TREATING CYSTIC FIBROSIS

Introduction

4 ways
Physiotherapy Diet and enzymes Drug therapies Transplant surgery Infertility treatments.

PHYSIOTHERAPY
Remove as much mucus from the lungs as possible Easier to breathe enough oxygen Reduces risk of lung infection Done couple times a day depending on the severity of CF New device changes the pressure in the airways, help people to clear their mucus w/o vigorous physiotherapy E.g.: Flutter the positive expiratory pressure (PEP) and thAIRapy bronchial drainage system.

Images

DIET AND ENZYMES

Thick mucus block pancreatic duct digestion and absorption of food is bad Overcome CF patients take enzymes replace the missing pancreatic enzymes food can be digested Carefully balanced diet eat more than normal people need more high energy food, high fat, high carbohydrate foods, adults must take twice the amount of proteins.

DRUG THERAPIES
Usually

take many medication cocktail

Antibiotics - tobramycin solution for inhalation Vaccines Mucolytic - Pulmozyme to thin mucus so

people can cough it out easier. Asthma drugs salbutamol and steroids DNAase enzymes Insulin

TRANSPLANT SURGERY

LUNG cannot function properly heart also affected only way is through a transplant of lung and heart New organs not affected by CF other parts will still be affected After transplant person takes IMMUNOSUPPRESSANT drugs for whole life avoid body reacting against the new tissue and rejecting it. Suppresses immune system makes it harder for the body to fight infections However most people receiving transplant usually do well

INFERTILITY TREATMENTS
Women

having CF have increasingly had babies Need to use IVF some reproductive system is not badly affected. Men techniques taking sperm from the testes and using it fertilize eggs in vitro embryos formed returned to the mothers uterus to develop normally.

GENE THERAPY

INTRODUCTION
CF

Treatments aims to remove symptoms entirely / cure Hope to replace the recessive allele with a healthy allele = GENE THERAPY GENE THERAPY involves taking a copy of the healthy gene and finding an effective way of getting it into the cells that need it to produce the correct protein.

COPYING THE HEALTHY GENE

Copying

and inserting genes genetic engineering / genetic modification = first stage of gene therapy Use special enzymes restriction endonucleases cut healthy DNA strands into specific sites. Enzymes cut DNA easier to handle each endonucleases specific restricted sites on DNA thats how the get the name!

COPYING THE HEALTHY GENE

Artificial

copies of the healthy gene made mRNA transrcibed use to produce correct DNA sequence reversing the transcription process using enzyme reverse transcriptase DNA made from this known as complementary DNA (cDNA) Since we know CFTR base sequence make these base pairs in labs.

VECTORS
Next

stage attach isolated gene to another piece of DNA VECTOR PLASMIDS circular strands of DNA found in bacteria used as vectors Plasmids combine with the cells DNA forms RECOMBINANT DNA Other vectors are needed to carry new DNA to human cells CF tested with harmless viruses and liposomes

 Viruses

infect lung epithelial cells insert viral DNA (including the added gene) into cells DNA. Liposome fuse cell membrane pass through the membrane deliver the new DNA to the cytoplasm Viruses are better to transmit the DNA but causes immune reaction in some people research focuses on non-viral not so effective Specific marker gene added easier to see when the insertion of the correct DNA has been successful

 Once

healthy gene inside the lungs the genes should transcribed and translate into producing NORMAL , ACTIVE CFTR PROTEINS relieving the symptoms BUT! : practice has shown that it is not easy to do so.

GENE THERAPY IN CYSTIC FIBROSIS

GENE THERAPY IN CF
Mutated gene chromosome 7 isolated 1989 By 1990 normal gene copied and added to cells affected by cystic fibrosis in lab proven effective Begin human studies used both virus and liposomes limited success not enough to relieve symptoms of the disease

 1999

techniques improved - 25% of chloride ions movements were restored but sodium did not seem to be moving properly. Problem epithelial cells were continuously shed and replaced new cells made by the patients body. It only contained the faulty gene new DNA that was inserted was lost relieve only lasted for a short period of time OTHE DIFFICULTIES: .

1. EXTRACELLULAR BARRIERS
Many

barrier to overcome new DNA vector reaches epithelial cells ? because CF blocks airways difficult for spray to penetrate the lungs, surface is coated with thick sticky mucus. Ways dilute mucus-Mucolytic/DNAase trying different routes get vectors to lungs through the blood.

2. INTRACELLULAR BARRIERS
Vector

inside the cell to get DNA to the correct place Proved to be very difficult especially when liposomes are used as vectors 1/1000 plasmids that enter the liposome gets to the nucleus to be transcribed Research focuses on modifying the vectors to get the new genes into the nuclei more effectively Viruses gives best results!

3. KEEPING THE GENE EXPRESSION GOING

Once

inside the nucleus transcribed problem it does not work for long 2 weeks at best! Research looking at ways to extend this and make it permanent by using different vectors Repair the faulty allele on the original DNA itself if stem cells could be used lead to permanent cure.

CONCLUSION
The

hope for gene therapy is that people will not need treatment after initial medication Quality of life and expectancy improved with no respiratory complications Big financial saving without lifelong treatments Treat effects of the gut from the disease in the same way It is hoped that by 2009 2010 that human trial will be underway again, possibly on a larger scale than before!

THE END THANK YOU !