Gene therapy and stem cells are discussed. [1] Gene therapy involves delivering genetic material into cells for therapeutic purposes using vectors. It aims to replace defective genes, enhance gene expression, or suppress harmful genes. [2] Stem cells are undifferentiated cells that can develop into specialized cell types and have self-renewal abilities. There are embryonic, adult tissue specific, and induced pluripotent stem cells. Gene therapy and stem cells offer potential treatments for diseases like cancer, genetic disorders, and tissue regeneration. However, safety and delivery challenges remain.
Gene therapy is an experimental technique that uses genes to treat or prevent disease. The slides explain what is gene tharapy? Types of gene therapy. http://www.wesrch.com/
Gene therapy is an experimental technique that uses genes to treat or prevent disease. The slides explain what is gene tharapy? Types of gene therapy. http://www.wesrch.com/
Gene Therapy, Somatic cell gene therapy, germ line gene therapy, classical gene therapy, non-classical gene therapy, targets of gene therapy, barriers of gene therapy, ex vivo gene therapy, in vivo gene therapy, vectors for gene delivery, antisense therapy
Gene therapy is the process of inserting therapeutic genes into cells to prevent or cure wide range of diseases. The newly introduced genes will encode proteins and correct the deficiencies that occur in genetic diseases. Gene therapy primarily involves genetic manipulations in animals or humans to correct a disease, and keep organism in good health. It is a technique for correcting defective genes responsible for disease and development.
In this slide, You will get to learn abut Gene Therapy and different types of gene therapy. Various method of Gene Therapy and Advantage & Disadvantage and Recent advances in Gene Therapy.
INTRODUCTION
DNA VACCINES
GENE THERAPY
TIME LINE OF DEVELOPING GENE THERAPY
GENE THERAPY STRATEGIES
TECHNOLOGY OF CLASSICAL GENE THERAPY
PRINCIPLES OF GENE TRANSFER
VECTORS
VIRAL VECTORS
NON-VIRAL VECTORS
APPLICATIONS OF GENE THERAPY
ETHICAL IMPLICATIONS
THE FUTURE
CONCLUSION
REFERENCES
Nucleic Acid Based Therapeutic Delivery System.pptxPrachi Pandey
The delivery of nucleic acid molecules into cells to alter physiological functions at the genetic level is a powerful approach to treat a wide range of inherited and acquired disorder.
This technique has been a common research tool in laboratory for decades to study gene functions.
The therapeutic potential of this approach was not fully realized due to lack of reliable and practical methods to transfer and express recombinant DNA in mammalian cells.
Gene Therapy, Somatic cell gene therapy, germ line gene therapy, classical gene therapy, non-classical gene therapy, targets of gene therapy, barriers of gene therapy, ex vivo gene therapy, in vivo gene therapy, vectors for gene delivery, antisense therapy
Gene therapy is the process of inserting therapeutic genes into cells to prevent or cure wide range of diseases. The newly introduced genes will encode proteins and correct the deficiencies that occur in genetic diseases. Gene therapy primarily involves genetic manipulations in animals or humans to correct a disease, and keep organism in good health. It is a technique for correcting defective genes responsible for disease and development.
In this slide, You will get to learn abut Gene Therapy and different types of gene therapy. Various method of Gene Therapy and Advantage & Disadvantage and Recent advances in Gene Therapy.
INTRODUCTION
DNA VACCINES
GENE THERAPY
TIME LINE OF DEVELOPING GENE THERAPY
GENE THERAPY STRATEGIES
TECHNOLOGY OF CLASSICAL GENE THERAPY
PRINCIPLES OF GENE TRANSFER
VECTORS
VIRAL VECTORS
NON-VIRAL VECTORS
APPLICATIONS OF GENE THERAPY
ETHICAL IMPLICATIONS
THE FUTURE
CONCLUSION
REFERENCES
Nucleic Acid Based Therapeutic Delivery System.pptxPrachi Pandey
The delivery of nucleic acid molecules into cells to alter physiological functions at the genetic level is a powerful approach to treat a wide range of inherited and acquired disorder.
This technique has been a common research tool in laboratory for decades to study gene functions.
The therapeutic potential of this approach was not fully realized due to lack of reliable and practical methods to transfer and express recombinant DNA in mammalian cells.
NUCLEIC ACID BASED THERAPEUTIC DELIVERY SYSTEM by pramesh..pptxPRAMESHPANWAR1
Name of the title: Nucleic Acid-Based Therapeutic Delivery System.
