Gene Therapy Basics Quiz
45 Questions
0 Views

Choose a study mode

Play Quiz
Study Flashcards
Spaced Repetition
Chat to lesson

Podcast

Play an AI-generated podcast conversation about this lesson

Questions and Answers

What term describes the introduction of foreign DNA into another cell via a viral vector?

  • Transduction (correct)
  • Gene therapy
  • Titer
  • Infection

    • Monogenic disorders are caused by multiple genes.

    False (B)

    What is the main purpose of using a carrier vector in gene therapy?

    To deliver the therapeutic gene to target cells.

    The concentration of viral particles is commonly expressed in units called ______.

    <p>Infectious Units per ml (IU/ml)</p> Signup and view all the answers

    Match the following genetic conditions with their descriptions:

    <p>Monogenic = Caused by one gene Polygenic = Requires many genes to manifest Inherited disorders = Genetic conditions passed from parent to offspring Environmental factors = External conditions affecting disease occurrence</p> Signup and view all the answers

    What is the primary goal of Gene Therapy?

    <p>To treat diseases by altering genes (C)</p> Signup and view all the answers

    Gene Therapy only involves the removal of genes from the individual's cells.

    <p>False (B)</p> Signup and view all the answers

    What was the first gene therapy clinical study conducted for?

    <p>A genetic defect causing an immune system deficiency (ADA-SCID)</p> Signup and view all the answers

    Gene Therapy is the insertion, alteration, or removal of genes within an individual's __________ to treat disease.

    <p>cells and tissues</p> Signup and view all the answers

    Match the following terms related to Gene Therapy with their definitions:

    <p>Transformation = Transfer of naked DNA into bacteria Transfection = Transfer of naked DNA into cells RNAi = Suppression of gene expression Genome Editing = Modification of genes at specific locations</p> Signup and view all the answers

    Which of the following methods is involved in Gene Therapy?

    <p>Alteration, addition, or suppression of genes (C)</p> Signup and view all the answers

    There is a consensus on the definition of Gene Therapy with no ongoing controversy.

    <p>False (B)</p> Signup and view all the answers

    Name one ethical concern related to Gene Therapy.

    <p>Potential long-term effects on recipients or genetic modifications in germline cells.</p> Signup and view all the answers

    What type of molecule is a plasmid?

    <p>Circular double-stranded DNA (D)</p> Signup and view all the answers

    Bacterial transformation allows bacteria to take up DNA from their own chromosomes.

    <p>False (B)</p> Signup and view all the answers

    Who was awarded the Nobel Prize in 1958 for his work relating to bacteria?

    <p>Joshua Lederberg</p> Signup and view all the answers

    Adenovirus is a class of virus that typically causes __________ infections.

    <p>respiratory</p> Signup and view all the answers

    Match the following characteristics with their respective terms:

    <p>Plasmid = Circular double-stranded DNA molecule Bacterial transformation = Uptake of foreign DNA Adenovirus = Non-integrating virus Gene delivery = Method of introducing genetic material into cells</p> Signup and view all the answers

    What is a major limitation of plasmid-mediated gene delivery?

    <p>Low gene transfer efficiency (A)</p> Signup and view all the answers

    Viral vectors can integrate into the genome of host cells.

    <p>True (A)</p> Signup and view all the answers

    What is the primary advantage of using plasmids as gene carriers?

    <p>They can replicate independently in bacteria.</p> Signup and view all the answers

    What is one major feature of retroviral vectors?

    <p>They integrate their genome into the host cell genome. (C)</p> Signup and view all the answers

    Insertional mutagenesis is a potential risk associated with gene therapy using integrating vectors.

    <p>True (A)</p> Signup and view all the answers

    What is ADA-SCID?

    <p>A rare disorder caused by the absence of the enzyme adenosine deaminase, impairing immune system development.</p> Signup and view all the answers

    Children with ADA-SCID are often diagnosed with SCID within the first _____ months of life.

    <p>6</p> Signup and view all the answers

    Match the following conditions with their corresponding characteristics:

    <p>X linked SCID = Inherited immune deficiency Chronic Granulomatous disease = Impaired ability to kill certain bacteria Wiskott Aldrich Syndrome = Combined immune deficiency with eczema ADA-SCID = Deficiency in adenosine deaminase</p> Signup and view all the answers

    Which of the following is a treatment option for ADA-SCID?

    <p>Matched bone marrow transplant (C)</p> Signup and view all the answers

    Lentiviral vectors do not integrate into the host cell genome.

