Abstract A49: Differential expression of the complement regulatory protein CD55 in wildtype and mutant p53 Glioblastoma and Ewing's Sarcoma
Authors: Pandya Pankita H. , Saadatzadeh Reza , Ding Jixin , Bailey Barbara , Ross Sydney , Murray Mary E. , Renbarger Jamie L. , Pollok Karen E. 
Published: March 2017

Background/Objectives: Solid tumors such as Glioblastoma (GBM) and Ewing's Sarcoma result in relapse or refractory disease due the tumor's ability to develop resistance to anti-cancer therapies. One mechanism by which solid tumors such as GBM and Ewing's Sarcoma, circumvent the immuno-editing process and result in resistance to treatments is by the up-regulation of membrane-bound complement regulatory proteins (mCRPs) CD46, CD55, and CD59. While it has been reported that mCRPs are up-regulated in pediatric liquid tumors and influence the efficacy of monoclonal antibody treatments, regulation of mCRP expression in solid tumors has not been explored in detail. To delineate potential mechanisms regulating expression of mCRPs, we first screened wildtype (wt-p53) or mutant p53 solid tumor cell lines for mCRP expression. Our preliminary data suggest that p53 status correlates with CD55 but not CD46 or CD59 expression. These studies may serve to be the foundation for potentially recognizing the mCRPs as immune biomarkers in pediatric and adult solid tumors, ultimately, resulting in the development of novel immunotherapies for improved clinical outcomes and patient care.

Design/Methods: Adult GBM and pediatric Ewing's Sarcoma cell lines that are wildtype p53 (GBM10 and CHLA-9) or mutant p53 (GBM-43 and CHLA-10) were exploited for in vitro studies. The GBM-43 was generated from a primary GBM while the GBM-10 from a patient with recurrent GBM. The paired Ewing's Sarcoma cell lines, CHLA-9 and CHLA-10, were generated from the same patient at primary diagnosis and at relapse respectively. Western blot and qPCR were used to confirm expression and mutational status of p53. mCRP expression was evaluated using RT-PCR and flow cytometry.

Results: All cancer cell lines expressed p53 to varying degrees. No significant difference was observed in the expression of CD46 and CD59 among the wt-p53 and mutant p53 glioblastoma cell lines. Mutant p53 cell lines from glioblastoma and Ewing's sarcoma had increased CD55 transcript levels compared to their wildtype counterpart cell lines where the transcripts were undetectable. In addition, flow cytometry data revealed increased expression of the mCRP, CD55, in mutant p53 glioblastoma (GBM-43) versus wt-p53 (GBM-10) cells (p&lt;0.001); flow cytometric analysis of mCRPs in Ewing's sarcoma cell lines is in progress.

Conclusion: These preliminary findings highlight the importance of further investigating the role of mCRPs on wt-p53 and mutant p53 cell lines. Studies are in progress to determine if expression of CD55 is regulated by p53 status. Understanding how this critical mCRP is regulated in solid tumors will be important for immune biomarker development as well as useful in guiding the use of antibody-based therapeutic approaches in solid tumors.

Citation Format: Pankita H. Pandya, Reza Saadatzadeh, Jixin Ding, Barbara Bailey, Sydney Ross, Mary E. Murray, Jamie L. Renbarger, Karen E. Pollok. Differential expression of the complement regulatory protein CD55 in wildtype and mutant p53 Glioblastoma and Ewing's Sarcoma. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2016 Oct 20-23; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2017;5(3 Suppl):Abstract nr A49.