It includes information about nucleic acid, gene therapy, and its type, a method to deliver the desired DNA, i.e., vectors and their types, with proper examples and diagrams, and how these things help in delivering a nucleic acid-based therapeutic drug delivery system.
Nucleic Acid Based Therapeutic Delivery System.pptxRAHUL PAL
Therapeutic nucleic acids (TNAs) are nucleic acids themselves or closely related compounds used to treat disease. Although various types of TNAs exist, they share a common mechanism of action that is mediated by sequence‐specific recognition of endogenous nucleic acids through Watson–Crick base pairing 7.
What are the advantages of nucleic acid based therapeutics?
The major advantage of nucleic acid-based therapeutics lies in the fact that they can be used to accurately target a tumor or tissue, then have a specific therapeutic protein, biologic, or immune engager expressed only at the site of interest.
Gene therapy refers to the insertion of genetic material to correct a genetic defect.
In gene therapy, a "normal" gene is inserted into the genome to replace an "abnormal," disease-causing gene
Prix Galien International 2024 Forum ProgramLevi Shapiro
June 20, 2024, Prix Galien International and Jerusalem Ethics Forum in ROME. Detailed agenda including panels:
- ADVANCES IN CARDIOLOGY: A NEW PARADIGM IS COMING
- WOMEN’S HEALTH: FERTILITY PRESERVATION
- WHAT’S NEW IN THE TREATMENT OF INFECTIOUS,
ONCOLOGICAL AND INFLAMMATORY SKIN DISEASES?
- ARTIFICIAL INTELLIGENCE AND ETHICS
- GENE THERAPY
- BEYOND BORDERS: GLOBAL INITIATIVES FOR DEMOCRATIZING LIFE SCIENCE TECHNOLOGIES AND PROMOTING ACCESS TO HEALTHCARE
- ETHICAL CHALLENGES IN LIFE SCIENCES
- Prix Galien International Awards Ceremony
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Ve...kevinkariuki227
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Verified Chapters 1 - 19, Complete Newest Version.pdf
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Verified Chapters 1 - 19, Complete Newest Version.pdf
Ozempic: Preoperative Management of Patients on GLP-1 Receptor Agonists Saeid Safari
Preoperative Management of Patients on GLP-1 Receptor Agonists like Ozempic and Semiglutide
ASA GUIDELINE
NYSORA Guideline
2 Case Reports of Gastric Ultrasound
Tom Selleck Health: A Comprehensive Look at the Iconic Actor’s Wellness Journeygreendigital
Tom Selleck, an enduring figure in Hollywood. has captivated audiences for decades with his rugged charm, iconic moustache. and memorable roles in television and film. From his breakout role as Thomas Magnum in Magnum P.I. to his current portrayal of Frank Reagan in Blue Bloods. Selleck's career has spanned over 50 years. But beyond his professional achievements. fans have often been curious about Tom Selleck Health. especially as he has aged in the public eye.
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Introduction
Many have been interested in Tom Selleck health. not only because of his enduring presence on screen but also because of the challenges. and lifestyle choices he has faced and made over the years. This article delves into the various aspects of Tom Selleck health. exploring his fitness regimen, diet, mental health. and the challenges he has encountered as he ages. We'll look at how he maintains his well-being. the health issues he has faced, and his approach to ageing .
Early Life and Career
Childhood and Athletic Beginnings
Tom Selleck was born on January 29, 1945, in Detroit, Michigan, and grew up in Sherman Oaks, California. From an early age, he was involved in sports, particularly basketball. which played a significant role in his physical development. His athletic pursuits continued into college. where he attended the University of Southern California (USC) on a basketball scholarship. This early involvement in sports laid a strong foundation for his physical health and disciplined lifestyle.
Transition to Acting
Selleck's transition from an athlete to an actor came with its physical demands. His first significant role in "Magnum P.I." required him to perform various stunts and maintain a fit appearance. This role, which he played from 1980 to 1988. necessitated a rigorous fitness routine to meet the show's demands. setting the stage for his long-term commitment to health and wellness.
Fitness Regimen
Workout Routine
Tom Selleck health and fitness regimen has evolved. adapting to his changing roles and age. During his "Magnum, P.I." days. Selleck's workouts were intense and focused on building and maintaining muscle mass. His routine included weightlifting, cardiovascular exercises. and specific training for the stunts he performed on the show.
Selleck adjusted his fitness routine as he aged to suit his body's needs. Today, his workouts focus on maintaining flexibility, strength, and cardiovascular health. He incorporates low-impact exercises such as swimming, walking, and light weightlifting. This balanced approach helps him stay fit without putting undue strain on his joints and muscles.
