA-based therapies and vaccines. Session includes topics including preclinical and clinical trial design, population pharmacokinetics/pharmacodynamics analyses, and systems pharmacology approaches for rational drug development. Discover the future of mRNA-based medicines in this evolving domain.

MIDD plays an important role in quantitative pharmacology by shifting the mindset to a “learning and confirming” paradigm to support informed decision-making. Large amounts of data are generated on a continuum in discovery and development that requires a holistic and systematic approach to curate results for evidence generation. This talk explores the different techniques and in silico tools that can be applied when developing a MIDD strategy for mRNA therapeutics across drug development. Advanced analytics including simple PKPD models to complex mechanistic QSP platform for LNP-encapsulated mRNA are reviewed with comparison to traditional clinical planning and design.