SEREX was used to identify candidate tumor antigens in the nonimmunogenic fibrosarcoma (NFSA) tumor model. One of the six clones identified was of particular interest. NFSA-5 was identified as the receptor for hylaronan-acid-mediated motility (RHAMM), which is involved in cell growth and metastasis. RHAMM is expressed in a variety of human tumors. RHAMM is differentially expressed, with significant levels not found in normal tissues other than testis, placenta, and thymus. Therefore, RHAMM may be an appealing target for human tumor vaccines. The identification of murine homologs to human tumor antigens may aid in the preclinical development of human tumor vaccines. The goal of our studies was to use serological analysis of antigens by recombinant expression cloning (SEREX) to identify candidate tumor antigens in a nonimmunogenic murine fibrosarcoma model. SEREX provides a rapid means of identifying candidate tumor antigens in murine cancer models. The identification of murine homologs to human tumor antigens may aid in the preclinical development of human tumor vaccines. The SEREX approach included construction of a cDNA expression library from NFSA tumors followed by immunoscreening of the library with sera from C3H mice growing NFSA tumors. The nucleotide sequence of insert cDNA was determined for positive clones. Sequence alignments were performed with BLAST software on GenBank database. Six positive clones were identified. Two clones coded for proteins with known expression in normal tissues. Two clones represented heat-shock proteins, known to be upregulated in human and murine tumors. Two of the clones were of particular interest. Clone NFSA-1 was the homolog to NY-REN-58, an antigen previously identified by SEREX analysis of renal cell carcinoma patients. NFSA-5 was identified as the receptor for the hylaronan-acid-mediated motility (RHAMM), which is involved in cell growth and metastasis. RHAMM was recently identified as a leukemia-associated antigen and is expressed in a variety of human solid tumors including renal cell carcinoma, breast carcinoma, and ovarian carcinoma. RHAMM is differentially expressed, with significant levels not found in normal tissues other than testis, placenta, and thymus. Therefore, RHAMM may be an appealing target for human tumor vaccines.
To determine whether sacral chordoma is monoclonal or polyclonal in origin, a new assay to study the polymorphic human androgen receptor locus (HUMARA) was applied. The ratio of maternal inactive X-chromosone to the paternal inactive X (Lyon hypothesis) was determined via a methylation-specific polymerase chain reaction (PCR) technique. Seven of seven informative samples showed a polyclonal proliferation pattern. This study suggests that chordomas are more comparable to mesenchymal neoplasms than to monoclonal hematopoeitic neoplasms. The purpose of this study was to determine whether sacral chordoma is monoclonal or polyclonal in origin via a new molecular genetic assay of the x-chromosome. A polyclonal proliferation pattern was identified in all informative samples studied. Characterization of the genetic tumorigenesis of this unpredictable neoplasm may lend insight into its biological behavior and offer novel therapeutic intervention. Utilizing a new assay to study the polymorphic human androgen receptor locus (HUMARA), the ratio of maternal inactive X-chromosome to the paternal inactive X (Lyon hypothesis) is determined via a methylation-specific polymerase chain reaction (PCR) technique to detect X-chromosome polymorphisms. Eight female chordoma patients had their DNA harvested and their x-chromosome inactivation pattern and polymorphisms determined and compared to control. A polyclonal proliferation pattern was identified in seven of seven informative samples. The eighth sample showed a single x chromosome allele in normal and tumor tissue and was thus viewed as uninformative.