mRNAs As Medicines
Module 1: Introduction to Proteins and mRNAs
Course Introduction
This course, mRNAs as Medicines, is developed by Moderna and offered through Coursera. It's designed to provide a comprehensive understanding of mRNA medicines for everyone, regardless of their scientific background.
Module 1 Introduction
Module 1 sets the stage for understanding mRNA medicines by focusing on proteins and their crucial roles in the human body. We'll explore:
- The molecular composition of the human body
- Proteins and their functions
- Protein synthesis
- The role of proteins in viruses
Proteins and Their Roles in Human Health
Proteins are fundamental building blocks of all life forms, including humans. Here's what you'll learn:
- Proteins are everywhere: They constitute a significant portion of our body weight and are present in every cell.
- Diverse structures and functions: Proteins come in various shapes and sizes, performing structural, enzymatic, and transport roles. They are vital for maintaining our bodies' functions and overall health.
- Protein synthesis errors and diseases: Errors in protein synthesis can lead to various diseases. We'll examine Von Gierke disease as an example, where the inability to produce a functional version of a specific protein leads to severe health issues.
- Viruses and proteins: Viruses, though not technically alive, utilize proteins for their structure and to hijack host cells' protein synthesis machinery for replication.
Protein Structure
Delving deeper into proteins, this section covers:
- Amino acids: The building blocks of proteins, with 20 different types.
- Polypeptides and peptide bonds: Chains of amino acids form polypeptides through peptide bonds.
- 3D Structure determines function: The specific sequence of amino acids dictates how a protein folds into a unique 3D structure, which determines its function.
- Examples:
- Insulin: A small protein hormone crucial for regulating blood sugar levels.
- Human serum albumin: The most abundant protein in blood plasma, maintaining osmotic pressure, buffering pH, and transporting molecules.
- Hemoglobin: A multi-protein complex in red blood cells, responsible for oxygen transport.
mRNA Structure
With a solid understanding of proteins, we shift focus to mRNA:
- mRNA as blueprints: Messenger RNAs (mRNAs) act as blueprints carrying the genetic instructions for protein synthesis.
- Nucleotides and structure: mRNAs are composed of four nucleotides (A, C, G, U), and their sequence determines the protein's amino acid sequence.
- Visualizing mRNA: Atomic force microscopy (AFM) allows us to visualize the complex, flexible structures formed by mRNA molecules.
How Cells Make Proteins and Know What Proteins to Make
This section focuses on the intricate process of protein synthesis:
- Translation: The process of converting the genetic code in mRNA into proteins.
- Ribosomes: Tiny cellular machines that act as protein factories, reading mRNA and assembling amino acids.
- The Genetic Code:
- Codons: Three-letter sequences in mRNA that specify a particular amino acid.
- Start and Stop Codons: AUG signals the beginning of protein synthesis, while UAA, UGA, and UAG signal termination.
- Translation Process:
- Ribosome binds to mRNA.
- It moves along the mRNA, reading codons.
- tRNAs deliver corresponding amino acids.
- Amino acids are linked together to form a polypeptide chain.
- The process continues until a stop codon is reached.
- Multiple Ribosomes: Multiple ribosomes can simultaneously translate a single mRNA molecule, increasing protein production efficiency.
Where Do mRNAs Come From?
Unraveling the origin of mRNAs:
- Transcription: The process of creating an RNA copy from DNA.
- DNA as the Master Blueprint: Genomic DNA, housed within the cell nucleus, stores the genetic information for an organism.
- RNA Polymerase: The enzyme responsible for transcribing DNA into RNA.
- Central Dogma of Biology: The flow of genetic information from DNA to RNA to protein.
Module 1 Summary
Module 1 provides a foundational understanding of proteins and mRNAs, their structures, synthesis processes, and importance in biological systems, laying the groundwork for understanding mRNA medicines.
Module 2: Learning About Medicines
Module 2 Introduction
Building upon the knowledge of proteins and mRNAs, Module 2 delves into the world of medicines. We'll explore:
- What defines a medicine.
- Different types of medicines (small molecules and protein biologics).
- How proteins and mRNAs can be utilized as medicines.
Medicines: Small Molecules and Proteins
This section examines traditional medicines and paves the way for understanding mRNA-based approaches:
- Defining "Medicine": A substance used for diagnosis, cure, treatment, or prevention of diseases.
- Small Molecule Medicines:
- Characteristics: Small size, often taken orally, work by modulating protein activity.
- Examples: Antibiotics (penicillin), painkillers (ibuprofen, aspirin).
- Limitations: Drug development is time-consuming, expensive, and has a high failure rate.
- Protein Medicines (Biologics):
- Therapeutic Proteins: Proteins used as medicines, mostly functioning in the bloodstream.
- Examples: Insulin (diabetes), clotting factors (hemophilia), erythropoietin (anemia), monoclonal antibodies ( various diseases).
