Gene Therapy II

Question 1

Thymidine Kinase Cell-Based Immunotherapy

Answer 1

Donor T-cells are transfected with HSV-TK to be given alongside bone marrow transplant
If the T-cells are alloreactive, they are weeded out with ganciclovir
Remaining cells are a boost to the immune system while the bone marrow is still generating T-cell precursors

Question 2

Cytotoxic T-cells (Tc-cells)

Answer 2

Activated by APCs
Kill target cells via perforin and granzyme
Can improve this process

Question 3

CAR-T manufacture

Answer 3

Take WBC’s from patient
Send to mantufacture, where beaded Abs activate the T-cells
Use retrovirus to reprogram T-cells to express CARs
Now they’re CAR-T’s, so multiply them EX vivo
Pt meanwhile recieves chemo that depletes immune system
Now insert CAR-T’s into pt

Question 4

Good features of Mesenchymal Stem Cells

Answer 4

Can hone to site of injury
Anti-inflammatory Action
Hypoimmunogenic - can be used off-the-shelf

Question 5

Potential uses of Mesenchymal Stem Cells

Answer 5

They can be transfected with -

Antiangiogenic genes
Genes for cytokine production
miRNAs for gene regulation
or use them as drug vehicles due to their honing ability

Question 6

CRISPR-Cas9

Answer 6

Cas9 is an enzyme that cuts two strands of DNA at a specific location in the genome
It needs a guideRNA that is a pre-designed sequence that ‘guides’ Cas9 to the right part of the genome

Question 7

CRISPR-Cas9 IN VIVO insertion

Answer 7

Use a gene vector containing Cas9, guideRNA and the DNA to be inserted

Question 8

CRISPR-Cas9 EX VIVO insertion

Answer 8

1. IMMUNOTHERAPY Remove T-cells, CRISPR edit them and reinsert them