Importance of Flexibility and Mobility
In recent years, Selleck has emphasized the importance of flexibility and mobility in his fitness regimen. Understanding the natural decline in muscle mass and joint flexibility with age. he includes stretching and yoga in his routine. These practices help prevent injuries, improve posture, and maintain mobilit
New Directions in Targeted Therapeutic Approaches for Older Adults With Mantl...i3 Health
i3 Health is pleased to make the speaker slides from this activity available for use as a non-accredited self-study or teaching resource.
This slide deck presented by Dr. Kami Maddocks, Professor-Clinical in the Division of Hematology and
Associate Division Director for Ambulatory Operations
The Ohio State University Comprehensive Cancer Center, will provide insight into new directions in targeted therapeutic approaches for older adults with mantle cell lymphoma.
STATEMENT OF NEED
Mantle cell lymphoma (MCL) is a rare, aggressive B-cell non-Hodgkin lymphoma (NHL) accounting for 5% to 7% of all lymphomas. Its prognosis ranges from indolent disease that does not require treatment for years to very aggressive disease, which is associated with poor survival (Silkenstedt et al, 2021). Typically, MCL is diagnosed at advanced stage and in older patients who cannot tolerate intensive therapy (NCCN, 2022). Although recent advances have slightly increased remission rates, recurrence and relapse remain very common, leading to a median overall survival between 3 and 6 years (LLS, 2021). Though there are several effective options, progress is still needed towards establishing an accepted frontline approach for MCL (Castellino et al, 2022). Treatment selection and management of MCL are complicated by the heterogeneity of prognosis, advanced age and comorbidities of patients, and lack of an established standard approach for treatment, making it vital that clinicians be familiar with the latest research and advances in this area. In this activity chaired by Michael Wang, MD, Professor in the Department of Lymphoma & Myeloma at MD Anderson Cancer Center, expert faculty will discuss prognostic factors informing treatment, the promising results of recent trials in new therapeutic approaches, and the implications of treatment resistance in therapeutic selection for MCL.
Target Audience
Hematology/oncology fellows, attending faculty, and other health care professionals involved in the treatment of patients with mantle cell lymphoma (MCL).
Learning Objectives
1.) Identify clinical and biological prognostic factors that can guide treatment decision making for older adults with MCL
2.) Evaluate emerging data on targeted therapeutic approaches for treatment-naive and relapsed/refractory MCL and their applicability to older adults
3.) Assess mechanisms of resistance to targeted therapies for MCL and their implications for treatment selection
micro teaching on communication m.sc nursing.pdfAnurag Sharma
Microteaching is a unique model of practice teaching. It is a viable instrument for the. desired change in the teaching behavior or the behavior potential which, in specified types of real. classroom situations, tends to facilitate the achievement of specified types of objectives.
Title: Sense of Taste
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the structure and function of taste buds.
Describe the relationship between the taste threshold and taste index of common substances.
Explain the chemical basis and signal transduction of taste perception for each type of primary taste sensation.
Recognize different abnormalities of taste perception and their causes.
Key Topics:
Significance of Taste Sensation:
Differentiation between pleasant and harmful food
Influence on behavior
Selection of food based on metabolic needs
Receptors of Taste:
Taste buds on the tongue
Influence of sense of smell, texture of food, and pain stimulation (e.g., by pepper)
Primary and Secondary Taste Sensations:
Primary taste sensations: Sweet, Sour, Salty, Bitter, Umami
Chemical basis and signal transduction mechanisms for each taste
Taste Threshold and Index:
Taste threshold values for Sweet (sucrose), Salty (NaCl), Sour (HCl), and Bitter (Quinine)
Taste index relationship: Inversely proportional to taste threshold
Taste Blindness:
Inability to taste certain substances, particularly thiourea compounds
Example: Phenylthiocarbamide
Structure and Function of Taste Buds:
Composition: Epithelial cells, Sustentacular/Supporting cells, Taste cells, Basal cells
Features: Taste pores, Taste hairs/microvilli, and Taste nerve fibers
Location of Taste Buds:
Found in papillae of the tongue (Fungiform, Circumvallate, Foliate)
Also present on the palate, tonsillar pillars, epiglottis, and proximal esophagus
Mechanism of Taste Stimulation:
Interaction of taste substances with receptors on microvilli
Signal transduction pathways for Umami, Sweet, Bitter, Sour, and Salty tastes
Taste Sensitivity and Adaptation:
Decrease in sensitivity with age
Rapid adaptation of taste sensation
Role of Saliva in Taste:
Dissolution of tastants to reach receptors
Washing away the stimulus
Taste Preferences and Aversions:
Mechanisms behind taste preference and aversion
Influence of receptors and neural pathways
Impact of Sensory Nerve Damage:
Degeneration of taste buds if the sensory nerve fiber is cut
Abnormalities of Taste Detection:
Conditions: Ageusia, Hypogeusia, Dysgeusia (parageusia)
Causes: Nerve damage, neurological disorders, infections, poor oral hygiene, adverse drug effects, deficiencies, aging, tobacco use, altered neurotransmitter levels
Neurotransmitters and Taste Threshold:
Effects of serotonin (5-HT) and norepinephrine (NE) on taste sensitivity
Supertasters:
25% of the population with heightened sensitivity to taste, especially bitterness
Increased number of fungiform papillae
Couples presenting to the infertility clinic- Do they really have infertility...Sujoy Dasgupta
Dr Sujoy Dasgupta presented the study on "Couples presenting to the infertility clinic- Do they really have infertility? – The unexplored stories of non-consummation" in the 13th Congress of the Asia Pacific Initiative on Reproduction (ASPIRE 2024) at Manila on 24 May, 2024.
