Methods for dna-dependent targeting of a cell permeant antibody

Abstract

The invention provides methods for selective targeting of live cells, which have undergone or are undergoing radiation or chemotherapy, at a site of interest with a cell-penetrating polypeptide. In one embodiment of the invention, the method comprises contacting the live cells with a cell-penetrating polypeptide comprising cell-penetrating determinants so that the cell-penetrating polypeptide binds extracellular DNA near or around the live cells so as to form a complex or association therewith such that the complex or associated polypeptide-DNA so bound bind the live cells and penetrates the live cells thereby selectively targeting live cells at a site of interest with a cell-penetrating polypeptide.

1 .- 96 . (canceled) 
     
     
         97 . A method comprising:
 administering a complex near or around live cells at a site of interest, wherein the complex comprises a cell-penetrating polypeptide bound to DNA;   wherein the complex penetrates the live cells at the site of interest,   wherein the cell penetrating polypeptide is a 3E10 antibody or a fragment thereof, and   wherein the fragment thereof is a single chain Fv (scFv) fragment of mAb 3E10 antibody.   
     
     
         98 . The method of  claim 97 , wherein the complex targets the live cells at the site of interest with the cell-penetrating peptide. 
     
     
         99 . The method of  claim 97 , wherein the cell-penetrating polypeptide is conjugated to a therapeutic agent. 
     
     
         100 . The method of  claim 97 , wherein the mAb 3E10 antibody, the fragment of the mAb 3E10 antibody, or the scFv fragment of the mAb 3E10 antibody is humanized. 
     
     
         101 . The method of  claim 97 , wherein the 3E10 antibody, the fragment of the mAb 3E10 antibody, or the scFv fragment of the mAb 3E10 antibody is joined to an amino acid sequence AGIH at its amino terminus. 
     
     
         102 . The method of  claim 97 , wherein the 3E10 antibody, the fragment of the mAb 3E10 antibody, or the scFv fragment of the mAb 3E10 antibody is joined to or comprises a peptide linker which in turn comprises a portion of an immunoglobulin heavy chain constant domain CH1 and a swivel sequence, and optionally, the 3E10 antibody, the fragment of the mAb 3E10 antibody, or the scFv fragment of the mAb 3E10 antibody comprises an amino acid sequence AGIH at its amino terminus. 
     
     
         103 . The method of  claim 97 , wherein the fragment of the mAb 3E10 antibody or the scFv fragment of the mAb 3E10 antibody has a change in its amino acid sequence, wherein the change in the amino acid sequence enhances cell penetration and comprises an amino acid substitution in the 3E10 variable heavy chain at amino acid 31 from an aspartic acid to an asparagine (D31N). 
     
     
         104 . The method of  claim 97 , wherein the 3E10 antibody, the fragment of the mAb 3E10 antibody, or the scFv fragment of the mAb 3E10 antibody is naked. 
     
     
         105 . The method of  claim 97 , wherein cell penetration is dependent on a nucleoside salvage pathway. 
     
     
         106 . The method of  claim 105 , wherein the nucleoside salvage pathway is a pathway mediated by equilibrative nucleoside transporters (ENTs) or SLC29 family of integral membrane proteins. 
     
     
         107 . The method of  claim 106 , wherein the equilibrative nucleoside transporter (ENT) or a member of the SLC29 family of integral membrane proteins is a transporter for purine and pyrimidine nucleosides and nucleobases or a metabolite thereof. 
     
     
         108 . The method of  claim 107 , wherein the transporter for purine and pyrimidine nucleosides and nucleobases or a metabolite thereof is an equilibrative nucleoside transporter ENT2. 
     
     
         109 . The method of  claim 97 , wherein the DNA is single-stranded, double-stranded, triple-stranded, or four-stranded or a combination thereof. 
     
     
         110 . The method of  claim 97 , wherein the DNA is artificial DNA. 
     
     
         111 . The method of  claim 97 , wherein the site of interest is an injury site. 
     
     
         112 . The method of  claim 111 , wherein an injury in the injury site is an intracranial injury, brain injury, myocardial infarction, skin injury, liver injury, gastrointestinal injury, lung injury, eye injury, kidney injury, pancreas injury, peritoneal injury, bone injury, nasopharyngeal injury, uterine injury, cervical injury, breast injury, organ injury, tissue injury, burn or radiation injury. 
     
     
         113 . The method of  claim 112 , wherein an injury in the injury site is an acute renal failure, acute organ failure, liver injury, bowel infarction, peripheral vascular disease, or pulmonary failure. 
     
     
         114 . The method of  claim 113  wherein the brain injury is a brain trauma, spinal cord injury, peripheral nerve injury, or stroke. 
     
     
         115 . The method of  claim 97 , wherein the cell-penetrating polypeptide is further linked or bound to an imaging agent or detectable marker. 
     
     
         116 . The method of  claim 97 , wherein administering the complex near or around the live cells at the site of interest is or comprises administration of the complex at the site of interest by injection, microinjection, microprojectile or implantation or a combination thereof.