Alternative BCR/ABL Splice Variants in Philadelphia Chromosome-Positive Leukemias Result in Novel Tumor-Specific Fusion Proteins that May Represent Potential Targets for Immunotherapy Approaches

Gisella Volpe, Alessandro Cignetti, Cristina Panuzzo, Mirela Kuka, Katiuscia Vitaggio, Mara Brancaccio, Giuseppe Perrone, Monica Rinaldi, Giuseppina Prato, Milena Fava, Massimo Geuna, Marisa Pautasso, Claudia Casnici, Emanuela Signori, Giancarlo Tonon, Guido Tarone, Ornella Marelli, Vito M. Fazio, Giuseppe Saglio

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Imatinib currently represents the standard treatment in the early chronic phase of chronic myelogenous leukemia (CML), thanks to the high percentage of cytogenetic complete remission achieved, but it is yet unclear to what extent it can eradicate leukemia. Therefore, different vaccination strategies have been suggested, mainly based on the exploitment of the junctional peptides spanning the fusion region of the Bcr/Abl proteins. To identify new potential immunologic targets, 63 Philadelphia chromosome-positive patients and 6 BCR/ABL-positive cell lines were tested in nested reverse transcriptase PCR to detect the presence of BCR/ABL transcripts arising from the alternative splicing of the main BCR/ABL transcripts. We could detect BCR/ABL transcripts with junctions between BCR exon 1, 13, or 14 and ABL exon 4 in ∼80% of patients and 84% of cell lines, beside the main fusion transcripts. Translation products of these transcripts were characterized at their COOH terminus by a large amino acid portion derived from the out of frame (OOF) reading of ABL gene. These proteins were detected in BCR/ABL-positive cell lines by immunoprecipitation and immunohistochemistry. Finally, we determined whether OOF-specific CD8 + T cells could be found in the peripheral blood of CML patients and whether they could acquire effector function following in vitro sensitization with OOF-derived peptides predicted to bind to human leucocyte antigen (HLA)-A2 and HLA-A3 molecules. We detected the presence of OOF-specific CD8+ Tcells in four of four patients studied, and in one case, these Tc ells exhibited specific cytotoxic activity against both peptide-pulsed targets and autologous primary CML cells.

Original languageEnglish
Pages (from-to)5300-5307
Number of pages8
JournalCancer Research
Volume67
Issue number11
DOIs
Publication statusPublished - Jun 1 2007

Fingerprint

Philadelphia Chromosome
Immunotherapy
Leukemia
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
HLA Antigens
Cell Line
Peptides
Exons
Neoplasms
Proteins
bcr-abl Fusion Proteins
Leukemia, Myeloid, Chronic Phase
Reading Frames
Alternative Splicing
Reverse Transcriptase Polymerase Chain Reaction
Immunoprecipitation
varespladib methyl
Cytogenetics
Vaccination
Immunohistochemistry

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Alternative BCR/ABL Splice Variants in Philadelphia Chromosome-Positive Leukemias Result in Novel Tumor-Specific Fusion Proteins that May Represent Potential Targets for Immunotherapy Approaches. / Volpe, Gisella; Cignetti, Alessandro; Panuzzo, Cristina; Kuka, Mirela; Vitaggio, Katiuscia; Brancaccio, Mara; Perrone, Giuseppe; Rinaldi, Monica; Prato, Giuseppina; Fava, Milena; Geuna, Massimo; Pautasso, Marisa; Casnici, Claudia; Signori, Emanuela; Tonon, Giancarlo; Tarone, Guido; Marelli, Ornella; Fazio, Vito M.; Saglio, Giuseppe.

In: Cancer Research, Vol. 67, No. 11, 01.06.2007, p. 5300-5307.

Research output: Contribution to journalArticle

Volpe, G, Cignetti, A, Panuzzo, C, Kuka, M, Vitaggio, K, Brancaccio, M, Perrone, G, Rinaldi, M, Prato, G, Fava, M, Geuna, M, Pautasso, M, Casnici, C, Signori, E, Tonon, G, Tarone, G, Marelli, O, Fazio, VM & Saglio, G 2007, 'Alternative BCR/ABL Splice Variants in Philadelphia Chromosome-Positive Leukemias Result in Novel Tumor-Specific Fusion Proteins that May Represent Potential Targets for Immunotherapy Approaches', Cancer Research, vol. 67, no. 11, pp. 5300-5307. https://doi.org/10.1158/0008-5472.CAN-06-3737
Volpe, Gisella ; Cignetti, Alessandro ; Panuzzo, Cristina ; Kuka, Mirela ; Vitaggio, Katiuscia ; Brancaccio, Mara ; Perrone, Giuseppe ; Rinaldi, Monica ; Prato, Giuseppina ; Fava, Milena ; Geuna, Massimo ; Pautasso, Marisa ; Casnici, Claudia ; Signori, Emanuela ; Tonon, Giancarlo ; Tarone, Guido ; Marelli, Ornella ; Fazio, Vito M. ; Saglio, Giuseppe. / Alternative BCR/ABL Splice Variants in Philadelphia Chromosome-Positive Leukemias Result in Novel Tumor-Specific Fusion Proteins that May Represent Potential Targets for Immunotherapy Approaches. In: Cancer Research. 2007 ; Vol. 67, No. 11. pp. 5300-5307.
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abstract = "Imatinib currently represents the standard treatment in the early chronic phase of chronic myelogenous leukemia (CML), thanks to the high percentage of cytogenetic complete remission achieved, but it is yet unclear to what extent it can eradicate leukemia. Therefore, different vaccination strategies have been suggested, mainly based on the exploitment of the junctional peptides spanning the fusion region of the Bcr/Abl proteins. To identify new potential immunologic targets, 63 Philadelphia chromosome-positive patients and 6 BCR/ABL-positive cell lines were tested in nested reverse transcriptase PCR to detect the presence of BCR/ABL transcripts arising from the alternative splicing of the main BCR/ABL transcripts. We could detect BCR/ABL transcripts with junctions between BCR exon 1, 13, or 14 and ABL exon 4 in ∼80{\%} of patients and 84{\%} of cell lines, beside the main fusion transcripts. Translation products of these transcripts were characterized at their COOH terminus by a large amino acid portion derived from the out of frame (OOF) reading of ABL gene. These proteins were detected in BCR/ABL-positive cell lines by immunoprecipitation and immunohistochemistry. Finally, we determined whether OOF-specific CD8 + T cells could be found in the peripheral blood of CML patients and whether they could acquire effector function following in vitro sensitization with OOF-derived peptides predicted to bind to human leucocyte antigen (HLA)-A2 and HLA-A3 molecules. We detected the presence of OOF-specific CD8+ Tcells in four of four patients studied, and in one case, these Tc ells exhibited specific cytotoxic activity against both peptide-pulsed targets and autologous primary CML cells.",
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AU - Panuzzo, Cristina

AU - Kuka, Mirela

AU - Vitaggio, Katiuscia

AU - Brancaccio, Mara

AU - Perrone, Giuseppe

AU - Rinaldi, Monica

AU - Prato, Giuseppina

AU - Fava, Milena

AU - Geuna, Massimo

AU - Pautasso, Marisa

AU - Casnici, Claudia

AU - Signori, Emanuela

AU - Tonon, Giancarlo

AU - Tarone, Guido

AU - Marelli, Ornella

AU - Fazio, Vito M.

AU - Saglio, Giuseppe

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