Cell and Gene Therapy

Plasmid DNA and Viral Vector Applications and Discovery Services

Cell and gene therapies are transformative for the future of medicine and patient success, particularly for treating diseases that currently have limited or no current treatment options. They hold the potential for personalized, durable, and even curative outcomes.

What exactly are cell and gene therapies? We are glad you asked! 

Cell and gene therapies are incredibly advanced medical treatments designed to treat or potentially cure diseases by addressing the underlying genetic causes or by enhancing cellular functions by introducing or inducing optimized cellular behavior. This medical advancement has many applications and different types of patient outcomes because of how limitless the effects of gene editing can have on patients.

Cell Therapy

  • Regenerative Medicine: Cell therapy is used to replace or repair damaged or diseased cells and tissues from that may occur from inherited diseases, serious injury, or infectious diseases. For example, stem cell therapies are applied in regenerative medicine to treat conditions like heart disease, spinal cord injuries, and certain types of blood disorders. Read more about plasmid DNA and viral vectors in regenerative medicine
  • Immunotherapy: Cells such as T-cells are modified or engineered to target and destroy cancer cells. CAR-T cell therapy is a key example, where a patient’s own T-cells are engineered to recognize and attack cancer cells. Read more about CAR-T and lentivirus applications further down.
  • Tissue Engineering: Cells can be used to create tissues or even organs that can be transplanted into patients. This approach could address organ shortages met through bioprinting or treat conditions like severe burns or degenerative diseases.

Gene Therapy

  • Correcting Genetic Disorders: Gene therapy involves introducing, removing, adding, silencing or altering in other ways, genetic material within a patient’s cells to treat diseases caused by genetic mutations. This is particularly applicable for monogenic disorders like cystic fibrosis, hemophilia, or sickle cell anemia.
  • Oncology: Gene therapy can be used to introduce new genes, through viral and non-viral vectors, into cancer cells to either kill them or make them more susceptible to other treatments like chemotherapy or radiation. Learn more about plasmid DNA and viral vectors in oncology.
  • Vaccination and Preventive Therapies: Genetic material can be introduced into the body to stimulate an immune response, similar to vaccines. 

The real question is how cell and gene therapies can help patients recover from disease, improve the quality of their life, and ultimately have better outcomes. The ultimate patient outcome depends on what specific illness, ailment, or preventative measure, the cell or gene therapy is targeting.

  • Curative Potential: Unlike traditional treatments, cell and gene therapy often aim for a one-time or limited-duration treatment that could result in a long-term cure. For example, patients with inherited genetic diseases may have their faulty genes corrected, potentially curing the condition.
  • Target Cells: Used to transduce host T cells. Once modified, these T cells can recognize and attack cancer cells.
  • Application: Involves extracting T cells from a patient, modifying them with the CAR-T lentivirus, and then reintroducing them into the patient to fight cancer.

Plasmid DNA in Cell and Gene Therapy Molecular Cloning and Mutagensis

These molecular biology techniques, molecular cloning and mutagenesis, are integral to creating and modifying the genetic material in plasmid DNA used in these therapies for the following reasons:

  • Gene Delivery: In gene therapy, plasmid DNA is used to deliver therapeutic genes into target cells. These genes can correct a defective gene, produce a therapeutic protein, or regulate gene expression in the patient’s cells.
  • Viral Vector Production: Plasmids are used to produce viral vectors (such as AAV or lentivirus) that are widely used to deliver therapeutic genes in gene therapy. Plasmid DNA is transfected into producer cells, which are then used to package viral vectors containing the therapeutic gene.