These lecture slides, by Dr Sidra Arshad, offer a quick overview of physiological basis of a normal electrocardiogram.
Learning objectives:
1. Define an electrocardiogram (ECG) and electrocardiography
2. Describe how dipoles generated by the heart produce the waveforms of the ECG
3. Describe the components of a normal electrocardiogram of a typical bipolar leads (limb II)
4. Differentiate between intervals and segments
5. Enlist some common indications for obtaining an ECG
Study Resources:
1. Chapter 11, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 9, Human Physiology - From Cells to Systems, Lauralee Sherwood, 9th edition
3. Chapter 29, Ganong’s Review of Medical Physiology, 26th edition
4. Electrocardiogram, StatPearls - https://www.ncbi.nlm.nih.gov/books/NBK549803/
5. ECG in Medical Practice by ABM Abdullah, 4th edition
6. ECG Basics, http://www.nataliescasebook.com/tag/e-c-g-basics
Lung Cancer: Artificial Intelligence, Synergetics, Complex System Analysis, S...Oleg Kshivets
RESULTS: Overall life span (LS) was 2252.1±1742.5 days and cumulative 5-year survival (5YS) reached 73.2%, 10 years – 64.8%, 20 years – 42.5%. 513 LCP lived more than 5 years (LS=3124.6±1525.6 days), 148 LCP – more than 10 years (LS=5054.4±1504.1 days).199 LCP died because of LC (LS=562.7±374.5 days). 5YS of LCP after bi/lobectomies was significantly superior in comparison with LCP after pneumonectomies (78.1% vs.63.7%, P=0.00001 by log-rank test). AT significantly improved 5YS (66.3% vs. 34.8%) (P=0.00000 by log-rank test) only for LCP with N1-2. Cox modeling displayed that 5YS of LCP significantly depended on: phase transition (PT) early-invasive LC in terms of synergetics, PT N0—N12, cell ratio factors (ratio between cancer cells- CC and blood cells subpopulations), G1-3, histology, glucose, AT, blood cell circuit, prothrombin index, heparin tolerance, recalcification time (P=0.000-0.038). Neural networks, genetic algorithm selection and bootstrap simulation revealed relationships between 5YS and PT early-invasive LC (rank=1), PT N0—N12 (rank=2), thrombocytes/CC (3), erythrocytes/CC (4), eosinophils/CC (5), healthy cells/CC (6), lymphocytes/CC (7), segmented neutrophils/CC (8), stick neutrophils/CC (9), monocytes/CC (10); leucocytes/CC (11). Correct prediction of 5YS was 100% by neural networks computing (area under ROC curve=1.0; error=0.0).
CONCLUSIONS: 5YS of LCP after radical procedures significantly depended on: 1) PT early-invasive cancer; 2) PT N0--N12; 3) cell ratio factors; 4) blood cell circuit; 5) biochemical factors; 6) hemostasis system; 7) AT; 8) LC characteristics; 9) LC cell dynamics; 10) surgery type: lobectomy/pneumonectomy; 11) anthropometric data. Optimal diagnosis and treatment strategies for LC are: 1) screening and early detection of LC; 2) availability of experienced thoracic surgeons because of complexity of radical procedures; 3) aggressive en block surgery and adequate lymph node dissection for completeness; 4) precise prediction; 5) adjuvant chemoimmunoradiotherapy for LCP with unfavorable prognosis.
