医学遗传学 19遗传疾病的治疗eng v-精选文档.ppt

Medical Genetics 1. What is gene therapy? Genes, which are carried on chromosomes, are the basic physical and functional units of heredity. Genes are specific sequences of bases that encode instructions on how to make proteins. Medical Genetics Although genes get a lot of attention, its the proteins that perform most life functions and even make up the majority of cellular structures. Medical Genetics When genes are altered so that the encoded proteins are unable to carry out their normal functions, genetic disorders can result. Medical Genetics Gene therapy is a technique for correcting defective genes responsible for disease development. Medical Genetics Researchers may use one of several approaches for correcting faulty genes: 1. A normal gene may be inserted into a nonspecific location within the genome to replace a nonfunctional gene. This approach is most common. 2. An abnormal gene could be swapped for a normal gene through homologous recombination. 3. The abnormal gene could be repaired through selective reverse mutation, which returns the gene to its normal function. 4. The regulation (the degree to which a gene is turned on or off) of a particular gene could be altered. Medical Genetics 2. How does gene therapy work? In most gene therapy studies, a “normal“ gene is inserted into the genome to replace an “abnormal,“ disease-causing gene. Medical Genetics A carrier molecule called a vector must be used to deliver the therapeutic gene to the patient's target cells. Currently, the most common vector is a virus that has been genetically altered to carry normal human DNA. Medical Genetics Viruses have evolved a way of encapsulating and delivering their genes to human cells in a pathogenic manner. Medical Genetics Scientists have tried to take advantage of this capability and manipulate the virus genome to remove disease-causing genes and insert therapeutic genes. Medical Genetics Target cells such as the patient's liver or lung cells are infected with the viral vector. The vector then unloads its genetic material containing the therapeutic human gene into the target cell. Medical Genetics The generation of a functional protein product from the therapeutic gene restores the target cell to a normal state. Medical Genetics To reverse disease caused by genetic damage, researchers isolate normal DNA and package it into a vector, a molecular delivery truck usually made from a disabled virus. Doctors then infect a target cell usually from a tissue affected by the illness, such as liver or lung cellswith the vector. The vector unloads its DNA cargo, which then begins producing the missing protein and restores the cell to normal. Medical Genetics A. Some of the different types of viruses used as gene therapy vectors (1) Retroviruses A class of viruses that can create double-stranded DNA copies of their RNA genomes. These copies of its genome can be integrated into the chromosomes of host cells. Human immunodeficiency virus (HIV) is a retrovirus. Medical Genetics (2)Adenoviruses A class of viruses with double-stranded DNA genomes that cause respiratory, intestinal, and eye infections in humans. The virus that causes the common cold is an adenovirus. (3)Adeno-associated viruses A class of small, single-stranded DNA viruses that can insert their genetic material at a specific site on chromosome 19. (4)Herpes simplex viruses A class of double-stranded DNA viruses that infect a particular cell type, neurons. Herpes simplex virus type 1 is a common human pathogen that causes cold sores. Medical Genetics Medical Genetics Medical Genetics Medical Genetics Besides virus-mediated gene- delivery systems, there are several nonviral options for gene delivery. The simplest method is the direct introduction of therapeutic DNA into target cells. This approach is limited in its application because it can be used only with certain tissues and requires large amounts of DNA. Medical Genetics Another nonviral approach involves the creation of an artificial lipid sphere with an aqueous core. This liposome, which carries the therapeutic DNA, is capable of passing the DNA through the target cell's membrane. Medical Genetics Therapeutic DNA also can get inside target cells by chemically linking the DNA to a molecule that will bind to special cell receptors. Medical Genetics Once bound to these receptors, the therapeutic DNA constructs are engulfed by the cell membrane and passed into the interior of the target cell. This delivery system tends to be less effective than other options. Medical Genetics Researchers also are experimenting with introducing a 47th artificial human chromosome into target cells. This chromosome would exist autonomously alongside the standard 46 -not affecting their workings or causing any mutations. Medical Genetics It would be a large vector capable of carrying substantial amounts of genetic code, and scientists anticipate that, because of its construction and autonomy, the body's immune systems would not attack it. A problem with this potential method is the difficulty in delivering such a large molecule to the nucleus of a target cell. Medical Genetics 3. The current status of gene therapy research Current gene therapy is experimental and has not proven very successful