Biology subjects

Blankenstein, T.

Publications and source records attributed to Blankenstein, T..

3 recordsLinked to original sources

bioRxiv2024

Oncogene inactivation-induced senescence facilitates tumor relapse

Oncogene-directed therapies can induce profound tumor regression in oncogene-addicted cancers such as BRAF-mutant melanoma and KRAS-driven pancreatic cancer, but their long-term benefit is often limited by resistance and early relapse. The mechanisms that allow residual cells to adapt, persist in a dormant state, and eventually fuel recurrence remain poorly understood. Here, we show that oncogene inactivation rapidly induces hallmark features of senescence together with a pro-inflammatory senescence-associated secretory phenotype (SASP). In vivo, oncogene inactivation-induced senescence (OIIS) predisposed tumors to relapse, characterized by polyploidy, chromosomal instability, and acquisition of alternative oncogenic pathways such as Mdm2 upregulation. Tumor microenvironment profiling by spectral flow cytometry revealed that relapse was associated with neovascularization and a shift from an immune-activated to an immunosuppressive milieu, indicating that senescent cells remodel their niche to promote regrowth. Importantly, OIIS features were also observed in human BRAFV600E melanoma cells treated with vemurafenib, confirming the clinical relevance of our findings. Together, our findings establish OIIS as a double-edged process: it initially restrains tumor growth but simultaneously creates conditions that favor recurrence. By defining the genetic, metabolic, cytogenetic, and microenvironmental hallmarks of OIIS, our study highlights adaptations to oncogene deprivation that limit the durability of targeted therapies.

cancer biology↗

bioRxiv2024

Targeting MYCN upregulates L1CAM tumor antigen in MYCN-dysregulated neuroblastoma to increase CAR T cell efficacy

BackgroundCurrent treatment protocols have only limited success in pediatric patients with neuroblastomas harboring amplifications of the central oncogene, MYCN. Adoptive T cell therapy presents an innovative strategy to improve cure rates. However, L1CAM-targeting CAR T cells achieved only limited response against refractory/relapsed neuroblastoma in an ongoing phase I trial to date. Here, we investigate how oncogenic MYCN levels influence tumor cell response to CAR T cells, as one possible factor limiting success in trials. MethodsHigh MYCN levels were induced in SK-N-AS cells harboring the normal diploid MYCN complement using a tetracycline-inducible system. The inducible MYCN cell model or MYCN-amplified neuroblastoma cell lines were cocultured with L1CAM-CAR T cells. CAR T cell effector function was assessed via activation marker expression (flow cytometry), cytokine release and tumor cytotoxicity (biophotonic signal assessment). The cell model was characterized using RNA sequencing, and our data compared to publicly available RNA and proteomic data sets from neuroblastomas. ChIP-sequencing data was used to determine transcriptional L1CAM regulation by MYCN using public data sets. Synergism between CAR T cells and the MLN8237 AURKA inhibitor, which indirectly inhibits MYCN activity, was assessed in vitro using the Bliss model and in vivo in an immunocompromised mouse model. ResultsInducing high MYCN levels in the neuroblastoma cell model reduced L1CAM expression and, consequently, L1CAM-CAR T cell effector function (activation, cytokine release and cytotoxicity) in vitro. Primary neuroblastomas possessing high MYCN levels expressed lower levels of both the L1CAM transcript and L1CAM tumor antigen. Indirectly inhibiting MYCN via AURKA using MLN8237 treatment restored L1CAM expression on tumor cells in vitro and restored L1CAM-CAR T cell effector function. Combining MLN8237 and L1CAM-CAR T cell treatment synergistically increased neuroblastoma-directed killing in MYCN-overexpressing cells in vitro and in vivo concomitant with severe in vivo toxicity. ConclusionWe shed new light on a primary resistance mechanism in MYCN-driven neuroblastoma against L1CAM-CAR T cells via target antigen downregulation. These data suggest that combining L1CAM-CAR T cell therapy with pharmacological MYCN inhibition may benefit patients with high-risk neuroblastomas harboring MYCN amplifications.

immunology↗

bioRxiv2018

Identification and Ranking of Recurrent Neo-Epitopes in Cancer

Neo-epitopes are emerging as attractive targets for cancer immunotherapy and new strategies for rapid identification of relevant candidates have become a priority. We propose a method for in silico selection of candidates which have a high potential for neo-antigen generation and are likely to appear in multiple patients. This is achieved by carefully screening 33 TCGA data sets for recurrent somatic amino acid exchanges and, for the 1,055 resulting recurrent variants, applying MHC class I binding prediction algorithms. A preliminary confirmation of epitope binding and recognition by CD8 T cells has been carried out for a couple of candidates in humanized mice. Recurrent neo-epitopes may be suitable to supplement existing personalized T cell treatment approaches with precision treatment options.

genomics↗