2. What is gene Therapy ?
Means of delivering exogenous genetic material
for therapeutic purposes into the host cell target
using vectors.
Basic Principle: Intrinsic expressions of certain
genes in the body tissues can be modified to
treat disease.
3. Done by ..
Replacing the defective gene with a functional version
Introduction of functioning ADA gene into the bone marrow of children with SCID
using a viral vector - reconstituting the patient’s immune system
Enhancing the baseline expression level of a gene
Gene for TNF Introduced into TILs ex-vivo and when transfused into patient
preferentially migrates to residual tumor providing the therapeutic TNF.
Suppressing the expression of genes that may contribute to the
pathologic process.
Introduction of RNAi by any means into a patient selectively turn off the gene
in disease(Cancer),blocking the mRNA translation.
4. HISTORY..
1960’s : The concepts of Gene Therapy was introduced.
1972 : Friedman and Roblin authored a paper in Science titled "Gene
therapy for human genetic disease.”
1984: A retrovirus vector system was designed that could efficiently insert
foreign genes into mammalian chromosomes.
1990: The first FDA approved gene therapy in the US took place on four
year old Ashanti DeSilva for a genetic defect that left her with ADA-SCID, a
severe immune system deficiency.
5. 1992: Doctor Claudio Bordignon performed the first procedure of gene
therapy using hematopoietic stem cells as vectors to deliver genes
intended to correct hereditary diseases.
1999: Death of Jesse Gelsinger in a gene-therapy experiment
2003 : a research team inserted genes into the brain for the first time
using liposomes.
2006 : Successful use of gene therapy to treat two adult patients for X-
linked chronic granulomatous disease.
2007: first gene therapy trial for inherited retinal disease
6. 2010 : an 18 year old male patient in France with beta-thalassemia major had been
successfully treated.
2011: Medical community accepted that it can cure HIV as in 2008, Gero Hutter has
cured a man from HIV using gene therapy (repeated haematopoietic stem cell
transplantation)
July 2012, European commission approved alipogene tiparvovec(Glybera) ,an AAV
viral vector harbouring human lipoprotein lipase for familial lipoprotein lipase
deficiency disease.First commercial available gene therapy technology in Europe.
Research is still ongoing and the number of diseases that has been treated
successfully by gene therapy increases.
Most notable being gene therapy for ADA-SCID,B-Thalassemia.Sickle cell
disease,CFTR
7. As of 2012, some 1800 human clinical gene therapy trials (including phase I to
IV) exist world wide.The disease process involved can be broadely divided into
four groups (descending order)
Cancer
Monogenetic disease(CF,Haemophilia B)
Cardiovascular disease
Infectious disease (HIV)
8. Delivery
In the field of gene therapy,development of vehicle for the introduction of genetic
material into selected target cells has been a major focus.
Major barriers
Targeting: only cells that require gene
Binding and internalization (Transfection)
Cellular trafficking to the nucleus(Transduction):Need to avoid endosomal
degradation
Nuclear expression: Quantity and stability of the gene expression need to be
determined.
Different methods varies in ability in overcoming these barriers.
9. Methods
Vehicles of gene delivery are known as vectors
Ideal vector
TARGET the right cells
INTEGRATE the gene in the cells.
ACTIVATE the gene.
AVOID harmful side effects.
But, no universal vector exists.
10. Two general gene delivery methods(vectors) can be mentioned.
Viral and
Non-viral vectors
11. Non-viral vector
Most extensively studied method.
Includes
1) Plasmid mediated transfer of genes
Gene can be effectively introduced into muscle and thyroid simply by injecting
DNA into tissues.
Lipofection and gene gun methods- enhanced uptake of DNA in other tissues.
Advantages: Simplicity, Safe (do not alter cell’s genome),minimal immune response
Obstacle: Efficiency
12. 2 ) Direct delivery of RNA for gene silencing
IV administration of siRNA targeting VEGF gene has shown to reduce
significant tumor volume and intratumoral VEGF levels.
Lipofection: Liposomes significantly improves both the duration and level of
gene expression compared with non vector delivery methods.
13. Viral mediated gene transfer:
Till date majority of the research has been focussed on developing
methods of using viruses as vectors.
Viruses introduce their genetic material into the host cell as part of their
replication cycle
Involves construction of synthetic virus particles bearing the properties:
1.Lack pathogenic function
2.Incapable of replication
3.Contain a therapeutic gene within the viral genome and
4.Can deliver the genes to the cells by the process of infection.