in clinical trials. Medical Genetics Little progress has been made since the first gene therapy clinical trial began in 1990. In 1999, gene therapy suffered a major setback with the death of 18-year-old Jesse Gelsinger. Medical Genetics Jesse was participating in a gene therapy trial for ornithine transcarboxylase deficiency (OTCD). He died from multiple organ failures 4 days after starting the treatment. His death is believed to have been triggered by a severe immune response to the adenovirus carrier. Medical Genetics Another major blow came in January 2003, when the FDA (USA) placed a temporary halt on all gene therapy trials using retroviral vectors in blood stem cells. Medical Genetics FDA (USA) took this action after it learned that a second child treated in a French gene therapy trial had developed a leukemia-like condition. Medical Genetics Both this child and another who had developed a similar condition in August 2002 had been successfully treated by gene therapy for X-linked severe combined immunodeficiency disease (X-SCID), also known as “bubble baby syndrome.“ Medical Genetics FDA's Biological Response Modifiers Advisory Committee (BRMAC) met at the end of February 2003 to discuss possible measures that could allow a number of retroviral gene therapy trials for treatment of life-threatening diseases to proceed with appropriate safeguards. FDA has yet to make a decision based on the discussions and advice of the BRMAC meeting. Medical Genetics 4. Factors kept gene therapy effective A. Short-lived nature of gene therapy Before gene therapy can become a permanent cure for any condition, the therapeutic DNA introduced into target cells must remain functional and the cells containing the therapeutic DNA must be long-lived and stable. Medical Genetics Problems with integrating therapeutic DNA into the genome and the rapidly dividing nature of many cells prevent gene therapy from achieving any long-term benefits. Patients will have to undergo multiple rounds of gene therapy. Medical Genetics B. Immune response Anytime a foreign object is introduced into human tissues, the immune system is designed to attack the invader. Medical Genetics The risk of stimulating the immune system in a way that reduces gene therapy effectiveness is always a potential risk. Medical Genetics Furthermore, the immune system's enhanced response to invaders it has seen before makes it difficult for gene therapy to be repeated in patients. Medical Genetics C. Problems with viral vectors Viruses, while the carrier of choice in most gene therapy studies, present a variety of potential problems to the patient -toxicity, immune and inflammatory responses, and gene control and targeting issues. Medical Genetics In addition, there is always the fear that the viral vector, once inside the patient, may recover its ability to cause disease. Medical Genetics D. Multigene disorders Conditions or disorders that arise from mutations in a single gene are the best candidates for gene therapy. Medical Genetics Unfortunately, some the most commonly occurring disorders, such as heart disease, high blood pressure, Alzheimer's disease, arthritis, and diabetes, are caused by the combined effects of variations in many genes. Medical Genetics Multigene or multifactorial disorders such as these would be especially difficult to treat effectively using gene therapy. Medical Genetics 5. Some recent developments University of California, Los Angeles, research team gets genes into the brain using liposomes coated in a polymer call polyethylene glycol (PEG). The transfer of genes into the brain is a significant achievement because viral vectors are too big to get across the “blood-brain barrier.“ This method has potential for treating Parkinson's disease. Medical Genetics RNA interference or gene silencing may be a new way to treat Huntington's. Short pieces of double- stranded RNA are used by cells to degrade RNA of a particular sequence. If a siRNA is designed to match the RNA copied from a faulty gene, then the abnormal protein product of that gene will not be produced. Medical Genetics Medical Genetics New gene therapy approach repairs errors in messenger RNA derived from defective genes. Technique has potential to treat the blood disorder thalassaemia, cystic fibrosis, and some cancers. Medical Genetics Gene therapy for treating children with X-SCID (sever combined immunodeficiency) or the “bubble boy“ disease is stopped in France when the treatment causes leukemia in one of the patients. Medical Genetics Researchers at Case Western Reserve University and Copernicus Therapeutics are able to create tiny liposomes 25 nanometers across that can carry therapeutic DNA through pores in the nuclear membrane. Medical Genetics 6. Some of the ethical considerations What is normal and what is a disability or disorder, and who decides? Are disabilities diseases? Do they need to be cured or prevented? Does searching for a cure demean the lives of individuals presently affected by disabilities? Medical Genetics Who will have access to these therapies? Who will pay for their use? Is somatic gene therapy more or less ethical than germline gene therapy (which is done in egg and sperm cells and prevents the trait from being passed on to further generations)?