title: Enhanced immunoPET of ALCAM-positive colorectal carcinoma using site-specific ⁶⁴Cu-DOTA conjugation.
abstract: Activated leukocyte cell adhesion molecule (ALCAM) is an immunoglobulin superfamily cell adhesion molecule that is aberrantly expressed in a wide variety of human tumors, including melanoma, prostate cancer, breast cancer, colorectal carcinoma, bladder cancer and pancreatic adenocarcinoma. This wide spectrum of human malignancies makes ALCAM a prospective pan-cancer immunoPET target to aid in detection and diagnosis in multiple malignancies. In this study, we assess site-specific versus non-site-specific conjugation strategies for (64)Cu-DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid) immunoPET imaging of a fully human ALCAM cys-diabody (cDb) with a reduced linker length that retains its bivalent binding ability. ALCAM constructs with linker lengths of eight, five and three amino acids were produced to make true non-covalent site-specifically modified cDbs. Characterization by gel electrophoresis, size exclusion chromatography, flow cytometry and mass spectrometry of the various constructs was performed. To demonstrate the increased utility of targeting multiple malignancies expressing ALCAM, we compare the targeting of the site-specific versus non-site-specific conjugated cDbs to the human colorectal cancer xenograft LS174T. Interestingly, the conjugation strategy not only affects tumor targeting but also hepatic and renal uptake/clearance.
authors.affiliation: Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA.
authors.forename: Richard
authors.initials: R
authors.lastname: Tavaré
authors.affiliation: Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA.
authors.forename: Wei H
authors.initials: WH
authors.lastname: Wu
authors.affiliation: Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA.
authors.forename: Kirstin A
authors.initials: KA
authors.lastname: Zettlitz
authors.affiliation: Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA.
authors.forename: Felix B
authors.initials: FB
authors.lastname: Salazar
authors.affiliation: Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA.
authors.forename: Katelyn E
authors.initials: KE
authors.lastname: McCabe
authors.affiliation: Department of Anesthesia, UCSF, San Francisco General Hospital, San Francisco, CA 94110, USA.
authors.forename: James D
authors.initials: JD
authors.lastname: Marks
authors.affiliation: Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA awu@mednet.ucla.edu.
authors.forename: Anna M
authors.initials: AM
authors.lastname: Wu
mesh_terms: D020410:Activated-Leukocyte Cell Adhesion Molecule / Q000737:chemistry*; D000818:Animals; D045744:Cell Line, Tumor; D015179:Colorectal Neoplasms / Q000737:chemistry / Q000000981:diagnostic imaging* / Q000378:metabolism; D003300:Copper; D006573:Heterocyclic Compounds, 1-Ring; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D057054:Molecular Imaging / Q000379:methods; D049268:Positron-Emission Tomography / Q000379:methods*; D051381:Rats; D014018:Tissue Distribution
chemical_list: D020410:Activated-Leukocyte Cell Adhesion Molecule; D006573:Heterocyclic Compounds, 1-Ring; C071349:1,4,7,10-tetraazacyclododecane- 1,4,7,10-tetraacetic acid; D003300:Copper
keywords: ALCAM; Cu-64; DOTA; immunoPET; site-specific conjugation
references.citation: Bioconjug Chem. 2008 Dec;19(12):2527-34
references.pmid: 19053310
references.citation: PLoS One. 2013 Aug 01;8(8):e70028
references.pmid: 23936372
references.citation: J Immunother. 2007 May-Jun;30(4):396-405
references.pmid: 17457214
references.citation: Transl Res. 2008 Mar;151(3):122-8
references.pmid: 18279810
references.citation: Methods Enzymol. 1981;73(Pt B):3-46
references.pmid: 7300683
