Methods of regulating uptake and transcellular transport of leukocytes and therapeutics

Abstract

Methods for controlling and regulating engulfment, uptake and/or transcellular transport at a stage following initial engagement of an agent to the endothelium are provided, based on the identification of CAM-mediated endocytosis and the sphingomyelin/ceramide pathway as active steps in transcellular TEM. Administration of regulators relating to the identified pathways, such as NHE1, sphingomyelinases, acid sphingomyelinase and ceramide, permit control and regulation of uptake and transcellular transport. Control and regulation of uptake and/or transcellular transport is applicable in strategies to modulate inflammation, provide controlled and/or targeted delivery of agents, control pathogenic invasion, recover action of an inhibited CAM-mediated uptake or transendothelial pathway, or provide uptake or transendothelial transport by targeting cell surface markers other than ICAM-1.

What is claimed is: 
     
         1 . A method of potentiating engulfment, uptake or transcellular migration of an agent,
 wherein the agent is complexed to a carrier targeted to a non-ICAM cell surface molecule or receptor,   the method comprising administering a regulator of CAM-mediated endocytosis or the sphingomyelin/ceramide pathway,   said regulator exogenously modulating the non-ICAM cell surface molecule or receptor,   wherein such administration is effective to enhance cellular uptake of the agent and/or transcellular transport of the agent via the non-ICAM cell surface molecule or receptor.   
     
     
         2 . The method of  claim 1 , wherein the non-ICAM cell surface molecule or receptor is associated with a mechanism of endocytosis and/or transport across cells other than CAM-mediated endocytosis. 
     
     
         3 . The method of  claim 2 , wherein the mechanism of endocytosis and/or transport across cells is phagocytosis. 
     
     
         4 . The method of  claim 2 , wherein the mechanism of endocytosis and/or transport across cells is macropinocytosis. 
     
     
         5 . The method of  claim 2 , wherein the mechanism of endocytosis and/or transport across cells is caveolar-mediated transport. 
     
     
         6 . The method of  claim 2 , wherein the mechanism of endocytosis and/or transport across cells is clathrin-mediated transport. 
     
     
         7 . The method of  claim 6 , wherein the carrier is targeted to M6PR and the regulator is not acid sphingomyelinase. 
     
     
         8 . The method of  claim 1 , wherein the agent is selected from the group consisting of autologous white blood cells, foreign white blood cells, pathogens, drugs, research probes, analytical probes, molecular probes, diagnostic agents, therapeutic agents, biologically active agents, research agents, analytical agents, imaging agents, monitoring agents, enzymes, proteins, peptides, hormones, lipids, sugars, nucleic acids, lipoproteins, and chemicals. 
     
     
         9 . The method of  claim 1 , wherein the agent is a biologically active agent. 
     
     
         10 . The method of  claim 9 , wherein the carrier comprises a polymer. 
     
     
         11 . The method of  claim 1 , wherein the regulator is not insulin. 
     
     
         12 . The method of  claim 11 , wherein the regulator is selected from the group consisting of proteins affecting the CAM-mediated pathway, sphingomyelinases, activators of sphingomyelinases, NHE1, lipids affecting the CAM-mediated pathway, sphingomyelin, ceramide, and ceramidases. 
     
     
         13 . The method of  claim 12 , wherein the regulator is selected from the group consisting of NHE1, sphingomyelinases and ceramide. 
     
     
         14 . The method of  claim 1 , wherein the regulator is administered to a patient in need of such administration and the administration of the regulator provides effective levels of the regulator to the endothelium of the patient. 
     
     
         15 . The method of  claim 14 , wherein the administration is parenteral administration, intraperitoneal administration, intravenous administration, intraarterial administration, intradermal administration, intramuscular administration or subcutaneous administration. 
     
     
         16 . A method of potentiating engulfment, uptake or transcellular migration of an agent,
 wherein the agent is complexed to a carrier targeted to a non-ICAM cell surface molecule or receptor, said non-ICAM cell surface molecule or receptor associated with a phagocytosis, macropinocytosis, clathrin-mediated or caveolar-mediated transport pathway,   the method comprising administering neutral sphingomyelinase in vitro to a system or in vivo to a patient in need of such administration,   wherein such administration is effective to enhance the engulfment, uptake and/or transcellular transport of the agent via the non-ICAM cell surface molecule or receptor.   
     
     
         17 . The method of  claim 16 , wherein the administration is effective to enhance cellular uptake of the agent and/or transcellular transport of the agent via the non-ICAM cell surface molecule or receptor. 
     
     
         18 . The method of  claim 16 , wherein said neutral sphingomyelinase exogenously modulates the non-ICAM cell surface molecule or receptor. 
     
     
         19 . The method of  claim 16 , wherein said non-ICAM cell surface molecule or receptor is associated with a phagocytosis transport pathway. 
     
     
         20 . The method of  claim 16 , wherein said non-ICAM cell surface molecule or receptor is associated with a macropinocytosis transport pathway. 
     
     
         21 . The method of  claim 16 , wherein said non-ICAM cell surface molecule or receptor is associated with a clathrin-mediated transport pathway. 
     
     
         22 . The method of  claim 16 , wherein said non-ICAM cell surface molecule or receptor is associated with a caveolar-mediated transport pathway.