14. Viral mediated gene transfer:
Advantages:
Target specific types of cells. So, they're very good at targeting and entering cells.
Can be modified so that they can't replicate and destroy cells.
Certain virus can permanently integrate their genes into chrosomes of the
cell-Permanent gene therapy
Can be introduced for both the dividing and non dividing cells
Disadvantage:
Immune reaction,Toxicity or may cause disease inside.
15. Common viruses that have been used for human gene
therapy includes:
Adenovirus
Adeno-associated virus and
Retrovirus/lentivirus
16. Retrovirus:
Original prototypes
Useful properties:
Permanent gene therapy
Modifications can be made – Enhanced safety features
Limitations:
Only integrate into actively dividing cells and efficiency is relatively low
Solution: lentiviral vectors
Most serious: Difficulty in achieving stable,regulated gene expression despite
permanent genomic integration.
17. Complications:
Serious complications have been reported in clinical trials for X-linked SCID
Cases of T-cell leukemia have been reported
Solution: Suicide gene inclusion in vector genome that will initiate cell death in
overproliferating cells.
18. Adeno Virus:
Powerful and effective vehicle for gene transfer.
Features:
Unlike retrovirus the virus remain episomal, so the virus do not integrate
genes into target cells chromosome
Advantage
Infecting both dividing and non dividing cells
Both in- vivo and ex vivo with high level of efficiency.
19. Adeno-Associated virus:
Permanently integrates into target cell chromosome but unlike retrovirus can stably
infect both dividing and nondividing cells for prolonged periods.
Safe options:
Apparent lack of pathogenecity
Do not induce insertional mutagenesis or an innate immune response.
Disadvantage:
Purified from the potentially cytotoxic,wild type helper virus.
Limited capacity(4.8kb)
20. Strategies for administering gene
therapy:
Ex-vivo
Cells isolated from surgical biopsy are
grown in culture,modified(genes inserted
using typically retroviral vector) and the
cells are reimplanted in the body via
autologous transplantation
Creates a population of cells within the
body that permanently express a
therapeutic function
Cells are treated outside.
E.g.1st Gene Therapy done in a SCID
patient.
In-vivo
DNA or viral vectors ,predominantly
adenoviral vectors are administrated directly
to patients.
Intent can be single treatment for certain
tumors,administered intermittently in
response to acute disease or given long term
to establish steady state level of therapeutic
gene product.
E.g. Adenovirus mediated replacement of
defective CFTR gene in CF.
22. Gene Therapy: Types
1) Somatic vs germ cell gene therapy based on possible target cells and
1) Permanent vs Temporary gene therapy.
23. Somatic cell Gene Therapy
Genes transferred to somatic cells
(Bonemarrow cells,Blood cells,Skin cells)
Not inherited to later generations
At present all researches directed to
correct genetic defects in somatic cells.
Embryonic cell Gene Therapy
Genes transferred to germ cells
( Eggs and sperms)
Inherited to future generation
For safety, ethical and technical
reasons, it is not being attempted at
present
Genetic manipulation of germ cells prohibited under
existing recombinant DNA guidelines.
24. Permanent Gene Therapy
More desirable when the therapy involves
significant surgical procedures or substantial
risks( like organ resection)
Requires tight control and regulation of
transferred genetic material and
Not feasible with present techniques.
(growth hormone deficiency,juvenile diabetes)
Temporary Gene Therapy
Beneficial for diseases like cancer, arthritis, CF
and disorders that require surgery.
Demand is limited, one time gene expression or
repeat gene transfer needed over period of
time could prove effective
Noninvasive routes like IM.IV or even aerosol
administration allow steady-state gene product
levels
25. Why Gene Therapy??
New Therapeutic approach:
Novel approach to disease that are not satisfactorily managed using
conventional pharmacologic or surgical intervention.
E.g. Gives alternative to allogenic transplantation of bone marrow,solid
organs,or individual cells.
Site specific gene expression:
Therapeutic products can be released from specific cell types in precise
locations within the body.
26. Hurdles..
Short lived nature: need multiple treatment
Immune response:
Viral vectors: Toxicity,gene control and targeting tissues
Multigene disorders: Like Heart disease,Diabetes. Complicates gene therapy
Insertional Mutagenesis:T cell leukemia during trial for X-SCID.
Cost: Alipogene tiparvovec or glybera,famed as “million dollar drug” cost 31 million
Euro as of 2015 per trearment.
