Identification of survivin as a promising target for the immunotherapy of adult B-cell acute lymphoblastic leukemia

Abstract

B-cell acute lymphoblastic leukemia (B-ALL) is a rare heterogeneous disease characterized by a block in lymphoid differentiation and a rapid clonal expansion of immature, non-functioning B cells. Adult B-ALL patients have a poor prognosis with less than 50% chance of survival after five years and a high relapse rate after allogeneic haematopoietic stem cell transplantation. Novel treatment approaches are required to improve the outcome for patients and the identification of B-ALL specific antigens are essential for the development of targeted immunotherapeutic treatments.We examined twelve potential target antigens for the immunotherapy of adult B-ALL. RT-PCR indicated that only survivin and WT1 were expressed in B-ALL patient samples (7/11 and 6/11, respectively) but not normal donor control samples (0/8). Real-time quantitative (RQ)-PCR showed that survivin was the only antigen whose transcript exhibited significantly higher expression in the B-ALL samples (n = 10) compared with healthy controls (n = 4)(p = 0.015). Immunolabelling detected SSX2, SSX2IP, survivin and WT1 protein expression in all ten B-ALL samples examined, but survivin was not detectable in healthy volunteer samples. To determine whether these findings were supported by the analyses of a larger cohort of patient samples, we performed metadata analysis on an already published microarray dataset. We found that only survivin was significantly over-expressed in B-ALL patients (n = 215) compared to healthy B-cell controls (n = 12)(p = 0.013).We have shown that survivin is frequently transcribed and translated in adult B-ALL, but not healthy donor samples, suggesting this may be a promising target patient group for survivin-mediated immunotherapy.

Serum profiling identifies ibrutinib as a treatment option for young adults with B‐cell acute lymphoblastic leukaemia

“…Of the other B‐ALL patient samples analysed, two had detectable BMX expression (ALL008 and ALL020 PB), three patients expressed DCTPP1 (ALL007, ALL008 and ALL020 BM) and three patients expressed VGLL4 and survivin (ALL007, ALL008 and ALL020 PB and BM). ALL007 and ALL008 had also been examined by our group previously (Boullosa et al , ) and while ALL007 was found to have detectable survivin transcripts in both studies, ALL008 was negative for detectable survivin transcripts in our previous study. None of the HVs had consistently detectable levels of BMX, while all expressed DCTPP1 and VGLL4.…”

“…We identified three antigens that had been shown to play a significant role in cancer(s) and examined their expression in patient and HV samples. We have previously shown that survivin had significantly higher expression in B‐ALL samples compared with healthy controls by qPCR analysis ( P = 0·015) and gene expression analysis ( P = 0·013) (Boullosa et al , ). However there were no statistically significant differences between the expression of the four antigens examined in this study (BMX, DCTPP1, survivin or VGLL4) in patient samples compared with HVs by qPCR.…”

“…We determined the relative expression of transcripts encoding the antigens of interest in the following cell lines that we used as controls (K562, HEK293T, MDA‐MB‐231 and MD‐MB‐468) (Table ). We included survivin in this study based on the promising results we had observed with this antigen, in terms of elevated expression, in B‐ALL compared with HV samples previously (Boullosa et al , ). All cell lines expressed all housekeeping genes as well as the four antigens under investigation, except BMX which was not expressed in any of the cells lines.…”

New targets for therapy: antigen identification in adults with B-cell acute lymphoblastic leukaemia

“…The final group incorporates viral antigens and includes the protein products of human papilloma virus (ie E6 and E7 early antigens) and Epstein-Barr virus (ie EBNA). To date, the techniques we have used to identify antigens have predominantly found overexpressed antigens such as HAGE [20], PASD1 [21], SSX2IP [17,18] and survivin [22]. Although the reason for the overexpression has not been characterised by our group, it is likely that global promoter hypomethylation, seen in many cancers, and a demonstrated cause of CTA overexpression, belies this finding.…”

“…We have used several techniques to identify antigens in AML, colon cancer, ovarian cancer and B-cell ALL. These techniques examine patient samples through the analysis of RNA using RT-PCR [20], qPCR [22,29] and expression microarrays [30,31], urine [32], or peptides presented on MHC, using mass spectrometry [33], antibodies, i.e. serological analysis of recombinant cDNA expression libraries (SEREX) [21,24] and protein microarrays [34] and we have prioritised epitope specific T-cell responses using peptide-MHC arrays [27].…”