Reul and Frisch et al. used a HER2-targeted adeno-associated viral (AAV) vector to deliver genes encoding immune checkpoint inhibitors to tumors. In HER2+ tumor-bearing mice, i.v. treatment with either HER2-AAV or untargeted AAV induced anti-PD-1 expression in the tumor, but HER2-AAV caused less accumulation of anti-PD-1 in the liver and serum. When combined with chemotherapy, HER2-AAV-anti-PD-1 marginally reduced tumor progression. The researchers also tested HER2-AAV delivery of genes for human nivolumab, which targeted tumors, but nivolumab was expressed at low levels compared to the murine anti-PD-1 construct.
Immune checkpoint inhibitors (ICIs) can block distinct receptors on T cells or tumor cells thus preventing T cell inactivation and tumor immune escape. While the clinical response to treatment with ICIs in cancer patients is impressive, this therapy is often associated with a number of immune-related adverse events. There is therefore a need to explore innovative strategies of tumor-specific delivery of ICIs. Delivery of therapeutic proteins on a genetic level can be accomplished with viral vectors including those derived from adeno-associated virus (AAV). Here, we assessed the tumor-targeted Her2-AAV, a receptor-targeted AAV vector binding to the tumor antigen Her2/neu for cell entry, as vehicle for ICI gene delivery. Initially, we packaged the coding sequence of a scFv-Fc fusion protein directed against mouse programmed cell death protein-1 (PD-1) into Her2-AAV. Upon transduction of Her2/neu(+) RENCA cells, AAV-encoded alphaPD-1 was readily detectable in the cell culture supernatant and revealed specific binding to its target antigen. In vivo, in BALB/c mice bearing subcutaneous RENCA-Her2/neu tumors, Her2-AAV mediated specific gene delivery into tumor tissue upon intravenous administration as verified by luciferase gene transfer and in vivo imaging thus demonstrating unimpaired tumor-targeting by Her2-AAV vectors in immunocompetent animals. When delivering the alphaPD-1 gene, levels of ICI were similar in tumor tissue for Her2-AAV and AAV2 but substantially reduced in liver for Her2-AAV. When combined with chemotherapy a tendency for reduced progression of tumor growth was documented for Her2-AAV treated mice. To get closer to the clinical situation, AAV constructs that deliver the complete coding sequence of the therapeutic antibody nivolumab which is directed against human PD-1 were generated next. The AAV-Nivolumab constructs were expressed and released from transduced MDA-MB-453 cells in vitro and from RENCA-Her2/neu cells upon intratumoral as well as intravenous administration in vivo. Antibody processing and expression levels were further improved through optimization of construct design. In conclusion, we provide proof-of-principle for redirecting the biodistribution of ICIs from liver and serum to tumor tissue by the use of engineered AAV vectors. This strategy can be easily combined with other types of immunotherapeutic concepts.
Author Info: (1) Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, Langen, Germany. German Cancer Consortium (DKTK), Heidelberg, Germany. (2) Molecular Biotechnology and Gene The
Author Info: (1) Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, Langen, Germany. German Cancer Consortium (DKTK), Heidelberg, Germany. (2) Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, Langen, Germany. (3) National Center for Tumor Diseases, Heidelberg, Germany. (4) Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, Langen, Germany. (5) Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, Langen, Germany. (6) National Center for Tumor Diseases, Heidelberg, Germany. (7) Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, Langen, Germany. German Cancer Consortium (DKTK), Heidelberg, Germany.