    <p>False (B)</p> Signup and view all the answers

    What are the main symptoms of ADA deficiency?

    <p>Pneumonia, chronic diarrhea, widespread skin rashes, and developmental delays.</p> Signup and view all the answers

    What is a notable characteristic of lentiviral vectors in terms of cell division?

    <p>They can transduce host cells without requiring active cell division. (A)</p> Signup and view all the answers

    Lentiviral vectors are more likely to activate adjacent oncogenes compared to gammaretroviral vectors.

    <p>False (B)</p> Signup and view all the answers

    What type of gene therapy caused a severe immune reaction, leading to the death of Jesse Gelsinger?

    <p>Adenoviral vector therapy</p> Signup and view all the answers

    The CRISPR/Cas9 system has significantly impacted __________ and biotechnology.

    <p>scientific research</p> Signup and view all the answers

    Match the following incidents with their corresponding gene therapies:

    <p>Jesse Gelsinger = Adenoviral vector therapy First gene therapy trials = 1st gen. retroviral vectors XLMTM trial = AAV treatment T cell leukemia development = X-SCID treatment</p> Signup and view all the answers

    What was a primary safety concern in the initial gene therapy trials for X-SCID?

    <p>Unexpected development of T cell leukemia (D)</p> Signup and view all the answers

    Self-Inactivating (SIN) designs aim to decrease the risk of genotoxicity by retaining strong enhancer elements.

    <p>False (B)</p> Signup and view all the answers

    How many children died in the gene therapy trial for X-linked myotubular myopathy (XLMTM) due to liver failure?

    <p>Four</p> Signup and view all the answers

    What is the primary application of CRISPR/Cas9 technology mentioned in the content?

    <p>Treating genetic diseases (C)</p> Signup and view all the answers

    CRISPR/Cas9 was recognized with a Nobel Prize in 2020.

    <p>True (A)</p> Signup and view all the answers

    What are the two genetic conditions mentioned that CRISPR/Cas9 may help to treat?

    <p>Sickle cell disease and thalassemia</p> Signup and view all the answers

    CRISPR/Cas9 technology is primarily used for ___ medicine.

    <p>genetic</p> Signup and view all the answers

    Match the following terms related to CRISPR/Cas9 with their descriptions:

    <p>CRISPR = A genetic engineering tool Cas9 = An enzyme that cuts DNA CAR-T cell therapy = A treatment method for cancer Gene therapy = Introducing or altering genetic material</p> Signup and view all the answers

    Which of the following is a feature of CAR-T cell therapy?

    <p>It genetically modifies a patient’s own T cells. (A)</p> Signup and view all the answers

    CRISPR/Cas9 can only be used for treating inherited diseases.

    <p>False (B)</p> Signup and view all the answers

    What is the role of hemoglobin in the human body?

    <p>Carrying oxygen within red blood cells</p> Signup and view all the answers

    Study Notes

    Gene Therapies Overview

    • Gene therapy is the insertion, alteration, or removal of genes within an individual's cells and tissues to treat disease.
    • There is still debate about the exact definition and its scope.
    • Methods include suppression with RNAi, antisense oligonucleotides, increased/restored gene expression, and gene modification using genome editing.

    Who is Axel Hyrenius Wittsten?

    • Researcher in Synthetic Immunology
    • Has a PhD from Lund University, Sweden
    • Currently a Postdoctoral researcher at the University of California, San Francisco (UCSF), USA
    • Works on gene therapies and biopharmaceuticals.
    • His email address is [email protected].

    Lecture Objectives

    • MAIN:
      • Understand the broad concept of gene therapy.
      • Learn about the methods of gene delivery and correction.
      • Identify advantages, disadvantages, and limitations of different delivery systems.
    • Supplementary:
      • Explore the potential of gene therapy in real-world clinical applications.
      • Examine the capabilities and ethical challenges of gene therapy.
      • Cultivate an enthusiasm for the field.

    Lecture Content

    • PART I: General introduction and classification of gene therapy
    • PART II: Detailed descriptions of gene delivery systems and their applications.
    • PART III: Case studies illustrating the transition from proof-of-principle to clinical trials.

    Defining Gene Therapy

    •  Aimed at curing diseases by modifying genes within an organism's cells and tissues.
    • This involves inserting, altering, or removing genes to treat diseases, currently an evolving field.
    • Different methods are used, including suppression via RNAi and antisense oligonucleotides, restoring or increasing gene expression, and gene modification techniques.

    Categories of Gene Therapy

    • Type of Disease: Genetic or Acquired.
    • Delivery Vehicle: Integrating or Non-integrating.
    • Type of Administration: In vivo or Ex vivo.

