METHOD AND DEVICES FOR IMPROVED EFFICIENCY OF RNA DELIVERY TO CELLS
First Claim
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1. A method of increasing an uptake of an ribonucleic acid sequence by a cell, said cell being selected from the group consisting of cardiac cells, skeletal muscle cells, kidney cells, neurons and glial cells, the method comprising:
- a) applying a plurality of pulses of an electric field to the cell for a time period between about two and about 24 hours;
b) within 24 hours of the step a), administering the ribonucleic acid sequence to the cellwherein;
the electric field has strength of between about 0.5 V/cm and about 40 V/cm, calculated according to Formula I;
E=V/d wherein in said formula E is the strength, V is Voltage and d is distance between electrodes.
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Abstract
The instant invention provides a method for improving efficiency of RNA delivery to cells. The method comprises applying a low strength electric field to the cells and then after a certain time period, administering the ribonucleic acid sequence to the cells. Devices, kits, and RNA molecules suitable for delivery and devices suitable for practicing the disclosed methods are also provided.
24 Citations
49 Claims
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1. A method of increasing an uptake of an ribonucleic acid sequence by a cell, said cell being selected from the group consisting of cardiac cells, skeletal muscle cells, kidney cells, neurons and glial cells, the method comprising:
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a) applying a plurality of pulses of an electric field to the cell for a time period between about two and about 24 hours; b) within 24 hours of the step a), administering the ribonucleic acid sequence to the cell wherein; the electric field has strength of between about 0.5 V/cm and about 40 V/cm, calculated according to Formula I;
E=V/dwherein in said formula E is the strength, V is Voltage and d is distance between electrodes. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27)
placing a plurality of electrodes into said pre-determined area prior to applying the plurality of pulses.
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14. The method of claim 13, wherein the placement of the electrodes results in a pre-determined shape of the electric field.
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15. The method of claim 10, wherein the nucleic acid is administered to the pre-determined area through a catheter.
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16. The method of claim 15, further comprising a step of verifying the placement of the catheter.
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17. The method of claim 10, further comprising a step of verification of the placement of the electrodes.
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18. The method of claim 1, wherein the cell is a heart cell, and wherein the strength is between about 2 V/cm and about 12 V/cm, and wherein said the members of the plurality of pulses are administered as bursts having frequency of about ten pulses, said bursts separated by about 500 ms, and wherein the time period is about 20 hours.
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19. The method of claim 1, further comprising the step of determining a heart rate of a patient, and wherein
i) the cell is a heart cell, ii) the strength is between about 2 V/cm and about 12 V/cm, iii) the time period is about 20 hours, and wherein said the members of the plurality of pulses, and iv) the frequency of said bursts is about identical to the heart rate of the patient. -
20. The method of claim 19, wherein an onset of a heart beat initiates the application of the burst of pulses of the electric field.
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21. The method of claim 18, wherein the siRNA or the shRNA inhibits expression of phospholamban.
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22. The method of claim 1, wherein the cell is a kidney cell, and wherein the strength is between about 0.5 and about 40 V/cm and the time period is between about one and about 12 hours.
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23. The method of claim 22, wherein the time period is about two hours.
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24. The method of claim 22, wherein the siRNA or the shRNA inhibits expression of BIM.
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25. The method of claim 1, wherein the ribonucleic acid sequence is administered in a composition comprising an imaging agent, the method further comprising:
- determining a distribution of the composition.
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26. The method of claim 18, wherein
i) said plurality of pulses of an electric field are applied to a pre-determined area within a heart or a kidney of a patient through a plurality of electrodes, ii) said plurality of electrodes are positioned relative to each other as to provide an electric field of a pre-determined shape. -
27. The method of claim 1, wherein step (b) is initiated at least 1 minute, at least 5 minutes, at least 30 minutes, at least 45 minutes, at least 1 hour, at least 2 hours, at least 3 hours, at least 4 hours, at least 5 hours, at least 6 hours, at least 7 hours, at least 8 hours, at least 9 hours, at least 10 hours, at least 11 hours, at least 12 hours, at least 13 hours, at least 14 hours, at least 15 hours, at least 16 hours, at least 17 hours, at least 18 hours, at least 19 hours, at least hours, at least 21 hours, at least 22 hours, at least 23 hours or 24 hours after the initiation of step (a).
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28. A device for delivering a nucleic acid sequence to a cell within a patient, the device comprising:
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a plurality of electrodes; a catheter, the catheter comprising a wall and a cavity; a reservoir containing a composition comprising the RNA, said reservoir fluidly connected with said catheter, wherein said composition is RNAse-free; a pump operably connected to said reservoir; a processor operably connected to the members of said plurality of electrodes and adapted to receive electrical signals from said members and to deliver an electric field to said members. - View Dependent Claims (29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39)
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40. A medical system comprising:
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a plurality of electrodes; a catheter, the catheter comprising a wall and a cavity, wherein the members of said plurality of electrodes are disposed within or on the surface of said wall; a reservoir containing a composition comprising the RNA, said reservoir fluidly connected with said catheter; a pump operably connected to said reservoir; a processor operably connected to the members of said plurality of electrodes and adapted to receive electrical signals from said members and to deliver an electric field to said members; and a patient-specific intraoperative image-guided mapping means.
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41. A kit comprising:
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a) a plurality of electrodes; b) a composition comprising RNA; c) a processor adapted to; i) actuate an electric stimulation by the members of said plurality of electrodes; ii) receive a signal from the members of said plurality of electrodes and, iii) within a predetermined time period after receiving said signal from the members of said plurality of electrodes, to actuate release of at least a portion of said composition comprising RNA. - View Dependent Claims (42, 43, 44, 45, 46, 47, 48, 49)
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Specification