Pulsed electric field transfer of molecules to cells while in the body

Abstract

Devices, systems and methods are provided for delivering molecules, particularly small molecules and/or macromolecules, to cells within the body, particularly to target cells which directly therapeutically benefit from the functionality of the molecules. Example molecules include DNA plasmids, RNAs (e.g. messenger RNA (mRNA), small interfering RNA (siRNA), micro RNA), oligonucleotides, antisense oligonucleotides (ASO), proteins and/or materials which invoke genetic or epigenetic changes in the cellular behavior, to name a few. In such instances, the molecules are driven into the target cells with the use of pulsed electric fields (PEFs) which deliver the molecules through the cell wall of the target cells so that the desired genes are able to carry out the desired effect within the cells. Thus, the molecules are to be delivered to the desired location within the body at a desired time and in a desired concentration relative to the delivery of the pulsed electric fields for optimal outcomes. The molecules may be delivered to the body by a variety of methods, such as systemically, regionally and/or by direct injection to the area of the target cells. The pulsed electric field energy is delivered to the target cells in vivo and in situ which drives the molecules into the cells. Thus, the molecules are passed into the cells without the use of viruses or ex vivo methods, such as in vitro electroporation.

1 . (canceled) 
     
     
         2 . A method of transferring molecules to cells of target tissue within a body of a patient comprising:
 delivering a plurality of molecules to the body of the patient;   positioning at least one energy delivery body of an energy delivery device into the body of the patient within sufficient range of the target tissue so as to receive pulsed electric field energy delivered therefrom; and   delivering the pulsed electric field energy to the at least one energy delivery body so as to transmit the pulsed electric field energy to the target tissue in a manner which causes at least one of the plurality of molecules to enter at least one of the cells of the target tissue, wherein the target tissue cells directly therapeutically benefit from the functionality of the molecules.   
     
     
         3 . A method as in  claim 2 , wherein directly therapeutically benefit comprises treatment of a disorder. 
     
     
         4 . A method as in  claim 3 , wherein the disorder comprises a genetic disorder. 
     
     
         5 . A method as in  claim 2 , wherein the at least one molecule of the plurality of molecules comprises a plasmid, DNA, a synthetic DNA vector, RNA, a nucleic acid-based molecule, an antisense oligonucleotide, an oligomer molecule, a ribozyme, a ribonucleoprotein, CRISPR, a recombinant protein, a proteolysis targeting chimera, Zinc Finger Nucleases or Transcription Activator-Like Effector Nucleases, a protein and/or material which invokes genetic or epigenetic changes in cellular behavior. 
     
     
         6 . A method as in  claim 2 , wherein the at least one molecule of the plurality of molecules comprises a gene larger than 10 kb. 
     
     
         7 . A method as in  claim 2 , wherein delivering the plurality of molecules to the body comprises delivering the molecules intravenously to the body. 
     
     
         8 . A method as in  claim 7 , wherein delivering the plurality of molecules to the body comprises delivering the molecules intravenously to the body and locally to the target tissue. 
     
     
         9 . A method as in  claim 2 , wherein delivering the plurality of molecules occurs at least prior to delivering the energy. 
     
     
         10 . A method as in  claim 2 , wherein delivering the plurality of molecules comprises delivering the plurality of molecules to multiple locations within or near the target tissue. 
     
     
         11 . A method as in  claim 10 , wherein the multiple locations are within 0.5 mm-5 cm of the at least one cell of the target tissue. 
     
     
         12 . A method as in  claim 10 , wherein delivering the plurality of molecules at multiple locations comprises delivering different concentrations of molecule solution, different volumes of molecule solution and/or different types of molecule solution to one or more of the multiple locations. 
     
     
         13 . A method as in  claim 10 , wherein the energy delivery device comprises a shaft having one or more tines extendable therefrom, and wherein delivering the plurality of molecules at multiple locations is achieved by delivering molecules through one or more of the one or more tines. 
     
     
         14 . A method as in  claim 13 , wherein the delivering the plurality of molecules comprises delivering different molecules through at least one of the one or more tines in comparison to a least another of the one or more tines. 
     
     
         15 . A method as in  claim 2 , wherein the energy delivery device comprises a shaft having one or more tines extendable therefrom, and wherein the at least one energy delivery body comprises at least one of the one or more tines, and wherein delivering the energy comprises energizing at least one of the plurality of tines. 
     
     
         16 . A method as in  claim 15 , wherein energizing at least one of the plurality of tines comprises individually energizing at least one of the plurality of tines while at least one of the plurality of tines is not energized. 
     
     
         17 . A method as  claim 2 , further comprising inducing extravasation of fluid within a localized area in the body so as to increase delivery of molecules to the target tissue. 
     
     
         18 . A method as in  claim 17 , wherein inducing extravasation occurs prior to delivering energy. 
     
     
         19 . A method as in  claim 17 , wherein delivering the plurality of molecules comprises delivering the plurality of molecules to vasculature of the body and wherein the extravasation is from the vasculature. 
     
     
         20 . A method as in  claim 17 , wherein inducing extravasation comprises delivering conditioning energy to the at least one energy delivery body and wherein the conditioning energy is from another electric signal comprising a plurality of monophasic pulses each having a duration exceeding 500 microseconds. 
     
     
         21 . A method as in  claim 2 , further comprising delivering conditioning energy to the target tissue prior to delivering the pulsed electric field energy, wherein the conditioning energy increases cellular resistance of the target tissue to eventual cell death. 
     
     
         22 . A method as in  claim 2 , further comprising pre-warming the target tissue prior to delivering the pulsed electric field energy. 
     
     
         23 . A method as in  claim 2 , wherein the pulsed electric field energy is from an electric signal comprising a series of pulses, wherein the series of pulses includes at least one pulse having a positive amplitude and at least one pulse of having a negative amplitude. 
     
     
         24 . A method as in  claim 23 , wherein together the series of pulses have a balance of charge from positive amplitude on-time and negative amplitude on-time. 
     
     
         25 . A method as in  claim 24 , wherein together the series of pulses have sufficient balance of charge from positive amplitude on-time and negative amplitude on-time so as to avoid muscle stimulation within the body. 
     
     
         26 . A method as in  claim 24 , wherein together the series of pulses have sufficient balance of charge from positive amplitude on-time and negative amplitude on-time so as to avoid ablation of the target tissue cells.