Molecular Cloning in Cell and Gene Therapy:

  • Therapeutic Gene Cloning: Molecular cloning allows scientists to insert a therapeutic gene (or other genetic elements) into a plasmid vector. This cloned plasmid can then be amplified in bacteria and purified for further use in therapy.
  • Optimization of Gene Expression: Molecular cloning techniques are used to fine tune plasmids to optimize the expression of therapeutic genes, making sure they are efficiently expressed in the target cells. Researchers can clone different promoters, enhancers, or regulatory sequences into plasmids to achieve the desired expression levels.
  • Customizing Plasmid Vectors: Researchers can clone multiple genetic elements into plasmids, including therapeutic genes, reporter genes (such as GFP), and selection markers, allowing for tailored solutions in gene therapy. These customized plasmids are often used in viral vector production for gene delivery.

Mutagenesis in Cell and Gene Therapy

  • Engineering Therapeutic Proteins: Mutagenesis is used to alter specific amino acids in therapeutic proteins, improving their stability, efficacy, or reducing immunogenicity. For instance, mutagenesis can optimize antibodies or enzymes used in gene therapy.
  • Studying Disease Models: By introducing specific mutations into genes (via plasmid DNA), researchers can create models of genetic diseases, allowing them to study disease mechanisms and test potential therapies.
  • Optimizing Gene Therapy Vectors: Mutagenesis can be applied to optimize the components of gene therapy vectors, such as improving viral capsid proteins for better targeting and delivery, or modifying regulatory elements to optomize gene expression.

Plasmid DNA Based Cell and Gene Therapy Companies and Therapeutic Candidates

  • Pfizer: Comirnaty: The COVID-19 mRNA vaccine used plasmid DNA during the manufacturing process to generate the mRNA template. Although this isn’t a direct gene therapy, plasmid DNA is crucial in the production process.
  • Poseida Therapeutics: P-BCMA-101: A CAR-T cell therapy targeting multiple myeloma. Poseida uses a proprietary piggyBac transposon system, where plasmid DNA is used to modify T-cells without viral vectors, making this a non-viral gene therapy approach.
  • Genprex: Reqorsa: A gene therapy candidate for non-small cell lung cancer (NSCLC). It uses a plasmid DNA-based vector to deliver the TUSC2 gene (a tumor suppressor gene) directly to cancer cells, which helps re-activate the body’s natural tumor suppression pathways.
  • Genprex: Reqorsa: A gene therapy candidate for non-small cell lung cancer. It uses a plasmid DNA-based vector to deliver the TUSC2 gene (a tumor suppressor gene) directly to cancer cells, which helps re-activate the body’s natural tumor suppression pathways.

Viral Vectors in Cell and Gene Therapy Overview

Lentiviruses and retroviruses are commonly used in cell and gene therapy research and production as vectors for delivering therapeutic genes into target cells. They belong to the family of retroviruses, but lentiviruses are a subset that can infect both dividing and non-dividing cells, making them particularly versatile. Learn about the differences in lentiviral vs retroviral vectors.

Lentivirus in Cell and Gene Therapy

  • Gene Delivery to Non-Dividing Cells: Unlike most retroviruses, lentiviruses can infect non-dividing cells such as neurons, muscle cells, and liver cells. This makes them highly useful for delivering therapeutic genes to a wide range of tissues in vivo.
  • Long-Term Gene Expression: Lentiviruses integrate their genetic material into the host cell’s genome, allowing for stable, long-term expression of therapeutic genes. This is essential for treatments that require sustained gene expression, such as in chronic diseases or for producing therapeutic proteins continuously.
  • Ex-Vivo Gene Therapy: Lentiviral vectors are often used in ex-vivo gene therapy. In this approach, patient cells (e.g., hematopoietic stem cells or T-cells) are removed, genetically modified using lentiviruses to introduce therapeutic genes, and then returned to the patient. This is the method used in CAR-T cell therapy, where T-cells are modified to express chimeric antigen receptors to attack cancer cells.

