Methods of monitoring the efficacy of anti-CD40 antibodies in treating a subject for a CD40-expressing cancer

A method for identifying a subject having a cancer that is responsive to treatment with an anti-CD40 therapeutic agent, said method comprising:

• a) providing a test biological sample obtained from said subject and a control biological sample obtained from said subject, wherein said test biological sample and said control biological sample comprise CD40-expressing neoplastic cells that have been stimulated with a CD40 ligand;
• b) contacting said test biological sample with an effective amount of said anti-CD40 therapeutic agent;
• c) detecting the level of at least one biomarker in said test biological sample, wherein said biomarker is selected from the group consisting of:
• (i) a biomarker of cellular apoptosis, wherein said biomarker of apoptosis is selected from the group consisting of a cleaved Caspase protein, cleaved poly ADP-ribose polymerase (PARP), cell surface expression of phosphotidylserine (PS), genomic DNA fragmentation, and any combinations thereof;
• (ii) a biomarker of a CD40L-mediated CD40 signaling pathway, wherein said biomarker of said CD40L-mediated CD40 signaling pathway is selected from the group consisting of phospho-PI3K, phospho-PDK1, phospho-AKT, phospho-MEK, phospho-ERK, phospho-p38, phospho-IKKα/β, phospho-IκB protein, and activated NF-κB;
• (iii) and a biomarker of cell survival, wherein said biomarker of cell survival is selected from the group consisting of an anti-apoptotic protein that is a Bcl-2 family member, an IAP apoptosis inhibitor protein, and TNF receptor-associated factor-1 (TRAF-1);
• and
• d) comparing the level of said at least one biomarker in said test biological sample to the level of said at least one biomarker detected in said control biological sample, wherein said control biological sample has not been contacted with said anti-CD40 therapeutic agent, wherein
• (i) at least a 20% increase in the level of at least one of said biomarkers of apoptosis, and/or
• (ii) at least a 25% reduction in the level of at least one of said biomarkers of at least one of said CD40L-mediated CD40 signaling pathways, and/or
• (iii) at least a 25% reduction in the level of at least one of said biomarkers of cell survival
• in said test biological sample relative to said control biological sample is indicative of a subject who would benefit from treatment with said anti-CD40 therapeutic agent;
• wherein said anti-CD40 therapeutic agent is an antagonist anti-CD40 monoclonal antibody that is capable of specifically binding to a human CD40 antigen expressed on the surface of a human B cell, said monoclonal antibody being free of significant agonist activity when bound to the CD40 antigen expressed on the surface of said B cell, wherein said anti-CD40 monoclonal antibody is selected from the group consisting of:
• a) a monoclonal antibody comprising a light chain variable domain containing the complementarity determining region (CDR) residues of SEQ ID NO:2 and a heavy chain variable domain containing the CDR residues of SEQ ID NO:4;
• b) a monoclonal antibody comprising a light chain variable domain containing the CDR residues of SEQ ID NO:2 and a heavy chain variable domain containing the CDR residues of SEQ ID NO:5;
• c) a monoclonal antibody comprising a light chain variable domain containing the CDR residues of SEQ ID NO:6 and a heavy chain variable domain containing the CDR residues of SEQ ID NO:7;
• d) a monoclonal antibody comprising a light chain variable domain containing the CDR residues of SEQ ID NO:6 and a heavy chain variable domain containing the CDR residues of SEQ ID NO:8;
• and
• e) a monoclonal antibody that is an antigen-binding fragment of a monoclonal antibody of any one of preceding items a)-j), wherein said fragment retains the capability of specifically binding to said human CD40 antigen.