    Origin and Early Clinical Study

    • Gene therapy proposals emerged in the 1970s.
    • The first gene therapy clinical trial in 1990 targeted ADA-SCID in a four-year-old girl at NIH.

    Gene Delivery Terminology

    • Transformation: Transferring naked DNA into bacteria.
    • Transfection: Transferring naked DNA into cells.
    • Infection: Introducing a wild-type virus into cells.
    • Transduction: Using a viral vector to transfer DNA into another cell.
    • Titer: Measurement of viral particles' concentration (Infectious Units/ml).

    Genetic Background of Diseases

    • Genetic Factors:
      • Monogenic disorders (single gene mutations).
      • Polygenic disorders (multiple gene mutations).
    • Environmental Factors: Infectious diseases or combined with genetic factors.
      • Common diseases: cancers, chronic diseases.

    Faulty Genetics: Types

    • Loss-of-function: Normal gene function is disrupted and absent.
    • Gain-of-function: The gene produces unintended or abnormal protein products.

    Basis of Gene Therapy

    • Gene therapy uses a "carrier vector" to deliver therapeutic genes.
    • Vectors, mostly viral (albeit modified), carry normal human DNA into target cells.
    • Gene delivery results in normal functioning of the affected cells.

    Advanced Gene Delivery Methodologies

    • Viral vectors:
      • Advantages and disadvantages of different types.
    • Non-viral vectors:
      • Nanoparticle-based approaches for safe and efficient delivery.
      • Ex. liposomes, solid lipid nanoparticles, niosomes, polymeric nanoparticles, lipopeptide nanoparticles.
    • Methods of delivery:
      • In vivo: introduce vectors directly into the patient's body,
      • Ex vivo: isolate, alter cells, and re-introduce them into the body

    Ex Vivo Vs. In Vivo

    • Ex vivo: Modify cells outside the body then return to target cells that have been reprocessed.
    • In vivo: Vectors are introduced directly into the patient.

    Ex Vivo: Self or Non-self (Cellular Source)

    • Autologous: Use of the patient's own cells.
    • Allogeneic: Use of cells from a donor.

    Ex Vivo: Advantages of Both Strategies

    • Autologous: Patient-specific, no rejection by immune system, no risk of graft-vs-host disease, and repeated doses possible
    • Allogeneic: Larger-scale manufacturing possible, reduced donor variability, more reproducible manufacturing, and rapid availability

    Ex Vivo: Challenges of Both Strategies

    • Autologous: High costs for manufacturing and quality testing, varying starting material variability
    • Allogeneic: Risk of graft-vs-host disease, cost increase due to minimized risk of rejection, and the risk of donor cells' immunogenicity.

    Delivery Vehicle: Non-viral

    • Liposome-based nanoparticles, Solid-lipid nanoparticles, and Niosomes.
    • Polymer-based nanoparticles, and Lipopeptide-based nanoparticles.

    Non-viral Example: Hereditary variant transthyretin amyloidosis (ATTRv)

    • A rare genetic disorder.
    • Mutant TTR protein misfolds and forms aggregates.
    • TTR aggregates build up in various organs and tissues.
    • Patisiran (liposomal siRNA) targets TTR, reducing accumulation and improving clinical symptoms.

    Non-viral Example: COVID-19 mRNA vaccines

    • Use of nucleoside-modified mRNA.
    • mRNA triggers the immune response, inducing production of specific antibodies.

    Plasmids

    • Circular double-stranded DNA molecules.
    • Can self-replicate inside bacterial cells, independent of their chromosome.
    • Used as gene carriers.
    • Efficient method is bacterial transformation.

    Plasmid-Mediated Gene Delivery

    • Challenges: Low transfer efficiency, only transient expression, and difficulties in delivering to primary cells.
    • Recombinant viral vectors (retroviral, lentiviral, adenoviral, AAV) offer superior methods for inserting genes.

    Delivery Vehicle: Viral

    • Adenovirus, Adeno-associated virus (AAV), Retrovirus, and Lentivirus are vectors for delivery.

    • Features: sizes, genome types/sizes, packaging capacity, transduction efficiency, integration ability, and immunogenicity

    Viral Vectors (Non-integrating viruses)

    • Adenovirus: Causes common colds, its DNA doesn’t integrate into host cells.
    • Adeno-Associated Virus (AAV): Small, single-stranded DNA virus, non-pathogenic, and does not integrate into host cells.

