title: Fe-bLf nanoformulation targets survivin to kill colon cancer stem cells and maintains absorption of iron, calcium and zinc.
abstract: OBJECTIVE
  To validate the anticancer efficacy of alginate-enclosed, chitosan-conjugated, calcium phosphate, iron-saturated bovine lactoferrin (Fe-bLf) nanocarriers/nanocapsules (NCs) with improved sustained release and ability to induce apoptosis by downregulating survivin, as well as cancer stem cells.
  
  
  METHODS
  The stability, nanotoxicity of the modified nanoformulation was evaluated and their anticancer efficacy was re-examined. Their mechanism of internalization was studied and we identified the role of various miRNAs in absorption of these NCs/iron in various body parts of mice. We determined the effect of these NCs on survivin, stem cell markers, red blood cell count, iron, calcium and zinc concentration in mice, determined the antiangiogenic properties of these NCs and studied their effect on cancer stem-like cells.
  
  
  RESULTS
  Spherical NCs (396.1 ± 27.2 nm) exceedingly reduced viability of Caco-2 cells (32 ± 2.83%). The NCs also showed effective internalization and reduction of cancer stem cell markers in triple-positive CD133, survivin and CD44 cancer stem-like cells. Mice treated with the NCs showed no nanotoxicity and did not develop any tumors in xenograft colon cancer models. We found that the serum iron, zinc and calcium absorption were increased. DMT1, LRP, transferrin and lactoferrin receptors were responsible for internalization of the NCs. Different miRNAs were responsible for iron regulation in different organs. Interestingly, NCs inhibited survivin and its different isoforms.
  
  
  CONCLUSIONS
  Our results confirmed that NCs internalized and changed the expression of selected miRNAs that further enhanced their uptake. The NCs activated both extrinsic, as well as intrinsic apoptotic pathways to induce apoptosis by targeting survivin in cancer cells and cancer stem cells, without inducing any nonspecific nanotoxicity. Apart from inhibiting angiogenesis and stem cell markers, NCs also maintained iron and calcium levels.
authors.affiliation: Nanomedicine, Laboratory of Immunology & Molecular Biomedical Research (LIMBR), School of Medicine (SoM), Faculty of Health, Deakin University, Waurn Ponds, Victoria 3217, Australia.
authors.forename: Jagat R
authors.initials: JR
authors.lastname: Kanwar
authors.forename: Ganesh
authors.initials: G
authors.lastname: Mahidhara
authors.forename: Kislay
authors.initials: K
authors.lastname: Roy
authors.forename: Sreenivasan
authors.initials: S
authors.lastname: Sasidharan
authors.forename: Subramanian
authors.initials: S
authors.lastname: Krishnakumar
authors.forename: Neerati
authors.initials: N
authors.lastname: Prasad
authors.forename: Rakesh
authors.initials: R
authors.lastname: Sehgal
authors.forename: Rupinder K
authors.initials: RK
authors.lastname: Kanwar
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D018938:Caco-2 Cells; D002118:Calcium / Q000378:metabolism; D002417:Cattle; D048271:Chitosan / Q000008:administration & dosage / Q000737:chemistry; D003110:Colonic Neoplasms / Q000188:drug therapy* / Q000473:pathology; D016503:Drug Delivery Systems; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000096:biosynthesis*; D007501:Iron / Q000378:metabolism; D007781:Lactoferrin / Q000008:administration & dosage* / Q000737:chemistry; D051379:Mice; D053769:Nanocapsules / Q000008:administration & dosage* / Q000737:chemistry; D014411:Neoplastic Stem Cells / Q000187:drug effects* / Q000473:pathology; D000077022:Survivin; D015032:Zinc / Q000378:metabolism
chemical_list: C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D053769:Nanocapsules; D000077022:Survivin; D048271:Chitosan; D007501:Iron; D007781:Lactoferrin; D015032:Zinc; D002118:Calcium
keywords: Fe-bLf; absorption; iron metabolism; miRNA; nanocarriers/nanocapsules; nanotoxicity; oral delivery; stem cell; survivin; xenograft