Deaths: Three deaths have been reported
1st Jesse Gelsinger 1999 because of immune rejection response.
2nd X-SCID patient died of leukemia in 2003.
3rd in 2007, a rheumatoid arthritis patient died from an infection
27. Why Gene Therapy??
Improved efficacy and safety:
Establishes the expression of normal human proteins in a directed
therapeutic fashion within the body maintaining efficacy and safety.
Improved route of administration and compliance:
Provides continuous endogenous expression of natural protein products
and requires less frequent administration (of gene) ranging from one time
to weekly or monthy treatment depending upon disease target and
transfer strategy.
28. Why Gene Therapy??
Preventive medicine and reduction in health care costs:
Gene Therapy can establish continuous release of a therapeutic product
with one time treatment or infrequent dosing
Its use for prevention is more practical and affordable than conventional
therapies.
Atherosclerosis,cancer,diabetes,infections or degenerative diorders.
29. Application of Gene Therapy in
OTOLARYNGOLOGY
Relatively new.
Inherited Disease:
Includes sinus disease in CF, hearing loss in Usher,Pendred or Alport syndrome and
goiter in congenital hypothyroidism.
First clinical trial for CF involved introduction of normal CFTR gene into nasal mucosa
using adenoviral vectors that provided foundation for the studies in which the CFTR
gene would be replaced throughout the respiratoty tract using viral or DNA vectors.
30. Application of Gene Therapy in
OTOLARYNGOLOGY
Head and Neck ONCOLOGY
Melanoma,Squamous cell carcinoma
Approaches:
Immune Modulation Approaches:
1.Genetic modification of Tumor-Infiltrating Lymphocytes
Gene therapy involves transfer of the gene for IL-2 directly into a patients tumor
resulting in local formation of tumor specific cytolytic TIL cells.Il-2 increases the
immunogenesity of cancer cells and suppress the tumor growth.
31. 2. Direct In Vivo stimulation of a Antitumor Immune Response:
Introduction of genes for various cytokines directly into tumor cells to increase the
natural immune response to tumor specific antigens.
In head and neck cancer animal model,use of adenoviral mediated delivery of the
cytokine IL-2 in combination with a cytotoxic gene –synergistic effect on tumor
regression.
Note:In a phase I clinical trial patients received TNFerade(2nd generation adenoviral vector with
TNFa gene inserted) in addition to hysroxyurea,5-FU,and reirradiation-response achieved 83% of
patients,although survival remained very poor in study cohort.
32. Suicide gene Therapy:
Infecting tumor cells by direct injection of a gene with retrovirus that encodes
enzymes like thymidine kinase - making them vulnerable and susceptible to
chemotherapy.
33. Gene augmentation strategies:
Modifying oncogenes and tumor suppressor genes:
TP53 and CDKN2A mutation - Head and NECK cancer.
Growing evidence from phase I and II trials, significant benefit from
mediated TP53 gene therapy
Use of novel dominant negative Adenovirus vector that encodes a mutant RAD50
gene, causes downregulation of MRN expression leading to sensitization of
SCC cells to cisplatin with increased apoptosis.
34. Gene suppression strategies:
Gene silencing:
Selective turning off or silencing genes to prevent development or progression of disease.
Study says,silencing of EGFR gene (HNSCC) using antisense(plasmid mediated) technique
causes greater cytotoxic effect than conventional therapy.
shRNA use in silencing cyclin D1 gene markedly sensitizes (HNSCC) cells to cisplatin in vitro
0n Targeting E6 and E7 oncogenes(upregulation of P53 and PRb) – causes cytotoxic effect
HPV related oropharyngeal carcinoma.
35.
36. Inhibiting angiogenesis:
Introduction of the gene that inhibit angiogenesis in vicinity of a tumor-
regression.
E10A-Adenoviral vector with gene that encodes endostatin - Inhibit
angiogenesis
37. Oncolytic viruses:
Early studies- viral vectors were replication incompetent
Now,genetically engineered to selectively infect and replicate in targeted tumor
cells that have inherent genetic defects such as loss of TP53 gene expression
leading to tumor kill.
Onyx-015(adenovirus): First replication selective viral vector on human trial.
Key-deletion of P53 binding protein gene that causes selective replication in
tumor cells lacking P53 expression causing lysis of target cells and spread to nearby
cells. More benefit on combination therapy .
38. Plastic and Reconstructive Surgery
Principle:
Expression of growth regulating factors to enhance repair or regeneration of
damaged tissues and to enhance local proliferation to fill surgical defects.