    Viral Vectors (Integrating viruses)

    • Retroviruses, eg: Lentiviruses (eg. HIV): RNA viruses, create double-stranded DNA from their RNA genomes, capable of integrating into chromosomes.
    • Gammaretroviruses: Infect dividing cells.
    • Lentiviruses: Unique ability to infect both dividing and nondividing cells.

    Adenoviral Vectors

    • Adenoviruses are double-stranded DNA (dsDNA) viruses that do not integrate into the host cell, so gene expression is transient.
    •  High efficiency and high production capacity.
    • Risk of pre-existing immunity.

    Adeno-Associated Viral Vectors (AAV)

    • Small in size, have a relatively smaller packing capability.
    • Can transduce dividing and non-dividing cells, have low immunogenicity.
    • Transduction of cells as ds circular episomes (chromosomal integration also occurs).
    • Effective use for delivering genes into various cells and organs.

    AAVs in Clinical Applications

    • Used in a variety of clinical trials, like Hemophilia B, retinal diseases, Parkinson's disease, cystic fibrosis, and lysomal storage diseases, as well as other applications.

    Clinical Examples: Hemophilia B

    • AAV vector delivers Factor IX Padua transgene.
    • Bioengineered AAV vectors have shown significant improvement for patients.

    Viral Vectors (Integrating viruses)

    • The structure elements of retroviruses:
      • The long terminal repeats (LTRs) regulate viral transcription, acting as viral promoters.
      • The ψ sequence controls packing signals.
      • The gag proteins are structural in nature, whilst pol and env are involved in viral replication and integration and envelope protein respectively.

    Recombinant Integrating Vectors

    • Separating viral components on "packaging" and "transfer" plasmids is typical.
    • Genes to be integrated are commonly inserted into specific viral components.

    Retroviral Vectors

    • The vector's genomic material, or DNA, integrates into the host cell's genome.
    • Successful transduction of host cells involves dividing cells.
    • Potential risk of insertional mutagenesis.

    Insertional Mutagenesis

    • Insertion of viral or vector components into a gene can induce mutations, leading to disease-related issues.
    • The risk of this is a crucial aspect of retroviral vector research and its evaluation.

    Historical Overview of HSC Gene Therapy

    • The history of HSC gene therapy, demonstrating evolution of methodologies and use in clinical applications, with focus on the isolation, cloning, and cloning of viruses, and also gene transfer to murine HSCs.
    • Illustrates the journey of development; from early isolation of HSC, successful gene transfer to murine HSCs, engineering of y-retroviral vectors, advances in lentiviral vectors, improved transfer to human and NHP HSCs, and editing methods (ZFN, TALEN, CRISPR/Cas).
    • Major milestones and issues in HSC gene therapies.

    Clinical Examples: Retroviruses (Primary Immunodeficiences)

    • X-linked severe combined immunodeficiency (SCID), adenosine deaminase (ADA) SCID, Wiskott-Aldrich syndrome, and Chronic Granulomatous diseases.

    Clinical Example: ADA-SCID

    • Rare disorder characterized by mutations causing an absence of adenosine deaminase.
    • Leads to immune deficiency, usually causing death before the first year.
    • Treatment involves matched-related or unrelated bone marrow transplants; more commonly enzyme replacement therapy (ADA enzyme with PEG).
    • The early development and application of gene therapy are also highlighted, especially the use of retroviruses in early gene therapy approaches.
    • The historical perspective on HSC gene therapies provides insights into the challenges and milestones achieved in the field.

    CAR-T Cell Therapy

    • CAR-T cell therapy re-engineers a patient's T cells to target and eliminate specific cancer cells.
    • T cells are genetically altered to express artificial receptors that recognize and bind specific tumor antigens.
    • T cell receptors for binding to cancer cells are genetically introduced into T cells to deliver them to the site of the tumor.

    CAR-T Cell Production

    • Acquire T cells from the patient's blood, create CAR-T cells by inserting gene for CAR into T cells, expand the engineered T cells, infuse the engineered CAR-T cells back into the patient, and CAR T cells attack cancer cells.

    CAR-T Therapy in B Cell Malignancies

    • Used in leukemia and lymphoma cases to eliminate tumor cells expressing CD19.

    Patient "Proof-of-Principle"

    • Demonstrates the use of CAR-T therapy.
    • Shows successful treatment on children with relapsed, previously unresponsive leukemia.

    Clinical Success of CAR-T Cell Therapy

    • KYMRIAH (Tisagenlecleucel) approved for children and teens with relapsed/refractory acute lymphoblastic leukemia.
    • Initial studies demonstrated success for treatment against cancer in 27/30 patients with relapse.

