Beta-2-microglobulin gene transfer in hla class i deficient tumor cells using recombinant adenovirus
- Autores: Ana Belén del Campo Alonso
- Directores de la Tesis: Federico Garrido Torres-Puchol (dir. tes.), Natalia Aptsiauri (codir. tes.), Gloria González Aseguinolaza (codir. tes.)
- Lectura: En la Universidad de Granada ( España ) en 2014
- Idioma: español
- Tribunal Calificador de la Tesis: Enrique García Olivares (presid.), María Teresa Cabrera Castillo (secret.), Annette Paschen (voc.), Francisco Martín (voc.), Rubén Hernández-Alcoceba (voc.)
- Materias:
- Enlaces
- Tesis en acceso abierto en: DIGIBUG
- Resumen
Antitumor immune response and the success of cancer immunotherapy depend on the proper recognition of the tumor HLA class I complex (HLA class I heavy chain/ ß2-microglobulin/peptide) by cytotoxic T-cells. It has become apparent that lack of tumor rejection is the result of immune selection and escape of tumor cells that develop low immunogenic phenotype. Altered expression of tumor HLA class I molecules is a frequent event described in almost all types of cancer and often is associated with metastatic dissemination, poor prognosis and resistance to immunotherapy. These defects can occur at the genetic, epigenetic, transcriptional and posttranscriptional levels and represent either regulatory abnormalities, which can be recovered with cytokine treatment, or structural alterations, including genetic/chromosomal defects. However, the structural defects may have more serious implications on T-cell-mediated tumor rejection and, ultimately, on the outcome of cancer immunotherapy.
In this study we provide evidence supporting the accumulation of HLA class I loss in metastatic melanoma and immune escape of HLA-negative tumor cells. We have been able to illustrate the immune escape of HLA class I negative tumor cells and chronological sequence of appearance of ß2m gene mutation in successive lesions obtained from a patient with metastatic melanoma resistant to DC vaccination. We detected in the ß2m-negative nodules an early onset of ß2m loss due to a combination of a novel mutation at codon 67 of exon 2 of the ß2m gene, and loss of the second allele by LOH at chromosome 15. It is likely that tumor cells with HLA class I alterations in this patient first escaped from the immune attack during natural metastatic progression and that HLA-negative immune escape variants were further immunoselected during the vaccination, which might explain the failure of the therapy as the gradual decrease in HLA expression correlated with decrease T-cell infiltration (CD8+).
Thus, strategies to overcome the lack of HLA class I expression require consideration and targeting of ß2m as an attractive option to recover HLA class I expression in ¿2m-negative tumor cells in patients with metastatic progression or recurrent tumors. For this purpose, we have constructed a replication-deficient adenoviral vector carrying human ß2m gene (AdCMVß2m) and characterized its efficacy to recover HLA class I expression using different human cancer cell lines with structural defects in ß2m. We show that in vitro transduced tumor cells become sensitive to lysis by peptide-stimulated HLA-restricted T-cells, and recover the ability to induce peptide-specific IFN¿¿secretion by T cells in a HLA-restricted manner without compromising the antigen processing and presentation. In in vivo experiments using human tumour xenograft model, the intratumoral injection of AdCMVß2m also led to restoration of normal HLA class I expression.
Our data support the clinical application of such vector in patients with metastatic cancer with structural defects in ß2m gene and/or LOH in chromosome 15, as in such patients, the immunotherapy might lead to generation of metastatic lesions with irreversible defects in HLA class I expression that would not respond to therapy and eventually would progress. Our findings emphasize the importance of carefully defining the molecular mechanisms responsible for a particular altered HLA class I phenotype to design specific ways to restore in situ normal tumor HLA class I expression. Therefore, the optimization of the existing immunotherapy approaches will greatly benefit from the characterization of the HLA class I alterations in primary tumors and from the analysis of the correlation of impaired HLA expression with metastatic progression.
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