title: DNA nanotherapy for pre-neoplastic cervical lesions.
abstract: OBJECTIVE
  This study aims to test the hypothesis that targeted nanoparticle delivery of DNA encoding HPV16-regulated diphtheria toxin (DT-A) will result in the death of HPV16-infected cells.
  
  
  METHODS
  Plasmid constructs containing a HPV16 Long Control Region (LCR) DNA sequence upstream of DT-A or luciferase reporter (Luc) DNA sequences were used to formulate poly(β-amino ester) nanoparticles. The effect on tumor growth of HPV/DT-A-nanoparticle injection directly into HPV16(+) CaSki human cervical cancer cell-derived xenografts in mice was determined. To evaluate the ability of the HPV16 LCR regulatory sequence to activate gene expression specifically in HPV16-infected cells, mice underwent bioluminescent optical imaging following intraperitoneal injection of HPV/Luc-nanoparticles. The use of Lutrol F127, a thermal-sensitive gel, to target delivery of nanoparticles and subsequent gene expression to cervical epithelial cells was evaluated in ex vivo cultures of mouse cervix and following intravaginal delivery of nanoparticle/gel in mice, as well as in ex vivo cultures of surgical LEEP samples.
  
  
  RESULTS
  The selected HPV16 LCR regulatory sequence activates gene expression in both HPV16-infected cells and non-infected cells. However, in the cervix, it is specifically active in epithelial cells. Following exposure of cervical cells to HPV/DT-A-nanoparticles mixed with Lutrol F127 gel, DT-A is expressed and cells die.
  
  
  CONCLUSIONS
  An HPV16 DNA sequence that targets gene expression specifically to HPV16-infected cells remains to be discovered. Topical application of a Lutrol F127 thermal gel/nanoparticle mix is illustrative of how to restrict exposure of cells to therapeutic nanoparticles, thereby allowing for targeted DNA delivery to cervical pre-cancerous lesions.
authors.affiliation: Lankenau Institute for Medical Research, 100 Lancaster Ave., Wynnewood, PA 19096, USA. Electronic address: pengw@mlhs.org.
authors.forename: Weidan
authors.initials: W
authors.lastname: Peng
authors.affiliation: Lankenau Institute for Medical Research, 100 Lancaster Ave., Wynnewood, PA 19096, USA; Lankenau Medical Center, 100 Lancaster Ave., Wynnewood, PA 19096, USA. Electronic address: duntonc@mlhs.org.
authors.forename: Charles
authors.initials: C
authors.lastname: Dunton
authors.affiliation: Lankenau Institute for Medical Research, 100 Lancaster Ave., Wynnewood, PA 19096, USA; Lankenau Medical Center, 100 Lancaster Ave., Wynnewood, PA 19096, USA. Electronic address: holtzd@mlhs.org.
authors.forename: David
authors.initials: D
authors.lastname: Holtz
authors.affiliation: Lankenau Medical Center, 100 Lancaster Ave., Wynnewood, PA 19096, USA. Electronic address: mmparva@gmail.com.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Parva
authors.affiliation: Lankenau Medical Center, 100 Lancaster Ave., Wynnewood, PA 19096, USA. Electronic address: stamplerk@mlhs.org.
authors.forename: Kate
authors.initials: K
authors.lastname: Stampler
authors.affiliation: Lankenau Institute for Medical Research, 100 Lancaster Ave., Wynnewood, PA 19096, USA. Electronic address: mff204@gmail.com.
authors.forename: Mark
authors.initials: M
authors.lastname: Forwood
authors.affiliation: Lankenau Institute for Medical Research, 100 Lancaster Ave., Wynnewood, PA 19096, USA; Lankenau Medical Center, 100 Lancaster Ave., Wynnewood, PA 19096, USA. Electronic address: rpgogoi@geisinger.com.
authors.forename: Radhika
authors.initials: R
authors.lastname: Gogoi
authors.affiliation: Veterans Affairs Medical Center, 601 Highway 6 west, Iowa City, IA 52242, USA; Carver College of Medicine, University of Iowa, 451 Newton Rd., 200 Medicine Admst. Bldg., Iowa City, IA 52242, USA. Electronic address: Michael-lace@uiowa.edu.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Lace
authors.affiliation: Massachusetts Institute of Technology, 77 Massachusetts Ave., Bldg. 76, Rm. 653, Cambridge, MA 02142, USA. Electronic address: dgander@mit.edu.
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Anderson
authors.affiliation: Lankenau Institute for Medical Research, 100 Lancaster Ave., Wynnewood, PA 19096, USA; Thomas Jefferson University, Kimmel Cancer Center, 925 Chestnut St., Philadelphia, PA 19107, USA. Electronic address: sawickij@mlhs.org.
authors.forename: Janet A
authors.initials: JA
authors.lastname: Sawicki
mesh_terms: D000818:Animals; D004247:DNA / Q000008:administration & dosage*; D004167:Diphtheria Toxin / Q000235:genetics*; D005260:Female; D052162:Human papillomavirus 16 / Q000235:genetics*; D006801:Humans; D051379:Mice; D053758:Nanoparticles / Q000008:administration & dosage*; D010446:Peptide Fragments / Q000235:genetics*; D011230:Precancerous Conditions / Q000628:therapy*; D002583:Uterine Cervical Neoplasms / Q000628:therapy*
chemical_list: D004167:Diphtheria Toxin; D010446:Peptide Fragments; C029795:diphtheria toxin fragment A; D004247:DNA