    CAR-T Therapy: Major Features

    • Pros: Uses patient's own cells, eliminating graft-versus-host disease risk, and potential for lasting immunity.
    • Cons: High cost, lengthy processing time required, adverse events like cytokine release syndrome (CRS).

    Broadening CAR-T Therapy

    • Growing use of CAR-T cell therapies, expanding to various cancers.
    • Research is also focused on optimizing treatments and expanding its applicability to more patients, including the geographic expansion of these clinical trials.

    Functional Challenges in CAR-T Cell Therapy

    • Trafficking: Getting CAR T cells to the target tumor site.
    • Recognition: CAR-T cells successfully recognizing and binding to tumor targets
    • Control: Managing the activation states to limit collateral damage
    • Proliferation & Persistence: Maintaining long-lasting anti-tumor effects.

    Multi-Antigen Recognition of Tumors

    • The need for therapies that target multiple antigens rather than single ones for enhanced efficacy in tumor elimination is now clear.

    New Class of Synthetic Environmental Sensor

    • Sensor: measures the environment around cells in order to program changes.

    Developing New Clinically Relevant Circuit CAR-T Cells

    • SynNotch CAR circuits show enhanced solid tumor recognition and promote persistent antitumor activity.

    Mesothelioma

    • Heavily associated with asbestos exposure, and incidence is increasing worldwide, with long incubation periods.
    • Highly aggressive disease with limited treatment options, and often therapy resistance.

    ALPPL2 as a Novel Tumor-Specific Antigen

    • A new tumor-specific antigen.
    • High expression in mesotheliomas, seminomas, and other cancers.

    Combinatorial Targeting using ALPPL2

    • Targeting multiple antigens (ALPPL2, MCAM, HER2, or MSLN) with CAR T cells can provide enhanced tumor-specific activity.
    • Combination of different targets on the CAR-T cells may provide improved antitumor activity compared to single target CAR-T cells.

    SynNotch CAR-T Cells: Superior Efficacy in vivo

    • Superior efficacy in preclinical models for targeting mesothelioma.
    • Effectiveness also extends to other tumor types: SK-OV-3 and K562.

    ALPPL2 synNotch for other tumor types

    • Tested in cell line models, including SKOV-3 ovarian cancer cells and K562 leukemia cells, to evaluate its efficiency in cell killing.

    Next Step toward the Clinic: Modular design of Synthetic Receptors

    • The development of modular synthetic receptors allows for precise regulation of gene expression.

    Problems with SynNotch

    • Issues with the design, function, and implementation of SynNotch receptors for CAR-T cell therapy.

    New Humanized Targeting Version

    • Humanized version of ALPPL2 and HER2 targeting shows effective dual-antigen targeting in preclinical ovarian cancer models.

    Current Status: ArsenalBio

    • ArsenalBio has initiated a Phase 1 clinical trial for their ovarian cancer treatment (AB-1015) using CAR T-cells.

    Concluding Remarks

    • The rate of innovation in gene and cell therapies is outpacing the ability to translate that innovation into safe therapies. Gene editing and regulatory science advancements will be key in the future of individualized therapies.

    Three Essential Tools

    • Adeno-associated viral (AAV) vector, lentiviral vector, and gene editing complex as needed tools.

    Suggested Reading

    • Provides useful resources for further research and understanding in the context of gene therapies.

    Studying That Suits You

    Use AI to generate personalized quizzes and flashcards to suit your learning preferences.

    Quiz Team

    Related Documents

    Gene Therapies - AHW PDF

    Description

    Test your knowledge on gene therapy concepts with this engaging quiz. From the role of viral vectors to ethical concerns, explore key topics that define the field. Perfect for students looking to enhance their understanding of genetic engineering.

    More Like This

    Correcting Faulty Genes
    5 questions

    Correcting Faulty Genes

    SofterFuchsia avatar
    SofterFuchsia
    Medical Genetics: Gene Therapy Trials Overview
    38 questions

    Medical Genetics: Gene Therapy Trials Overview

    StableEpilogue avatar
    StableEpilogue
    Terapia Génica y Vacunas
    24 questions

    Terapia Génica y Vacunas

    RespectableAccordion avatar
    RespectableAccordion
    MIC 115 Lecture 13: Viral Vectors
    29 questions

    MIC 115 Lecture 13: Viral Vectors

    cookietongs avatar
    cookietongs
    Use Quizgecko on...
    Browser
    Open
    Browser
    Browser