Retrovirus in Cell and Gene Therapy

  • Gene Delivery to Dividing Cells: Retroviruses can integrate their genetic material into the DNA of dividing cells, making them useful in cases where the target cells are rapidly proliferating, such as in some cancers or certain types of stem cell therapies.
  • Ex-Vivo Gene Therapy for Hematopoietic Stem Cells: Retroviruses are commonly used in ex-vivo gene therapy approaches where hematopoietic stem cells are removed from the patient, modified with a therapeutic gene, and then transplanted back into the patient. These stem cells can then give rise to genetically corrected blood cells, offering potential treatments for blood disorders.
  • Oncolytic Retrovirus Therapy: Retroviruses are also being engineered to target and kill cancer cells directly. This approach, called oncolytic viral therapy, involves using retroviruses that preferentially infect and destroy tumor cells, while sparing normal cells.

Viral Vector Cell and Gene Therapy Applications

Lentivirus Cell and Gene Therapy Applications

  • CAR-T Cell Therapy: Lentiviral vectors are used to modify T-cells in patients with certain types of cancers (e.g., leukemia, lymphoma) to target and kill tumor cells.
  • Treatment of Genetic Diseases: Lentivirus-mediated gene therapy is used to treat genetic disorders like β-thalassemia and sickle cell anemia by delivering functional copies of defective genes into hematopoietic stem cells.
  • Neurodegenerative Disease Research: Lentiviruses are also used in research on neurodegenerative diseases such as Parkinson’s and Huntington’s disease. They can deliver therapeutic genes into neurons, offering potential treatments for these conditions.

Retrovirus Cell and Gene Therapy Applications

  • X-linked Severe Combined Immunodeficiency (SCID-X1): One of the early successes of retrovirus mediated gene therapy was in treating SCID-X1, a rare immune disorder. By using retroviruses to deliver a functional copy of the gene responsible for SCID, researchers successfully restored immune function in patients.
  • Gene Therapy for Cancer: Retroviruses have been used to introduce therapeutic genes into cancer cells, including suicide genes that make cancer cells more susceptible to chemotherapy or other treatments.

Lentivirus Based Cell and Gene Therapy Companies and Therapeutic Candidates

  • bluebird bio:
    • Zynteglo: An approved gene therapy for beta-thalassemia, a rare genetic blood disorder. Lentiviral vectors are used to deliver a functional copy of the β-globin gene into hematopoietic stem cells.
    • Skysona: A lentiviral gene therapy for cerebral adrenoleukodystrophy (CALD), a neurodegenerative disorder. It uses lentivirus to deliver a functional gene into the patient’s HSCs.
  • Kite Pharma:
    • Yescarta: A CAR-T cell therapy for treating large B-cell lymphoma. Lentiviral vectors are used to modify the patient’s T-cells to express chimeric antigen receptors targeting cancer cells.
  • Orchard Therapeutics:
    • Libmeldy: Approved gene therapy for metachromatic leukodystrophy (MLD). It uses lentiviral vectors to deliver a functional copy of the ARSA gene into HSCs.
  • Novartis:
    • Kymriah: A CAR-T cell therapy for treating B-cell acute lymphoblastic leukemia (ALL) and large B-cell lymphoma. It uses lentiviral vectors to engineer the patient’s T-cells.

Retrovirus Based Cell and Gene Therapy Companies and Therapeutic Candidates

  • GSK:
    • Strimvelis: A retroviral gene therapy for adenosine deaminase severe combined immunodeficiency (ADA-SCID). Retroviral vectors are used to insert a functioning ADA gene into the patient’s hematopoietic stem cells.
  • Rocket Pharmaceuticals:
    • RP-L201: A retroviral gene therapy for leukocyte adhesion deficiency-I (LAD-I), using a gamma-retroviral vector to insert a functioning gene into the patient’s stem cells.
  • Mustang Bio:
    • MB-107: Retrovirus-based gene therapy for X-linked severe combined immunodeficiency (SCID-X1), also known as “bubble boy syndrome.” It uses a retroviral vector to insert a corrected version of the IL2RG gene into hematopoietic stem cells.

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