- Limitations: Production challenges, potential for immune reactions, limited cell penetration.
What Makes mRNA Suitable as a Medicine?
This section explores the characteristics of mRNA that make it a promising candidate for medicine:
- Ideal Medicine Properties:
- Dose-dependent effect
- Time-limited effect
- Repeatable dosing
- mRNA as a Transient Molecule: mRNA and the proteins they encode have limited lifespans, allowing for controlled and adjustable therapeutic effects.
- Benefits of mRNA Medicines:
- Targeting any protein: Unlike traditional protein therapies, mRNA can target proteins inside cells, expanding therapeutic possibilities.
- Simplified manufacturing: mRNA medicines are manufactured through a cell-free process, potentially allowing for faster and more cost-effective production.
- Platform technology: The mRNA platform can be adapted to target different diseases by simply changing the mRNA sequence, simplifying development.
- Multiplexing: Multiple mRNAs can be delivered simultaneously, enabling the production of multiple therapeutic proteins.
Making and Delivering mRNA Medicines
Moving from theory to practice, this section delves into the practical aspects of mRNA medicine development:
- mRNA Synthesis:
- DNA Template: A DNA template containing the desired mRNA sequence is used.
- In vitro Transcription: The DNA template is transcribed into mRNA using enzymes.
- Purification and Quality Control: The synthesized mRNA undergoes rigorous purification and quality checks.
- Delivery Challenges:
- Degradation: Naked mRNA is rapidly degraded in the body.
- Targeting: mRNA lacks inherent mechanisms for tissue-specific targeting.
- Cell Entry: The negatively charged mRNA faces challenges crossing cell membranes.
- Lipid Nanoparticles (LNPs) as Delivery Vehicles:
- Protection: LNPs encapsulate and protect mRNA from degradation.
- Targeting: LNPs can be engineered for tissue and cell-specific delivery.
- Cell Entry: LNPs facilitate mRNA entry into cells.
Anatomy of a Lipid Nanoparticle (LNP)
This section provides a detailed look at LNPs, the workhorses of mRNA delivery:
- Structure and Composition:
- mRNA Cargo: The therapeutic mRNA molecule.
- Ionizable Lipids: Interact with and encapsulate the mRNA.
- Phospholipid Bilayer: Forms a stable outer shell.
- Cholesterol: Provides structural integrity.
- Surfactant (PEG): Prevents LNP aggregation.
- Size and Appearance: Tiny spherical structures, 80-100 nanometers in diameter, visible under an electron microscope.
- Mechanism of Action:
- LNP-mRNA complexes are taken up by cells through endocytosis.
- LNPs escape the endosome and release mRNA into the cytoplasm.
- mRNA is translated into the therapeutic protein.
- Fate of LNP Components: LNP components are naturally metabolized and cleared by the body.
mRNA Medicines and the Innate Immune System
A crucial aspect of mRNA medicine development is navigating the body's immune system:
- Innate vs. Adaptive Immunity:
- Innate Immune System: First line of defense, recognizes general pathogen features, rapid response.
- Adaptive Immune System: Highly specific, develops over time, involves B cells (antibodies) and T cells (cellular immunity).
- Innate Immune Sensors and mRNA: The innate immune system has sensors that detect foreign RNA, potentially triggering an immune response against mRNA medicines.
- Overcoming Immune Activation:
- Modified Nucleotides (e.g., 1-methylpseudouridine): Replacing uridine with modified nucleotides reduces immune stimulation.
- Minimizing Double-Stranded RNA: Optimizing mRNA production to reduce double-stranded RNA byproducts, which can trigger an immune response.
Module 2 Summary
Module 2 explores the world of medicines, highlighting the limitations of traditional approaches and the potential of mRNA as a new therapeutic modality. It details the properties that make mRNA suitable for medicine, the challenges in making and delivering mRNA medicines, and the strategies employed to overcome these challenges.
Module 3: Applications for mRNA Medicines
Module 3 Introduction
In Module 3, we'll explore the diverse applications of mRNA medicines in both preventing and treating diseases:
- mRNA vaccines: Preventing infectious diseases.
- mRNA therapeutics: Treating existing diseases, including cancer and genetic disorders.
- Regenerative medicine: Harnessing mRNA for tissue repair and regeneration.
Prophylactic mRNA Vaccines - Part 1
This section dives into the groundbreaking world of mRNA vaccines:
- Prophylactic Vaccines: Vaccines designed to prevent diseases.
- Traditional Vaccines:
- Inactivated Pathogen Vaccines: Use killed pathogens (e.g., polio, typhus vaccines).
- Attenuated Pathogen Vaccines: Use weakened pathogens (e.g., MMR vaccine).
- Limitations: Potential for incomplete inactivation, risks for immunocompromised individuals.