Reconstrcutive tissue flaps and wound healing
Gene encoding angiogenesis factor(bFGF),cytokines and growth factors(TGF-a,IGF-
1-2,PDGF) promotes tissue repair and regeneration.
39. Plastic and Reconstructive Surgery
Skin grafting :
Feasibility of cultivating epidermal cells ex-vivo and subsequently engrafting the cells
Nerve Injury:
viral vectors(Adenovirus,lentivirus,herpes simplex) can be used to prevent death of
motoneurons and enhance axonal regeneration
Optimizes bone growth:
In case of removal of bone and soft tissue loss in extensive HNSCC
Repair and regeneration of irradiated tissues by enhancing viability and vascularity
40. Laryngology
IGF-1 with its neutropic and myotropic effects holds potential for
optimizing function after acute laryngeal injury or for augmenting surgical
re innervation of the paralysed larynx.
41. Otology AND Neurotology
Experimental stage.
Studies have demonstrated that adenoviral delivery of gene Atoh1 gene to
inner ear of guinea pigs result in production and innervation of inner ear
cells.
43. Stem cells
Unspecialized & undifferentiated cells that are able to develop into any
specialized type of cells.
Features:
Self renewel
Potency
44. Types:
Embryonic Stem cells
Somatic or adult tissue specific Stem cells
Induced Pluripotent stem cells
45. Adult tissue specific Stem cells:
Immature cell that is capable of dividing indefinitely both to renew itself and
produce cells maturing for tissue function.
Found in most normal and malignancy tissues.
46. Multipotent:
Produce many types of cells that in one family e.g. Hematoid stem cells (make
types of the blood & lymph cells
Oligopotent:
Can produce Few types of cells•e.g. lymphoid stem cells (make many types of
lymphoid cells like T, B, Nkcells
Unipotent:
can produce one type of cells e.g. Muscle stem cell
48. Embryonic Stem Cells
Totipotent stem cells(a.k.a.
omnipotent)
Stem cells can differentiate into
embryonic and extraembryonic cell
types,like the Amnion, Yolk sac, Chorion,
placenta, Umbilical Cord
The Zygote & the cells produced by the
first few divisions of the fertilized egg
are also totipotent (Zygote divides into
2, 4, 8, 16 cells ”morula”)
Toti: means ‘whole’as they make the whole
organism
Pluripotent Stem cells
Stem cells are the descendants of totipotent
cells and can differentiate into nearly all
cells (except the extraembryonic cell )
The inner cell mass, Bilamenlar &
Trilamenlar germ disc
The differentiate into more than 200 types
of cells
Pluri: means ‘Several’
49. Induced Pluripotent stem cells (iPSCs)
Induced expression of stem cell genes such as Oct4,Sox2,cMyc and Klf4 in
differentiated cells results in acquisition of stem cell properties similar to
those of Embryonic Stem cells.
Potential clinical use for replacement of tissues that are genetically
defective or have been lost to damage or disease.
50. Application of stem cells:
Otolaryngology
The ear and nose:
Engineered cartilage grafts generated from cartilage stem cells in biopsies
from nasal septum can be used for reconstruction.
Cochlea damage or degeneration:
Loss of inner hair cells is currently irreversible but certain stem cells
procedure has been reported to generate hair cells and associated nerve.
51. Trachea:
Small defect-Autologous cartilage graft
Extensive tissue loss(malignancy):Tissue engineering can be used to generate
tracheal tissues from autologous stem cells.
Larynx and vocal cords:
Regenerative medicine has the potential to enhance VF recovery beyond that
associated with conventional treatment.
Other tissues:
Muscle derived Multipotent Stem Cells have been shown to improve outcomes
facial and RLN injuries.
Most of DNA is not internalized,even if it is internalized endosomal degradation of nearly all remaining DNA.Since Plasmid uptake is not receptor mediated targeting to a specific cell is a major obstacle.Expression is also transient since plasmid is lost with cell division.
Gene gun:
uses electrical current to project microscopic gold beads coated with plasmid DNA into target cells.
Prior studies: Adoptive transfer of TIL cells coupled with administration of IL-2 causes significant tumor regression in some patiensts with Malignant Melanoma.
MRN complex is critical for repair of genomic damage and survival even with high dose chemotherapeutic agents.Use of novel dominant negative adenovirus vector that encodes a mutant RAD50 gene,causes downregulation of MRN expression and disruption of function leading to sensitization of human SCC cells to cisplatin with increased apoptosis.
TP53 and CDKN2A tumor suppressor genes mutation - Head and NECK cancer.