Method and apparatus for close-field electroporation
The present invention relates to improved methods for transfecting one or more cells within a target region with an agent by electroporation. The method comprises exposing one or more cells to the agent and to a close electric field created between an anode or anode array and a cathode or cathode array in the target region for sufficient time to allow at least some of the agent to enter said one or more cells.
1. A method of transfecting one or more cells within a target region with an agent by electroporation, said method comprising using a linear electrode array comprising at least eight electrodes contiguous in an array structure provided in a single probe introduced into a target treatment region such that there will be no tissue between the electrodes due to the array structure, the electrodes of the linear array configured as a single anode array comprising four or more neighbouring electrodes and a single cathode array comprising four or more neighbouring electrodes where the distance between the cathode array and the anode array is not more than 5 mm, exposing said one or more cells to said agent and applying an electroporation pulse profile to create a close electric field focused between the anode array and cathode array in a target region within millimetre or sub-millimetre dimensional space adjacent to the array in said target region for sufficient time to cause electroporation to allow at least some of said agent to enter said one or more cells, wherein each of said anode array and said cathode array independently comprise from 4 to 8 electrodes.
2. A method of transfecting one or more cells within a target region with an agent by electroporation, said method comprising using a linear electrode array comprising eight or more electrodes contiguous in an array structure provided in a single probe introduced into a target treatment region such that there will be no tissue between the electrodes due to the array structure, the electrodes of the linear array configured as a single anode array comprising four neighbouring electrodes and a single cathode array comprising four neighbouring electrodes where the distance between the cathode array and the anode array is not more than 5 mm, exposing said one or more cells to said agent and applying an electroporation pulse profile to create a close electric field focused between the anode array and cathode array in a target region within millimetre or sub-millimetre dimensional space adjacent to the array in said target region for sufficient time to cause electroporation to allow at least some of said agent to enter said one or more cells, wherein each of said anode array and said cathode array comprises 4 electrodes.
3. The method of claim 1 , wherein the agent comprises a nucleic acid molecule which on entering said one or more cells enables said one or more cells within said target region to produce, enhance or suppress an activity within said region.
4. The method of claim 1 , wherein the agent comprises a nucleic acid molecule which on entering said one or more cells at least partially restores physiological function within said target region, one or more regions annexing said target region, or both within said target region and one or more regions annexing said target region.
5. The method of claim 1 , wherein the agent comprises a nucleic acid molecule.
6. The method of claim 5 , further characterised by any one of:
the nucleic acid molecule encodes a neurotrophic factor,
the nucleic acid molecule encodes a transcription factor which modulates the expression of a neurotrophic factor, or
the nucleic acid molecule decreases the expression of a transcription factor, wherein the transcription factor modulates the expression of a neurotrophic factor.
7. The method of claim 6 , wherein the neurotrophic factor is selected from any one of Neurotrophin-3, Neurotrophin-3 precursor molecule, Neurotrophin 4/5, Nerve growth factor, Brain-derived neurotrophic factor, glial cell line-derived neurotrophic factor, ciliary neurotrophic factor and Activity dependent neurotrophic factor.
8. The method of claim 1 , wherein said anode array and said cathode array are separated by between about 10 μm and about 5 mm.
9. The method of claim 8 , wherein said anode array and said cathode array have a length of 0.35 mm to 2.5 mm.
10. The method of claim 1 , wherein said anode array and said cathode array are separated by at least 10 μm.
11. The method of claim 1 , wherein a voltage applied between said anode array and said cathode array is about 1V to about 40V.
12. The method of claim 1 , wherein a total electric charge delivered during said electroporation is less than 0.5 Coulombs, optionally less than 0.1 Coulombs, more optionally less than 0.05 Coulombs.
13. The method of claim 12 , wherein the total electric charge is delivered through less than 100 electric pulses, optionally through 10 or fewer pulses, 5 or fewer pulses, 4 pulses, 3 pulses, 2 pulses or 1 pulse.
14. The method of claim 13 , wherein each pulse is from about 100 μs to about 500 ms in duration.
15. The method of claim 1 , wherein said anode array and said cathode array are provided on a cochlear implant electrode array.
16. The method of claim 2 , wherein said anode array and said cathode array are separated by at least 10 μm.
17. The method of claim 2 , wherein a voltage applied between said anode array and said cathode array is about 1V to about 40V.
18. The method of claim 2 , wherein a total electric charge delivered during said electroporation is less than 0.5 Coulombs, optionally less than 0.1 Coulombs, more optionally less than 0.05 Coulombs.
19. The method of claim 18 , wherein the total electric charge is delivered through less than 100 electric pulses, optionally through 10 or fewer pulses, 5 or fewer pulses, 4 pulses, 3 pulses, 2 pulses or 1 pulse.
20. The method of claim 19 , wherein each pulse is from about 100 μs to about 500 ms in duration.