Lecture - Genetics (Recombinant Proteins + Gene Therapy)

Question 1

Haemophilia: fill this in

Answer 1

Question 2

Haemophilia A genetics:

1. What deficiency is it in?
2. What are the three classifications?
3. What is the inheritance pattern (dominant, recessive etc)
4. In apparently isolated cases (1/3 of cases), how much percent of the cases have mothers who are carriers? Often the mutation arises how?
5. There are polymorphisms that don’t cause the disease (normal variants), but with mild/moderate/severe, what sorta mutations are we seeing?
6. Whereabouts is the gene for factor 8 (haemophilia A)? Is it big or small? What sorta hetrogeniety are we looking at?
7. Why are inversions common in terms of mutations of this gene?
8. Where is factor 8 synthesised? Why is it processed through the ER?
9. What is factor 8 activated by?
10. How is factor 8 involved in the coagulation cascade?
11. What is the half life of factor 8? What does this mean in terms of treatment?

Answer 2

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Question 3

What are the three products that have been used to increase survival and reduce complicatons?

Answer 3

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Question 4

Concentrate:

1. How much does it aim to increase the bleeding?
2. The protein is purified in a complex with _____ and concentrated from large amounts of ______ donated blood using biochemical techniques
3. What’s the life expectancy like? What does it mean it’s given prophylactically?
4. Why were there concerns about blood safety after 1980s? (talk about how the procedures may not completely inactivate….)
5. Is the processing expensive? But several products….
6. What is supply of concentrate dependent on?
7. What is the name of the concentrate named in NZ (it’s made in Aus by NZ donors)
8. Contains both factor 8 and?

Factor 8 inhibitors:

1. What may the immune system do against the foreign protein?
2. How many pateints would this affect?
3. How do you treat this?

Answer 4

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Question 5

Recombinant process:

1. What cells do they use and why?
2. With the cells they use - where are they grown, how are they used, what don’t they have etc? So they are grown and then what happens? P_____
3. However, expression levels are still ____, and the tissue culture methid and purification is _____
4. First generation product required the addition of what? Second and third generation products don’t require it and instrad use what? Why is this important?
5. So like, treatment is expensive but what’ll raise the price even more?
6. So before we talked about developing inhibitors for the concentrate, how about now for the recombinant protein? Is the inhibitor rate similar for the two F8 products?

Question 6

Comparing concentrate vs recombinant:

1. What’s the cost of trearment like? But then what about safety?
2. What about supplies of recombinant?

3 What’s expensive in this recombinant process?

Question 7

1. What’re the three limitations of protein therapy?
2. Gene therapy is a one-hit treatment. But what three things do we need for it?

Answer 7

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Question 8

Gene therapy:

1. What is it?
   - to replace…
   - to inhibit….
   - to kill…..
2. What’re the two main methods?
   - administered ex vivo or in vivo - meaning?
   - expression is regulated for what? What two expressions?

Answer 8

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Question 9

Engineered or recombinant viral vectors

1. What are the three major types?
2. Is any vector perfect?
3. Viruses for gene transfer - what is replaced with promotor and gene of interest? So how does this promotor tie in with overexpression?
4. What is SCID? How is it treated now vs bubble boy?
5. Stem cells sources for gene therapy - what is direct delivery vs cell-based delivery?

Answer 9

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Question 10

Gene therapy progress in haemophilia

1. Is it a good gene therapy target? Why or why not?
2. What’ve the clinical trials been like for haemophilia?

Question 11

Batten’s disease:

1. Why is this a difficult disease to treat with gene therapy?
2. Protein therapy and gene therapy are both difficult - why?
3. Protein (enzyme) therapy for CLN2
   - what’s it got to do with BBB?
   - needs direct infusion - how frequent?
   - sucessfully delaying disease onset but what?
   - how do we treat it now?
4. WIth gene therapy - single infusion into what to get it to brain?
   - should be an improvement on enzyme ____

Answer 11

2. Need to get it through BBB
3. Answer the questions
4. Idk, it jst said it’s more difficult