ENGR / BRAE 213 – Bioengineering Fundamentals
Biotechnology
Case Study- Duchenne Muscular Dystrophy (DMD)
Dystrophin gene
Case Study- Duchenne Muscular Dystrophy (DMD)M
uscle fiber
Dystrophin gene
Case Study- Duchenne Muscular Dystrophy (DMD)M
uscle fiber
Dystrophin gene
Case Study- Duchenne Muscular Dystrophy (DMD)
Case Study- Duchenne Muscular Dystrophy (DMD)
Genomics
• How do we know patient has mutation?– Sequencing– PCR
Genomics- Background
“expressed gene”
DNA makes up genome
Mutations (single nucleotide polymorphism)
• Different nucleotides in DNA/RNA can lead to different amino acids in protein
Person 1
Person 2
Genomics Technology- PCR
• Polymerase Chain Reaction – amplify DNA
Genomics Technology- PCR
• http://highered.mcgraw-hill.com/olc/dl/120078/micro15.swf
• Produce specificity with primers– 1 primer set for normal DNA– 1 primer set to recognize mutated region
Genomics Technology- PCR
Genomics Technology- Sequencing
• Step 1 – amplify target sequence– dideoxynucleotides (ddNTPs) stop replication by
polymerase
• Step 2- electrophoresis of products
Genomics Technology- Sequencing
Genomics Technology- Sequencing
Genomics
• Sequencing explained:– http://youtu.be/bEFLBf5WEtc
• Using PCR or sequencing- can identify mutations in the patients’ genome and use that information to inform treatment
Pharmacological Therapy
Symptom
Anti-inflammatorysteroidInflammation
Gene Therapy
• Bioengineering as practical application of biological science– Use genes themselves to directly treat
patients
Fiber Death
InflammationFiber Regeneration
Cause
Gene Therapy
• Naked DNA
• Virus containing DNA
• Liposome containing DNA– Lipid bilayer
• Naked DNA + simple, safe, efficient uptake into cell
– poor incorporation into genome, tissue specific
• Virus containing DNA+ efficient uptake & incorporation, specific
– immune rejection, inflammation
• Liposome containing DNA+ efficient uptake
– non-specific, poor incorporation intogenome
Gene Therapy
FIRST U.S. TRIAL OF DMD GENE THERAPY UNDER WAYCOLUMBUS, Ohio, March 29, 2006 — The first U.S. human gene therapy trial directed at Duchenne muscular dystrophy (DMD) was launched yesterday at Columbus (Ohio) Children’s Hospital, the Muscular Dystrophy Association (MDA), Children’s Hospital, and Asklepios Biopharmaceutical Inc. (AskBio) announced today.
DMD research study halted?
Activity
Which delivery method would be preferable for treat the intestine?
A. Naked DNA
B. Virus containing DNA
C. Liposome containing DNA
Which delivery method would be preferable for treat the
intestine?
A. Naked DNA
B. Virus containing DNA
C. Liposome containing DNA
Activity
Which delivery method would be preferable for delivery to the retina?
A. Naked DNA
B. Virus containing DNA
C. Liposome containing DNA
Which delivery method would be preferable for delivery to the
retina?
A. Naked DNA
B. Virus containing DNA
C. Liposome containing DNA
Protein Therapy
• Fiber regeneration ≈ fiber enlargement
Cloned myostatin gene to obtain sequence
Used sequence information to delete myostatin gene from genome in mice
Fiber Death
InflammationFiber Regeneration
Limited
by myostatin
Fiber Death
InflammationFiber Regeneration
Limited
by myostatin
Protein Therapy
• Myostatin antibody to block protein from functioning
Str
engt
h
Protein Therapy
• Myostatin antibody to block protein from functioning
Question:
If you were a DMD patient, what would be your preferred form of treatment?
A. Dystrophin gene therapy
B. Myostatin antibody therapy
C. Prednisone (anti-inflammatory)
D. Physical therapy