Use of cytoskeleton-specific immunoliposomes for preservation of cell viability and gene delivery

Pathological conditions such as hypoxia and inflammation provoke cell membrane lesions representing sub-microscopic holes through which components of cytoskeleton become exposed to the extracellular mileur. Cytoskeleton-specific antibodies coupled to liposomes can deliver the phospholipid vesicles to such lesions. We have used the in vitro delivery of antimyosin antibody-mediated liposomes to hypoxic cardiomyocytes in order: (1) to prevent cell death by sealing membrane lesions, and (2) to provide a novel method of intracellular delivery of DNA. A hypoxic model of injury in H9C2 rat embryonic cardiocytes was used in our experiments. Treatment with antimyosin-immunoliposomes improved the survival of these hypoxic cells significantly relative to controls. Under this artificially induced stress, cytoskeleton-specific immunoliposomes can also deliver their intraliposomal contents into the cytoplasm of the target cells, providing an excellent opportunity for the development of a method for the targeted intracellular delivery of drugs and genetic constructs. This potential was confirmed by demonstration of efficient intracellular delivery of antimyosin sFc vector, pGL2 luciferase vector and bacterial beta galactosidase vector into hypoxic cells with concomitant restoration of cell viability.