Scheuermann, S. | Search bioRxiv

bioRxiv2026

Pan-B-Lineage Targeting with Adapter CAR-T Cells Controls Antigen-Heterogeneous Lymphoma

Antigen heterogeneity and antigen-negative relapse represent major limitations to durable CAR-T cell efficacy in B-lineage malignancies. We previously developed the Adapter CAR-T cell (AdCAR-T) platform, which enables flexible redirection of engineered T cells to distinct surface antigens through biotinylated adapter molecules (AMs). In this study, AMs generated from an in-house-produced tafasitamab biosimilar (anti-CD19), commercial rituximab (anti-CD20), and an in-house-produced daratumumab biosimilar (anti-CD38) mediated potent, antigen-specific AdCAR-T cell cytotoxicity. While single-antigen targeting resulted in the selection of antigen-negative tumor populations, simultaneous targeting of CD19, CD20, and CD38 effectively controlled a defined heterogeneous Burkitt lymphoma model in vitro and induced sustained tumor control in vivo. Selective loss of the CD38+ AdCAR-T cell population after CD38-directed AM exposure was consistent with fratricide; however, the surviving CD38low population retained cytotoxic activity. These findings establish combinatorial AdCAR-T cell targeting as a flexible pan-B-lineage strategy for addressing pre-existing antigen heterogeneity and support further development of antibody-derived AM combinations for B-cell malignancies.

immunology↗

bioRxiv2021

TissueGrinder, a novel technology for rapid generation of patient-derived single cell suspensions from solid tumors by mechanical tissue dissociation

IntroductionRecent advances hold promise of making personalized medicine a step closer to implementation in clinical settings. However, traditional sample preparation methods are not robust and reproducible. In this study, the TissueGrinder, a novel mechanical semi-automated benchtop device, which can isolate cells from tissue in a very fast and enzyme-free way is tested for cell isolation from surgically resected tumor tissues. Methods33 surgically resected tumor tissues from various but mainly pancreatic, liver or colorectal origins were processed by both novel TissueGrinder and explant method. An optimized processing program for tumors from pancreatic, liver or colorectal cancer was developed. The viability and morphological characteristics of the isolated cells were evaluated microscopically. Expression of pancreatic cancer markers was evaluated in cells isolated from pancreatic tumors. Finally, the effect of mechanical stress on the cells was evaluated by assessing apoptosis markers via western blotting. ResultsTissueGinder was more efficient in isolating cells from tumor tissue with a success rate of 75% when compared to explant method 45% in terms of cell outgrowth six weeks after processing. Cells isolated with TissueGinder had a higher abundance and were more heterogeneous in composition as compared to explant method. Mechanical processing of the cells with TissueGrinder does not lead to apoptosis but causes slight stress to the cells. DiscussionOur results show that TissueGrinder can process solid tumor tissues more rapidly and efficiently and with higher success rate compared to the conventionally used explant method. The results of the study suggest that the TissueGrinder might be a suitable method for obtaining cells, which is important for its application in individualized therapy. Due to the great variance in different tumor entities and the associated individual tissue characteristics, a further development of the dissociation protocol for other types of tumors and normal will be targeted.

cancer biology↗