Receptor for the globular heads of C1q (gC1q-R, p33, hyaluronan-binding protein) is preferentially expressed by adenocarcinoma cells.
Combinatorial Ig libraries with phage display allow in vitro generation of human Ig fragments without the need to maintain hybridomas in ongoing cell culture or to select circulating Ig from human serum. Identifying tumor-associated antigens on the surface of intact tumor cells, as opposed to purified proteins, presents a challenge due to the difficulty of preserving complex 3-D epitopic sites on the cell surface, the variable expression of antigens on different malignant cell types and the stereotactic interference of closely associated proteins on the intact membrane surface limiting accessibility to antigenic sites. A combinatorial Ig library of 10(10) clones was generated from the cDNA of PBMCs derived from patients with breast adenocarcinoma. Following subtractive panning, the library was enriched for Ig (Fab fragment) binding to intact adenocarcinoma cells and the resultant Fabs were screened against a cDNA expression library, itself generated from breast cancer cells. Using this approach, we isolated clones from the cDNA library expressing gC1q-R, a glycoprotein comprising the major structure of C1, the first component of the complement system. gC1q-R is a 33 kDa glycoprotein expressed not only on the cell surface but also intracellularly, with motifs that target it to mitochondria and complete homology with HABP and human HeLa cell protein p32, which is copurified with pre-mRNA SF2. Sequencing of the gene encoding tumor-associated gC1q-R did not reveal any consistent tumor-specific mutations. However, histochemical staining with anti-gC1q-R MAb demonstrated marked differential expression of gC1q-R in thyroid, colon, pancreatic, gastric, esophageal and lung adenocarcinomas compared to their nonmalignant histologic counterparts. In contrast, differential expression was not seen in endometrial, renal and prostate carcinomas. Despite high expression in breast carcinoma, gC1q-R was also expressed in nonmalignant breast tissue. Although the precise relation of gC1q-R to carcinogenesis remains unclear, our finding of tumor overexpression and the known multivalent binding of gC1q-R to not only C1q itself but also a variety of circulating plasma proteins as well as its involvement in cell-to-cell interactions suggest that gC1q-R may have a role in tumor metastases and potentially serve in molecule-specific targeting of malignant cells.
Rubinstein
Daniel B
DB
Section of Hematology/Oncology, Boston University School of Medicine, Boston, MA 02118, USA. brurion@bu.edu
Stortchevoi
Alexei
A
Boosalis
Michael
M
Ashfaq
Raheela
R
Ghebrehiwet
Berhane
B
Peerschke
Ellinor I B
EI
Calvo
Fabien
F
Guillaume
Thierry
T
D000230:Adenocarcinoma / Q000378:metabolism*; D001483:Base Sequence; D015152:Blotting, Northern; D001943:Breast Neoplasms / Q000378:metabolism; D002352:Carrier Proteins; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D015922:Complement C1q / Q000096:biosynthesis*; D018076:DNA, Complementary / Q000378:metabolism; D000939:Epitopes; D015723:Gene Library; D006801:Humans; D018960:Hyaluronan Receptors / Q000096:biosynthesis*; D007150:Immunohistochemistry; D008562:Membrane Glycoproteins; D024101:Mitochondrial Proteins; D008954:Models, Biological; D008969:Molecular Sequence Data; D009362:Neoplasm Metastasis; D019151:Peptide Library; D011485:Protein Binding; D012333:RNA, Messenger / Q000378:metabolism; D011951:Receptors, Complement / Q000096:biosynthesis*; D012689:Sequence Homology, Nucleic Acid
C087299:C1QBP protein, human; D002352:Carrier Proteins; D018076:DNA, Complementary; D000939:Epitopes; D018960:Hyaluronan Receptors; D008562:Membrane Glycoproteins; D024101:Mitochondrial Proteins; D019151:Peptide Library; D012333:RNA, Messenger; D011951:Receptors, Complement; C032402:complement 1q receptor; D015922:Complement C1q