Doxorubicin-loaded Fab' fragments of anti-disialoganglioside immunoliposomes selectively inhibit the growth and dissemination of human neuroblastoma in nude mice.
Neuroblastoma (NB) is the most common extracranial solid tumor in children. Intensive therapeutic intervention does not prolong the overall disease-free survival rate for this tumor. NB tumor, but not normal tissues, overexpress the disialoganglioside (GD(2)) at the cell surface. Anti-GD(2) whole antibodies (aGD(2)) or their corresponding Fab' fragments were covalently coupled to Stealth immunoliposomes (aGD(2)-SIL or Fab'-SIL), and their binding to GD(2)-positive NB cells was measured. Cytotoxic effects of immunoliposomes loaded with doxorubicin (DXR) were determined. Radiolabelled immunoliposomes were used to evaluate pharmacokinetics (PK). The effectiveness of different liposomal formulations of DXR was tested against a metastatic model of human NB in nude mice. aGD(2)-SIL and Fab'-SIL showed concentration-dependent specific binding and uptake by GD(2)-positive NB cells. DXR entrapped in aGD(2)-SIL or Fab'-SIL (aGD(2)-SIL[DXR], Fab'-SIL[DXR]) showed higher cytotoxicities than nontargeted liposomes (SL[DXR]). DXR-loaded Fab'-SIL (Fab'-SIL[DXR]) also showed specific binding, uptake, and cytotoxic effects on several GD(2)-positive NB cells in vitro. PK studies showed that Fab'-SIL had long-circulating profiles in blood compared with aGD(2)-SIL, with the PK profile for Fab'-SIL being almost identical to that obtained with nontargeted Stealth liposomes. In vivo, long-term survivors were obtained in mice treated with Fab'-SIL[DXR] but not in untreated animals, or those treated with free aGD(2) Fab', Fab'-SIL (no drug), free-DXR, or nontargeted Stealth liposomes[DXR] (no antibody; P < 0.0001). Immunoliposomes containing DXR prevented the establishment and growth of the tumor in all of the organs examined. In conclusion, Fab'-SIL[DXR] formulations led to the total inhibition of metastatic growth of human NB in a nude mouse metastatic model. This formulation should receive clinical evaluation as adjuvant therapy of NB.
Pastorino
Fabio
F
Differentiation Therapy Unit, Laboratory of Oncology, G. Gaslini Children's Hospital, Genoa, Italy 16148.
Brignole
Chiara
C
Marimpietri
Danilo
D
Sapra
Puja
P
Moase
Elaine H
EH
Allen
Theresa M
TM
Ponzoni
Mirco
M
D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects*; D004317:Doxorubicin / Q000008:administration & dosage* / Q000627:therapeutic use; D004337:Drug Carriers; D005732:Gangliosides / Q000276:immunology*; D006801:Humans; D007140:Immunoglobulin Fab Fragments; D008081:Liposomes; D051379:Mice; D008819:Mice, Nude; D009447:Neuroblastoma / Q000188:drug therapy* / Q000473:pathology*; D014407:Tumor Cells, Cultured
D004337:Drug Carriers; D005732:Gangliosides; D007140:Immunoglobulin Fab Fragments; D008081:Liposomes; C025447:sialogangliosides; D004317:Doxorubicin