- How Vaccines Work:
- Antigen Presentation: Antigen-presenting cells (APCs) engulf pathogens, process them, and display fragments (antigens) on their surface.
- T Cell and B Cell Activation: APCs activate T cells and B cells that recognize the specific antigen.
- Antibody Production and Cellular Immunity: B cells produce antibodies, while cytotoxic T cells target infected cells.
- mRNA Vaccines:
- mRNA Delivery: LNPs deliver mRNA encoding viral proteins into APCs.
- Protein Production: APCs produce viral proteins using the delivered mRNA.
- Antigen Presentation: APCs display the produced viral proteins on their surface, triggering an immune response.
Prophylactic mRNA Vaccines - Part 2
Continuing the discussion on mRNA vaccines, this section delves into their advantages and limitations:
- Advantages of mRNA Vaccines:
- Multiplexing:
- Delivering multiple mRNAs in a single vaccine.
- Enabling the production of multi-protein complexes (e.g., CMV vaccine).
- Creating combination vaccines against multiple pathogens (e.g., bivalent COVID- 19 vaccines, potential for RSV, COVID-19, and flu combination vaccines).
- Resource Efficiency:
- Lower resource requirements compared to traditional vaccines (e.g., no need for eggs for flu vaccines).
- Smaller manufacturing footprint.
- Rapid Production: Significantly faster production times compared to traditional vaccines.
- Decentralized Manufacturing:
- mRNA sequence information can be easily shared, enabling distributed manufacturing.
- Potential for rapid response to outbreaks and pandemics.
- Multiplexing:
- Limitations of mRNA Vaccines:
- mRNA Fragility:
- Early mRNA vaccines required ultra-cold storage.
- Ongoing research to improve stability (e.g., lyophilization, microneedle patches).
- Maximum Tolerated Dose:
- The number of different mRNAs in a vaccine is limited by the maximum tolerated dose of LNPs.
- Requires careful optimization of mRNA ratios in multiplexed vaccines.
- mRNA Fragility:
mRNA Therapeutics that Stimulate the Immune System
This section focuses on mRNA therapeutics that activate the immune system to fight diseases:
- Immunogenic mRNA Therapeutics: Similar to vaccines, they stimulate an adaptive immune response.
- Target: Not external pathogens, but internal threats like cancer cells.
- Individualized Neoantigen Therapies (INTs) for Cancer:
- Neoantigens: Unique proteins produced by cancer cells due to mutations.
- INT Mechanism:
- Neoantigen Identification: Sequencing tumor DNA to identify neoantigens.
- mRNA Design: Designing mRNAs encoding the identified neoantigens.
- mRNA Delivery: Delivering mRNAs to APCs.
- Immune Activation: APCs present neoantigens to the immune system, triggering an attack on cancer cells.
- INTs as Personalized Cancer Vaccines: INTs are tailored to an individual's specific tumor mutations, potentially offering more targeted and effective cancer treatment.
Non-Immunogenic mRNA Therapeutics
In contrast to immunogenic therapies, this section covers mRNA therapeutics that do not rely on immune stimulation:
- Non-Immunogenic mRNA Therapeutics: Deliver mRNAs encoding therapeutic proteins to replace missing or defective proteins, particularly for genetic diseases.
- Protein Replacement Therapies:
- Addressing Inborn Metabolic Diseases:
- Many genetic disorders are caused by the inability to produce specific enzymes.
- mRNA therapies can deliver the missing enzyme's blueprint directly to cells.
- Example: Von Gierke disease (glucose-6-phosphatase deficiency).
- Challenges and Considerations:
- Targeting: Efficiently delivering mRNA to the relevant organ (e.g., the liver for many metabolic disorders).
- Dosing: Larger doses may be required compared to vaccines.
- Chronic Dosing: Repeated administration is necessary for long-term treatment.
- Addressing Inborn Metabolic Diseases:
- Other Applications of Non-Immunogenic mRNA Therapeutics:
- Cystic Fibrosis: Delivering mRNA encoding the CFTR protein to lung cells.
- Regenerative Medicine:
- Stimulating blood vessel growth (e.g., VEGF for heart muscle).
- Repairing damaged tissues (e.g., growth factors for urinary incontinence).
- Secreted Protein Production:
- Inducing the body to produce therapeutic monoclonal antibodies.
- Potential for treating viral infections and other diseases.
Module 3 Summary
Module 3 showcases the diverse applications of mRNA medicines, from preventing infectious diseases with prophylactic vaccines to treating cancer with individualized neoantigen therapies and addressing genetic disorders with protein replacement therapies. It highlights the ongoing research and development in this rapidly evolving field, emphasizing the potential of mRNA to address various unmet medical needs.
Course Conclusion
This course provides a solid foundation in understanding mRNA medicines, their mechanisms, applications, and potential to transform healthcare. As research and development progress, we can anticipate exciting advancements and broader applications of this innovative technology in the future.