references.citation: J Mol Med (Berl). 2007 Oct;85(10):1113-23
references.pmid: 17554518
references.citation: Cancer Res. 2004 Jan 15;64(2):704-10
references.pmid: 14744788
references.citation: J Nucl Med. 2010 Jul;51(7):1049-58
references.pmid: 20554737
references.citation: J Clin Immunol. 2006 Jan;26(1):12-21
references.pmid: 16418799
references.citation: J Exp Med. 1995 Jun 1;181(6):2213-20
references.pmid: 7760007
references.citation: Protein Eng. 1997 Apr;10(4):423-33
references.pmid: 9194168
references.citation: Protein Eng. 1994 Aug;7(8):1017-26
references.pmid: 7809028
references.citation: Clin Cancer Res. 2008 Mar 15;14(6):1726-33
references.pmid: 18347173
references.citation: J Exp Med. 1995 Apr 1;181(4):1563-8
references.pmid: 7535342
references.citation: Am J Pathol. 2000 Mar;156(3):769-74
references.pmid: 10702391
references.citation: Cancer Metastasis Rev. 2005 Jun;24(2):223-36
references.pmid: 15986133
references.citation: Angew Chem Int Ed Engl. 2009;48(38):6974-98
references.pmid: 19714693
references.citation: Bioconjug Chem. 2001 Mar-Apr;12(2):220-8
references.pmid: 11312683
references.citation: Protein Eng Des Sel. 2004 Jan;17(1):21-7
references.pmid: 14985534
references.citation: Curr Med Chem. 2010;17(24):2636-55
references.pmid: 20491631
references.citation: Bioconjug Chem. 2009 Nov;20(11):2071-81
references.pmid: 19874007
references.citation: Blood. 2014 Jul 24;124(4):519-29
references.pmid: 24740813
references.citation: J Med Chem. 2013 Jun 27;56(12):4966-74
references.pmid: 23692562
references.citation: Mol Imaging Biol. 2012 Jun;14(3):336-47
references.pmid: 21630083
references.citation: FEBS Lett. 1999 Jun 18;453(1-2):164-8
references.pmid: 10403395
references.citation: Blood. 2010 Jan 21;115(3):736-44
references.pmid: 19903902
references.citation: Biopolymers. 2008;90(3):450-8
references.pmid: 17618518
references.citation: Cancer Immunol Immunother. 2010 Nov;59(11):1665-74
references.pmid: 20635083
references.citation: Breast Cancer Res. 2004;6(5):R478-87
references.pmid: 15318930
references.citation: Nat Biotechnol. 2005 Sep;23(9):1137-46
references.pmid: 16151407
references.citation: J Clin Pathol. 2004 Nov;57(11):1160-4
references.pmid: 15509676
references.citation: Cancer Biother Radiopharm. 2009 Aug;24(4):379-93
references.pmid: 19694573
references.citation: J Nucl Med. 2009 Mar;50(3):435-43
references.pmid: 19223400
references.citation: Bioconjug Chem. 2002 Sep-Oct;13(5):985-95
references.pmid: 12236780
references.citation: J Clin Oncol. 2012 Nov 1;30(31):3884-92
references.pmid: 22987087
references.citation: J Proteomics. 2014 Mar 17;99:138-51
references.pmid: 24487095
references.citation: Exp Eye Res. 2012 Nov;104:89-93
references.pmid: 22940369
references.citation: Cancer Biother Radiopharm. 2008 Aug;23(4):411-23
references.pmid: 18771345
references.citation: Protein Eng Des Sel. 2010 Apr;23(4):243-9
references.pmid: 20053640
references.citation: Bioconjug Chem. 2009 Aug 19;20(8):1474-81
references.pmid: 19642689
references.citation: Mol Imaging. 2003 Jul;2(3):131-7
references.pmid: 14649056
references.citation: J Nucl Med. 2009 Jan;50(1):2-5
references.pmid: 19091888
references.citation: Q J Nucl Med. 1998 Dec;42(4):262-70
references.pmid: 9973841
references.citation: J Pathol. 2005 Feb;205(3):359-76
references.pmid: 15532095
references.citation: Methods. 2014 Jan 1;65(1):139-47
references.pmid: 24091005
references.citation: Nat Biotechnol. 2008 Aug;26(8):925-32
references.pmid: 18641636
references.citation: Annu Rev Biochem. 2010;79:413-44
references.pmid: 20307192
references.citation: J Nucl Med. 2006 Sep;47(9):1546-53
references.pmid: 16954565
references.citation: Gastroenterology. 2005 Oct;129(4):1187-97
references.pmid: 16230073